{"id":9405,"date":"2023-08-11T09:10:36","date_gmt":"2023-08-11T09:10:36","guid":{"rendered":"https:\/\/vospan.com\/?p=9405"},"modified":"2024-03-20T08:15:45","modified_gmt":"2024-03-20T08:15:45","slug":"cong-nghe-tien-tien-trong-kinh-hien-vi-quan-sat-te-bao-song","status":"publish","type":"post","link":"https:\/\/vospan.com\/cong-nghe-tien-tien-trong-kinh-hien-vi-quan-sat-te-bao-song\/","title":{"rendered":"C\u00f4ng ngh\u1ec7 ti\u00ean ti\u1ebfn trong k\u00ednh hi\u1ec3n vi quan s\u00e1t t\u1ebf b\u00e0o s\u1ed1ng"},"content":{"rendered":"<div id=\"ez-toc-container\" class=\"ez-toc-v2_0_68_1 counter-hierarchy ez-toc-counter ez-toc-grey ez-toc-container-direction\">\n<div class=\"ez-toc-title-container\">\n<label class=\"ez-toc-title ez-toc-toggle\" style=\"cursor: pointer\">T&oacute;m t\u1eaft n\u1ed9i dung<\/label>\n<span class=\"ez-toc-title-toggle\"><a href=\"#\" class=\"ez-toc-pull-right ez-toc-btn ez-toc-btn-xs ez-toc-btn-default ez-toc-toggle\" aria-label=\"Toggle Table of Content\"><span class=\"ez-toc-js-icon-con\"><span class=\"\"><span class=\"eztoc-hide\" style=\"display:none;\">Toggle<\/span><span class=\"ez-toc-icon-toggle-span\"><svg style=\"fill: #0a0a0a;color:#0a0a0a\" xmlns=\"http:\/\/www.w3.org\/2000\/svg\" class=\"list-377408\" width=\"20px\" height=\"20px\" viewBox=\"0 0 24 24\" fill=\"none\"><path d=\"M6 6H4v2h2V6zm14 0H8v2h12V6zM4 11h2v2H4v-2zm16 0H8v2h12v-2zM4 16h2v2H4v-2zm16 0H8v2h12v-2z\" fill=\"currentColor\"><\/path><\/svg><svg style=\"fill: #0a0a0a;color:#0a0a0a\" class=\"arrow-unsorted-368013\" xmlns=\"http:\/\/www.w3.org\/2000\/svg\" width=\"10px\" height=\"10px\" viewBox=\"0 0 24 24\" version=\"1.2\" baseProfile=\"tiny\"><path d=\"M18.2 9.3l-6.2-6.3-6.2 6.3c-.2.2-.3.4-.3.7s.1.5.3.7c.2.2.4.3.7.3h11c.3 0 .5-.1.7-.3.2-.2.3-.5.3-.7s-.1-.5-.3-.7zM5.8 14.7l6.2 6.3 6.2-6.3c.2-.2.3-.5.3-.7s-.1-.5-.3-.7c-.2-.2-.4-.3-.7-.3h-11c-.3 0-.5.1-.7.3-.2.2-.3.5-.3.7s.1.5.3.7z\"\/><\/svg><\/span><\/span><\/span><\/a><\/span><\/div>\n<nav><ul class='ez-toc-list ez-toc-list-level-1 eztoc-toggle-hide-by-default' ><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-1\" href=\"https:\/\/vospan.com\/cong-nghe-tien-tien-trong-kinh-hien-vi-quan-sat-te-bao-song\/#Gioi_thieu\" title=\"Gi\u1edbi thi\u1ec7u\">Gi\u1edbi thi\u1ec7u<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-2\" href=\"https:\/\/vospan.com\/cong-nghe-tien-tien-trong-kinh-hien-vi-quan-sat-te-bao-song\/#Kinh_hien_vi_huynh_quang_va_ky_thuat_truyen_sang\" title=\"K\u00ednh hi\u1ec3n vi hu\u1ef3nh quang v\u00e0 k\u1ef9 thu\u1eadt truy\u1ec1n s\u00e1ng\">K\u00ednh hi\u1ec3n vi hu\u1ef3nh quang v\u00e0 k\u1ef9 thu\u1eadt truy\u1ec1n s\u00e1ng<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-3\" href=\"https:\/\/vospan.com\/cong-nghe-tien-tien-trong-kinh-hien-vi-quan-sat-te-bao-song\/#Kinh_hien_vi_huynh_quang_truong_rong\" title=\"K\u00ednh hi\u1ec3n vi hu\u1ef3nh quang tr\u01b0\u1eddng r\u1ed9ng\">K\u00ednh hi\u1ec3n vi hu\u1ef3nh quang tr\u01b0\u1eddng r\u1ed9ng<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-4\" href=\"https:\/\/vospan.com\/cong-nghe-tien-tien-trong-kinh-hien-vi-quan-sat-te-bao-song\/#Kinh_hien_vi_dong_tieu_quet_laze\" title=\"K\u00ednh hi\u1ec3n vi \u0111\u1ed3ng ti\u00eau qu\u00e9t laze\">K\u00ednh hi\u1ec3n vi \u0111\u1ed3ng ti\u00eau qu\u00e9t laze<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-5\" href=\"https:\/\/vospan.com\/cong-nghe-tien-tien-trong-kinh-hien-vi-quan-sat-te-bao-song\/#Kinh_hien_vi_dong_tieu_dia_quay\" title=\"K\u00ednh hi\u1ec3n vi \u0111\u1ed3ng ti\u00eau \u0111\u0129a quay\">K\u00ednh hi\u1ec3n vi \u0111\u1ed3ng ti\u00eau \u0111\u0129a quay<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-6\" href=\"https:\/\/vospan.com\/cong-nghe-tien-tien-trong-kinh-hien-vi-quan-sat-te-bao-song\/#Kinh_hien_vi_da_diem\" title=\"K\u00ednh hi\u1ec3n vi \u0111a \u0111i\u1ec3m\">K\u00ednh hi\u1ec3n vi \u0111a \u0111i\u1ec3m<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-7\" href=\"https:\/\/vospan.com\/cong-nghe-tien-tien-trong-kinh-hien-vi-quan-sat-te-bao-song\/#Ung_dung_kinh_hien_vi_huynh_quang\" title=\"\u1ee8ng d\u1ee5ng k\u00ednh hi\u1ec3n vi hu\u1ef3nh quang\u00a0\">\u1ee8ng d\u1ee5ng k\u00ednh hi\u1ec3n vi hu\u1ef3nh quang\u00a0<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-8\" href=\"https:\/\/vospan.com\/cong-nghe-tien-tien-trong-kinh-hien-vi-quan-sat-te-bao-song\/#Ket_luan\" title=\"K\u1ebft lu\u1eadn\">K\u1ebft lu\u1eadn<\/a><\/li><\/ul><\/nav><\/div>\n<h2><span class=\"ez-toc-section\" id=\"Gioi_thieu\"><\/span><strong>Gi\u1edbi thi\u1ec7u<\/strong><span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p>Theo d\u00f5i h\u00ecnh \u1ea3nh chuy\u1ec3n \u0111\u1ed9ng c\u1ee7a t\u1ebf b\u00e0o s\u1ed1ng theo ph\u01b0\u01a1ng ph\u00e1p truy\u1ec1n th\u1ed1ng b\u1eb1ng c\u00e1ch s\u1eed d\u1ee5ng c\u00e1c k\u1ef9 thu\u1eadt t\u0103ng c\u01b0\u1eddng \u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n ph\u1ed5 bi\u1ebfn bao g\u1ed3m tr\u01b0\u1eddng s\u00e1ng, \u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n nhi\u1ec5u vi sai (DIC), \u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n \u0111i\u1ec1u bi\u1ebfn Hoffman <b>(<\/b>HMC<b>)<\/b>, t\u01b0\u01a1ng ph\u1ea3n pha v\u00e0 hu\u1ef3nh quang tr\u01b0\u1eddng r\u1ed9ng. Tuy nhi\u00ean, c\u00e1c k\u1ef9 thu\u1eadt m\u1edbi hi\u1ec7n nay kh\u00f4ng ch\u1ec9 \u0111\u01a1n gi\u1ea3n l\u00e0 t\u1ea1o ra chu\u1ed7i c\u00e1c \u1ea3nh v\u1ec1 c\u1ea5u tr\u00fac v\u00e0 ch\u1ee9c n\u0103ng c\u1ee7a t\u1ebf b\u00e0o, m\u00e0 c\u00f2n cho ph\u00e9p tua nhanh h\u00ecnh \u1ea3nh \u0111\u01b0\u1ee3c t\u00edch h\u1ee3p v\u1edbi c\u00e1c ch\u1ebf \u0111\u1ed9 chuy\u00ean bi\u1ec7t \u0111\u1ec3 theo d\u00f5i, \u0111o l\u01b0\u1eddng v\u00e0 g\u00e2y nhi\u1ec5u c\u00e1c ho\u1ea1t \u0111\u1ed9ng \u0111\u1ed9ng c\u1ee7a m\u00f4, t\u1ebf b\u00e0o, v\u00e0 c\u00e1c c\u1ea5u tr\u00fac d\u01b0\u1edbi t\u1ebf b\u00e0o.<\/p>\n<p>Ph\u1ea7n l\u1edbn c\u00e1c cu\u1ed9c nghi\u00ean c\u1ee9u h\u00ecnh \u1ea3nh t\u1ebf b\u00e0o s\u1ed1ng \u0111\u01b0\u1ee3c th\u1ef1c hi\u1ec7n v\u1edbi c\u00e1c t\u1ebf b\u00e0o \u0111\u1ed9ng v\u1eadt c\u00f3 v\u00fa c\u00f3 \u0111\u1ed9 d\u00e0y t\u1eeb 10 micromet. Tuy nhi\u00ean, c\u00e1c nh\u00e0 nghi\u00ean c\u1ee9u \u0111ang ng\u00e0y c\u00e0ng ch\u00fa \u00fd \u0111\u1ebfn c\u00e1c m\u1eabu m\u00f4 \u0111\u1ed9ng v\u1eadt v\u00e0 th\u1ef1c v\u1eadt d\u00e0y h\u01a1n c\u00f3 th\u1ec3 c\u00f3 \u0111\u1ed9 d\u00e0y t\u1eeb 10 \u0111\u1ebfn 200 micromet. Trong h\u1ea7u nh\u01b0 t\u1ea5t c\u1ea3 c\u00e1c t\u00ecnh hu\u1ed1ng ch\u1ee5p \u1ea3nh t\u1ebf b\u00e0o s\u1ed1ng s\u1eed d\u1ee5ng <a href=\"https:\/\/vospan.com\/product-brands\/zeiss-vi\/\">k\u00ednh hi\u1ec3n vi<\/a> tr\u01b0\u1eddng r\u1ed9ng, th\u00f4ng tin ngo\u00e0i ti\u00eau c\u1ef1 l\u00e0m m\u1edd h\u00ecnh \u1ea3nh v\u00e0 s\u1ef1 khu\u1ea5y tr\u1ed9n li\u00ean t\u1ee5c c\u1ee7a t\u1ebf b\u00e0o ch\u1ea5t t\u1ea1o ra nh\u1eefng h\u1ea1n ch\u1ebf v\u1ec1 th\u1eddi gian ph\u01a1i s\u00e1ng. C\u1ea3 hai ph\u01b0\u01a1ng ph\u00e1p tr\u01b0\u1eddng s\u00e1ng v\u00e0 hu\u1ef3nh quang \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng \u0111\u1ec3 ch\u1ee5p \u1ea3nh c\u00e1c m\u00f4 \u0111\u1ed9ng v\u1eadt v\u00e0 th\u1ef1c v\u1eadt m\u1ecfng c\u0169ng nh\u01b0 d\u00e0y h\u01a1n \u0111\u1ec1u ph\u1ea3i t\u00ednh \u0111\u1ebfn \u0111\u1ed9 nh\u1ea1y c\u1ee7a c\u00e1c m\u1eabu v\u1eadt n\u00e0y khi ti\u1ebfp x\u00fac v\u1edbi \u00e1nh s\u00e1ng v\u00e0 c\u00e1c v\u1ea5n \u0111\u1ec1 li\u00ean quan \u0111\u1ebfn vi\u1ec7c gi\u1ea3i quy\u1ebft c\u00e1c \u0111\u1eb7c \u0111i\u1ec3m n\u1eb1m tr\u00ean 20 \u0111\u1ebfn 30 micromet trong m\u1eabu v\u1eadt.<\/p>\n<p><img decoding=\"async\" class=\"aligncenter\" src=\"https:\/\/zeiss-campus.magnet.fsu.edu\/articles\/livecellimaging\/images\/techniquesfigure1.jpg\" \/><\/p>\n<p>C\u00e1c k\u1ef9 thu\u1eadt tr\u01b0\u1eddng s\u00e1ng th\u01b0\u1eddng \u00edt g\u00e2y h\u1ea1i h\u01a1n cho c\u00e1c t\u1ebf b\u00e0o s\u1ed1ng, nh\u01b0ng ph\u01b0\u01a1ng ph\u00e1p quan s\u00e1t c\u00e1c protein c\u1ee5 th\u1ec3 b\u1eb1ng c\u00e1ch s\u1eed d\u1ee5ng \u00e1nh s\u00e1ng truy\u1ec1n qua ch\u01b0a \u0111\u01b0\u1ee3c ph\u00e1t tri\u1ec3n r\u1ed9ng r\u00e3i. Vi\u1ec7c t\u1ea1o ra s\u1ef1 kh\u00e1c bi\u1ec7t v\u1ec1 c\u01b0\u1eddng \u0111\u1ed9 ho\u1eb7c m\u00e0u s\u1eafc c\u00f3 \u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n cao trong h\u00ecnh \u1ea3nh tr\u01b0\u1eddng s\u00e1ng kh\u00f3 h\u01a1n vi\u1ec7c x\u00e1c \u0111\u1ecbnh s\u1ef1 thay \u0111\u1ed5i c\u01b0\u1eddng \u0111\u1ed9 s\u00e1ng (th\u1ef1c ch\u1ea5t l\u00e0 do hu\u1ef3nh quang) tr\u00ean n\u1ec1n t\u1ed1i ho\u1eb7c \u0111en. Do \u0111\u00f3, c\u00e1c k\u1ef9 thu\u1eadt tr\u01b0\u1eddng s\u00e1ng t\u00ecm th\u1ea5y c\u00e1c \u1ee9ng d\u1ee5ng trong vi\u1ec7c theo d\u00f5i c\u00e1c b\u00e0o quan ho\u1eb7c ph\u00e1t hi\u1ec7n tr\u00ean to\u00e0n t\u1ebf b\u00e0o, trong khi c\u00e1c ph\u01b0\u01a1ng ph\u00e1p hu\u1ef3nh quang, bao g\u1ed3m c\u00e1c k\u1ef9 thu\u1eadt \u0111\u1ed3ng ti\u00eau, th\u01b0\u1eddng \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng \u0111\u1ec3 theo d\u00f5i c\u00e1c ph\u00e2n t\u1eed c\u1ee5 th\u1ec3.<\/p>\n<p>Tr\u00ecnh b\u00e0y trong <em>H\u00ecnh 1<\/em> l\u00e0 m\u1ed9t s\u01a1 \u0111\u1ed3 minh h\u1ecda c\u00e1c ch\u1ebf \u0111\u1ed9 h\u00ecnh \u1ea3nh ph\u1ed5 bi\u1ebfn trong ch\u1ebf \u0111\u1ed9 qu\u00e9t v\u00e0 tr\u01b0\u1eddng r\u1ed9ng c\u1ee7a k\u00ednh hi\u1ec3n vi hu\u1ef3nh quang. C\u00e1c k\u1ef9 thu\u1eadt tr\u01b0\u1eddng r\u1ed9ng, qu\u00e9t laser, \u0111\u0129a quay v\u00e0 \u0111a \u0111i\u1ec3m s\u1eed d\u1ee5ng c\u00e1c chi\u1ebfn l\u01b0\u1ee3c ph\u00e1t hi\u1ec7n v\u00e0 chi\u1ebfu s\u00e1ng r\u1ea5t kh\u00e1c nhau \u0111\u1ec3 t\u1ea1o \u1ea3nh. S\u01a1 \u0111\u1ed3 minh h\u1ecda m\u1ed9t t\u1ebf b\u00e0o \u0111\u1ed9ng v\u1eadt c\u00f3 v\u00fa b\u00e1m d\u00ednh tr\u00ean m\u1ed9t l\u1edbp ph\u1ee7 \u0111\u01b0\u1ee3c chi\u1ebfu s\u00e1ng b\u1eb1ng ph\u1ea3n x\u1ea1 to\u00e0n ph\u1ea7n b\u00ean trong (<b>TIRFM<\/b>), qu\u00e9t laze, \u0111a photon v\u00e0 ti\u00eau \u0111i\u1ec3m \u0111\u0129a quay, ngo\u00e0i hu\u1ef3nh quang tr\u01b0\u1eddng r\u1ed9ng truy\u1ec1n th\u1ed1ng.\u00a0C\u00e1c m\u1eabu ph\u00e1t hi\u1ec7n cho t\u1eebng k\u1ef9 thu\u1eadt \u0111\u01b0\u1ee3c bi\u1ec3u th\u1ecb b\u1eb1ng l\u1edbp ph\u1ee7 m\u00e0u \u0111\u1ecf.\u00a0Trong tr\u01b0\u1eddng r\u1ed9ng, m\u1eabu v\u1eadt \u0111\u01b0\u1ee3c chi\u1ebfu s\u00e1ng kh\u1eafp tr\u01b0\u1eddng c\u0169ng nh\u01b0 b\u00ean tr\u00ean v\u00e0 b\u00ean d\u01b0\u1edbi m\u1eb7t ph\u1eb3ng ti\u00eau \u0111i\u1ec3m.\u00a0M\u1ed7i ngu\u1ed3n \u0111i\u1ec3m \u0111\u01b0\u1ee3c tr\u1ea3i th\u00e0nh m\u1ed9t h\u00ecnh gi\u1ed1ng nh\u01b0 m\u1ed9t h\u00ecnh n\u00f3n \u0111\u1ea3o ng\u01b0\u1ee3c \u0111\u01b0\u1ee3c g\u1ecdi l\u00e0 h\u00e0m tr\u1ea3i \u0111i\u1ec3m (<b>PSF<\/b>).\u00a0Ch\u1ec9 ph\u1ea7n trung t\u00e2m c\u1ee7a h\u00ecnh d\u1ea1ng n\u00e0y n\u1eb1m trong m\u1eb7t ph\u1eb3ng ti\u00eau c\u1ef1 v\u1edbi ph\u1ea7n c\u00f2n l\u1ea1i g\u00f3p ph\u1ea7n t\u1ea1o ra hi\u1ec7n t\u01b0\u1ee3ng m\u1edd ngo\u00e0i ti\u00eau \u0111i\u1ec3m, l\u00e0m gi\u1ea3m ch\u1ea5t l\u01b0\u1ee3ng h\u00ecnh \u1ea3nh.<\/p>\n<p>Ng\u01b0\u1ee3c l\u1ea1i, c\u00e1c k\u00ednh hi\u1ec3n vi \u0111\u1ed3ng ti\u00eau qu\u00e9t laze, \u0111a photon v\u00e0 \u0111\u0129a quay qu\u00e9t m\u1eabu v\u1eadt b\u1eb1ng \u0111\u00e8n ph\u00f3ng \u0111i\u1ec7n h\u1ed3 quang ho\u1eb7c laze h\u1ed9i t\u1ee5 ch\u1eb7t ch\u1ebd (\u0111\u0129a quay). M\u00f4 h\u00ecnh k\u00edch th\u00edch l\u00e0 m\u1ed9t ch\u1ee9c n\u0103ng tr\u1ea3i \u0111i\u1ec3m, nh\u01b0ng m\u1ed9t l\u1ed7 kim li\u00ean h\u1ee3p trong \u0111\u01b0\u1eddng quang c\u1ee7a k\u00ednh hi\u1ec3n vi \u0111\u1ed3ng ti\u00eau ng\u0103n kh\u00f4ng cho hu\u1ef3nh quang ph\u00e1t ra t\u1eeb m\u1eb7t ph\u1eb3ng ti\u00eau \u0111i\u1ec3m t\u00e1c \u0111\u1ed9ng \u0111\u1ebfn b\u1ed9 nh\u00e2n quang ho\u1eb7c m\u00e1y d\u00f2 m\u00e1y \u1ea3nh k\u1ef9 thu\u1eadt s\u1ed1. K\u00ednh hi\u1ec3n vi \u0111\u1ed3ng ti\u00eau qu\u00e9t laze c\u00f3 m\u1ed9t \u0111i\u1ec3m laze h\u1ed9i t\u1ee5 duy nh\u1ea5t \u0111\u01b0\u1ee3c qu\u00e9t qua m\u1eabu v\u1eadt. Trong k\u00ednh hi\u1ec3n vi \u0111\u0129a quay, m\u1ed9t d\u00e3y c\u00e1c khe h\u1edf, trong m\u1ed9t s\u1ed1 tr\u01b0\u1eddng h\u1ee3p \u0111\u01b0\u1ee3c g\u1eafn c\u00e1c ph\u1ea7n t\u1eed th\u1ea5u k\u00ednh si\u00eau nh\u1ecf, \u0111\u01b0\u1ee3c \u0111\u1eb7t tr\u00ean m\u1ed9t \u0111\u0129a quay sao cho c\u00e1c l\u1ed7 n\u00e0y nhanh ch\u00f3ng qu\u00e9t qua m\u1eabu v\u1eadt v\u00e0 t\u1ea1o ra h\u00ecnh \u1ea3nh \u0111\u01b0\u1ee3c ghi l\u1ea1i b\u1eb1ng m\u00e1y \u1ea3nh k\u1ef9 thu\u1eadt s\u1ed1. Trong k\u00ednh hi\u1ec3n vi \u0111a photon, v\u00f9ng m\u00e0 t\u1ea1i \u0111\u00f3 th\u00f4ng l\u01b0\u1ee3ng photon \u0111\u1ee7 cao \u0111\u1ec3 k\u00edch th\u00edch c\u00e1c hu\u1ef3nh quang c\u00f3 nhi\u1ec1u h\u01a1n m\u1ed9t photon n\u1eb1m \u1edf v\u1ecb tr\u00ed \u0111\u00fang n\u00e9t c\u1ee7a ch\u1ee9c n\u0103ng tr\u1ea3i \u0111i\u1ec3m. Do \u0111\u00f3, k\u00edch th\u00edch fluorophore ch\u1ec9 x\u1ea3y ra trong ti\u00eau \u0111i\u1ec3m. B\u1edfi v\u00ec t\u1ea5t c\u1ea3 hu\u1ef3nh quang ph\u00e1t ra t\u1eeb c\u00e1c hu\u1ef3nh quang \u0111\u00fang ti\u00eau \u0111i\u1ec3m, kh\u00f4ng c\u1ea7n l\u1ed7 kim v\u00e0 hu\u1ef3nh quang ph\u00e1t ra t\u1ea1o ra h\u00ecnh \u1ea3nh s\u1eafc n\u00e9t, \u0111\u00fang ti\u00eau \u0111i\u1ec3m.<\/p>\n<h2 class=\"paratitle\"><span class=\"ez-toc-section\" id=\"Kinh_hien_vi_huynh_quang_va_ky_thuat_truyen_sang\"><\/span><strong><a href=\"https:\/\/vospan.com\/product-brands\/zeiss-vi\/\">K\u00ednh hi\u1ec3n vi hu\u1ef3nh quang<\/a> v\u00e0 k\u1ef9 thu\u1eadt truy\u1ec1n s\u00e1ng<\/strong><span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p>M\u1ed9t trong nh\u1eefng k\u1ef9 thu\u1eadt ch\u00ednh \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng trong t\u1ea5t c\u1ea3 c\u00e1c d\u1ea1ng k\u00ednh hi\u1ec3n vi hu\u1ef3nh quang l\u00e0\u00a0<b>tr\u01b0\u1eddng s\u00e1ng <\/b>chi\u1ebfu s\u00e1ng d\u1ef1a tr\u00ean nh\u1eefng thay \u0111\u1ed5i v\u1ec1 \u0111\u1ed9 h\u1ea5p th\u1ee5 \u00e1nh s\u00e1ng, ch\u1ec9 s\u1ed1 kh\u00fac x\u1ea1 ho\u1eb7c m\u00e0u s\u1eafc \u0111\u1ec3 t\u1ea1o ra \u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n. Khi \u00e1nh s\u00e1ng \u0111i qua m\u1eabu v\u1eadt, c\u00e1c v\u00f9ng l\u00e0m thay \u0111\u1ed5i h\u01b0\u1edbng, t\u1ed1c \u0111\u1ed9 v\u00e0\/ho\u1eb7c quang ph\u1ed5 c\u1ee7a m\u1eb7t s\u00f3ng t\u1ea1o ra s\u1ef1 ch\u00eanh l\u1ec7ch hu\u1ef3nh quang (\u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n) khi c\u00e1c tia \u0111\u01b0\u1ee3c v\u1eadt k\u00ednh h\u1ed9i t\u1ee5 v\u00e0 h\u1ed9i t\u1ee5. \u0110\u1ed9 ph\u00e2n gi\u1ea3i trong h\u1ec7 th\u1ed1ng tr\u01b0\u1eddng s\u00e1ng ph\u1ee5 thu\u1ed9c v\u00e0o c\u1ea3 kh\u1ea9u \u0111\u1ed9 s\u1ed1 c\u1ee7a v\u1eadt k\u00ednh v\u00e0 t\u1ee5 quang, v\u00e0 m\u00f4i tr\u01b0\u1eddng ng\u00e2m th\u01b0\u1eddng \u0111\u01b0\u1ee3c y\u00eau c\u1ea7u \u1edf c\u1ea3 hai m\u1eb7t c\u1ee7a m\u1eabu v\u1eadt (\u0111\u1ed1i v\u1edbi c\u00e1c t\u1ed5 h\u1ee3p kh\u1ea9u \u0111\u1ed9 s\u1ed1 v\u01b0\u1ee3t qu\u00e1 gi\u00e1 tr\u1ecb 1,0). M\u00e1y \u1ea3nh k\u1ef9 thu\u1eadt s\u1ed1 cung c\u1ea5p d\u1ea3i \u0111\u1ed9ng r\u1ed9ng v\u00e0 \u0111\u1ed9 ph\u00e2n gi\u1ea3i kh\u00f4ng gian c\u1ea7n thi\u1ebft \u0111\u1ec3 n\u1eafm b\u1eaft th\u00f4ng tin c\u00f3 trong h\u00ecnh \u1ea3nh tr\u01b0\u1eddng s\u00e1ng. Ngo\u00e0i ra, c\u00e1c thu\u1eadt to\u00e1n tr\u1eeb n\u1ec1n, s\u1eed d\u1ee5ng c\u00e1c khung l\u1ea5y trung b\u00ecnh \u0111\u01b0\u1ee3c ch\u1ee5p m\u00e0 kh\u00f4ng c\u00f3 m\u1eabu v\u1eadt trong \u0111\u01b0\u1eddng d\u1eabn hu\u1ef3nh quang l\u00e0m t\u0103ng \u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n \u0111\u00e1ng k\u1ec3.<\/p>\n<p><img decoding=\"async\" class=\"aligncenter\" src=\"https:\/\/zeiss-campus.magnet.fsu.edu\/articles\/livecellimaging\/images\/techniquesfigure2.jpg\" \/><\/p>\n<p>H\u00ecnh \u1ea3nh tr\u01b0\u1eddng s\u00e1ng \u0111\u01a1n gi\u1ea3n, v\u1edbi k\u00ednh hi\u1ec3n vi \u0111\u01b0\u1ee3c \u0111i\u1ec1u ch\u1ec9nh ph\u00f9 h\u1ee3p \u0111\u1ec3 chi\u1ebfu s\u00e1ng K\u00f6hler (xem <em>H\u00ecnh 2(a)<\/em>), cung c\u1ea5p m\u1ed9t l\u01b0\u1ee3ng th\u00f4ng tin h\u1ea1n ch\u1ebf v\u1ec1 \u0111\u01b0\u1eddng vi\u1ec1n t\u1ebf b\u00e0o, v\u1ecb tr\u00ed h\u1ea1t nh\u00e2n v\u00e0 v\u1ecb tr\u00ed c\u1ee7a c\u00e1c t\u00fai l\u1edbn h\u01a1n. Tuy nhi\u00ean, vi\u1ec7c thi\u1ebfu \u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n chung trong ch\u1ebf \u0111\u1ed9 tr\u01b0\u1eddng s\u00e1ng khi\u1ebfn k\u1ef9 thu\u1eadt n\u00e0y t\u01b0\u01a1ng \u0111\u1ed1i v\u00f4 d\u1ee5ng \u0111\u1ed1i v\u1edbi c\u00e1c nghi\u00ean c\u1ee9u nghi\u00eam t\u00fac v\u1ec1 c\u1ea5u tr\u00fac v\u00e0 ch\u1ee9c n\u0103ng c\u1ee7a t\u1ebf b\u00e0o. C\u00e1c ph\u01b0\u01a1ng ph\u00e1p t\u0103ng c\u01b0\u1eddng \u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n bao g\u1ed3m \u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n nhi\u1ec5u vi sai (<b>DIC<\/b>), \u00e1nh s\u00e1ng ph\u00e2n c\u1ef1c, \u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n pha, \u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n \u0111i\u1ec1u ch\u1ebf Hoffman v\u00e0 k\u00ednh hi\u1ec3n vi tr\u01b0\u1eddng t\u1ed1i (c\u00e1c v\u00ed d\u1ee5 \u0111\u01b0\u1ee3c minh h\u1ecda trong <em>H\u00ecnh 2<\/em>). M\u1ed9t s\u1ed1 k\u1ef9 thu\u1eadt trong s\u1ed1 n\u00e0y b\u1ecb gi\u1edbi h\u1ea1n b\u1edfi \u00e1nh s\u00e1ng b\u1eaft ngu\u1ed3n t\u1eeb c\u00e1c v\u00f9ng b\u1ecb lo\u1ea1i b\u1ecf kh\u1ecfi m\u1eb7t ph\u1eb3ng ti\u00eau \u0111i\u1ec3m khi ch\u1ee5p \u1ea3nh c\u00e1c m\u00f4 \u0111\u1ed9ng v\u1eadt v\u00e0 th\u1ef1c v\u1eadt d\u00e0y h\u01a1n, trong khi \u00e1nh s\u00e1ng ph\u00e2n c\u1ef1c y\u00eau c\u1ea7u t\u00ednh l\u01b0\u1ee1ng chi\u1ebft (th\u01b0\u1eddng kh\u00f4ng xu\u1ea5t hi\u1ec7n \u1edf m\u1ee9c \u0111\u1ed9 \u0111\u00e1ng k\u1ec3 trong t\u1ebf b\u00e0o \u0111\u1ed9ng v\u1eadt) \u0111\u1ec3 t\u1ea1o ra \u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n. C\u00e1c t\u1ebf b\u00e0o bu\u1ed3ng tr\u1ee9ng c\u1ee7a chu\u1ed9t \u0111\u1ed3ng Trung Qu\u1ed1c k\u1ebft d\u00ednh \u0111\u01b0\u1ee3c tr\u00ecnh b\u00e0y trong <em>H\u00ecnh 2(a)<\/em> \u0111\u01b0\u1ee3c t\u1ea1o \u1ea3nh trong \u0111i\u1ec1u ki\u1ec7n chi\u1ebfu s\u00e1ng tr\u01b0\u1eddng s\u00e1ng m\u00e0 kh\u00f4ng c\u00f3 s\u1ef1 h\u1ed7 tr\u1ee3 c\u1ee7a ph\u01b0\u01a1ng ph\u00e1p t\u0103ng c\u01b0\u1eddng \u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n hu\u1ef3nh quang. L\u01b0u \u00fd s\u1ef1 thi\u1ebfu t\u01b0\u01a1ng ph\u1ea3n chung khi so s\u00e1nh v\u1edbi c\u00e1c h\u00ecnh \u1ea3nh kh\u00e1c trong h\u00ecnh. C\u00e1c t\u1ebf b\u00e0o ri\u00eang l\u1ebb r\u1ea5t kh\u00f3 ph\u00e2n bi\u1ec7t v\u00e0 h\u1ea7u h\u1ebft c\u00e1c t\u00ednh n\u0103ng b\u00ean trong (v\u00ed d\u1ee5: h\u1ea1t nh\u00e2n) kh\u00f4ng hi\u1ec3n th\u1ecb trong h\u00ecnh \u1ea3nh n\u00e0y.<\/p>\n<p><b>S\u1ef1 t\u01b0\u01a1ng ph\u1ea3n ch\u00eanh l\u1ec7ch <\/b>c\u1ee7a\u00a0k\u00ednh hi\u1ec3n vi (<em>H\u00ecnh 2(b)<\/em> v\u00e0 <em>H\u00ecnh 3(a)<\/em>) y\u00eau c\u1ea7u \u00e1nh s\u00e1ng ph\u00e2n c\u1ef1c ph\u1eb3ng v\u00e0 c\u00e1c l\u0103ng k\u00ednh c\u1eaft \u00e1nh s\u00e1ng b\u1ed5 sung (Nomarski ho\u1eb7c Wollaston) \u0111\u1ec3 ph\u00f3ng \u0111\u1ea1i s\u1ef1 kh\u00e1c bi\u1ec7t nh\u1ecf v\u1ec1 \u0111\u1ed9 d\u1ed1c \u0111\u1ed9 d\u00e0y m\u1eabu v\u1eadt v\u00e0 ch\u1ec9 s\u1ed1 kh\u00fac x\u1ea1.\u00a0V\u00ed d\u1ee5, hai l\u1edbp lipid t\u1ea1o ra \u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n tuy\u1ec7t v\u1eddi trong DIC do s\u1ef1 kh\u00e1c bi\u1ec7t v\u1ec1 ch\u1ec9 s\u1ed1 kh\u00fac x\u1ea1 gi\u1eefa c\u00e1c pha n\u01b0\u1edbc v\u00e0 lipid c\u1ee7a t\u1ebf b\u00e0o.\u00a0Ngo\u00e0i ra, ranh gi\u1edbi t\u1ebf b\u00e0o trong c\u00e1c t\u1ebf b\u00e0o th\u1ef1c v\u1eadt v\u00e0 \u0111\u1ed9ng v\u1eadt c\u00f3 v\u00fa t\u01b0\u01a1ng \u0111\u1ed1i ph\u1eb3ng, bao g\u1ed3m m\u00e0ng sinh ch\u1ea5t, nh\u00e2n, kh\u00f4ng b\u00e0o, ty th\u1ec3 v\u00e0 c\u00e1c s\u1ee3i \u1ee9ng su\u1ea5t, th\u01b0\u1eddng t\u1ea1o ra \u0111\u1ed9 d\u1ed1c \u0111\u00e1ng k\u1ec3, d\u1ec5 d\u00e0ng \u0111\u01b0\u1ee3c t\u1ea1o \u1ea3nh b\u1eb1ng DIC.\u00a0Trong c\u00e1c m\u00f4 th\u1ef1c v\u1eadt, th\u00e0nh t\u1ebf b\u00e0o l\u01b0\u1ee1ng chi\u1ebft l\u00e0m gi\u1ea3m \u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n trong DIC \u1edf m\u1ed9t m\u1ee9c \u0111\u1ed9 h\u1ea1n ch\u1ebf, nh\u01b0ng m\u1ed9t h\u1ec7 th\u1ed1ng \u0111\u01b0\u1ee3c c\u0103n ch\u1ec9nh ph\u00f9 h\u1ee3p s\u1ebd cho ph\u00e9p h\u00ecnh dung m\u00e0ng nh\u00e2n v\u00e0 m\u00e0ng kh\u00f4ng b\u00e0o, ty th\u1ec3, l\u1ee5c l\u1ea1p,\u00a0v\u00e0 nhi\u1ec5m s\u1eafc th\u1ec3 ng\u01b0ng t\u1ee5 trong c\u00e1c t\u1ebf b\u00e0o bi\u1ec3u b\u00ec.\u00a0\u0110\u1ed9 t\u01b0\u01a1ng ph\u1ea3n giao thoa vi sai l\u00e0 m\u1ed9t k\u1ef9 thu\u1eadt quan tr\u1ecdng \u0111\u1ec3 ch\u1ee5p \u1ea3nh c\u00e1c m\u00f4 \u0111\u1ed9ng v\u1eadt v\u00e0 th\u1ef1c v\u1eadt d\u00e0y v\u00ec ngo\u00e0i \u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n t\u0103ng l\u00ean, DIC c\u00f2n th\u1ec3 hi\u1ec7n \u0111\u1ed9 s\u00e2u ti\u00eau c\u1ef1 gi\u1ea3m \u1edf kh\u1ea9u \u0111\u1ed9 r\u1ed9ng, t\u1ea1o ra ti\u1ebft di\u1ec7n hu\u1ef3nh quang m\u1ecfng c\u1ee7a m\u1eabu v\u1eadt d\u00e0y.\u00a0Hi\u1ec7u \u1ee9ng n\u00e0y c\u0169ng thu\u1eadn l\u1ee3i cho vi\u1ec7c ch\u1ee5p \u1ea3nh c\u00e1c t\u1ebf b\u00e0o k\u1ebft d\u00ednh \u0111\u1ec3 gi\u1ea3m thi\u1ec3u hi\u1ec7n t\u01b0\u1ee3ng m\u1edd ph\u00e1t sinh t\u1eeb c\u00e1c m\u1ea3nh v\u1ee5n tr\u00f4i n\u1ed5i trong m\u00f4i tr\u01b0\u1eddng nu\u00f4i c\u1ea5y.<\/p>\n<p><img decoding=\"async\" class=\"aligncenter\" src=\"https:\/\/zeiss-campus.magnet.fsu.edu\/articles\/livecellimaging\/images\/techniquesfigure3.jpg\" \/><\/p>\n<p><b>\u00c1nh s\u00e1ng ph\u00e2n c\u1ef1c <\/b>c\u1ee7a\u00a0k\u00ednh hi\u1ec3n vi (<em>H\u00ecnh 2(f)<\/em> v\u00e0 <em>H\u00ecnh 3(b)<\/em>) \u0111\u01b0\u1ee3c ti\u1ebfn h\u00e0nh b\u1eb1ng c\u00e1ch xem m\u1eabu gi\u1eefa c\u00e1c ph\u1ea7n t\u1eed ph\u00e2n c\u1ef1c ch\u00e9o.\u00a0C\u00e1c t\u1eadp h\u1ee3p b\u00ean trong t\u1ebf b\u00e0o c\u00f3 \u0111\u1eb7c t\u00ednh l\u01b0\u1ee1ng chi\u1ebft, ch\u1eb3ng h\u1ea1n nh\u01b0 th\u00e0nh t\u1ebf b\u00e0o th\u1ef1c v\u1eadt, h\u1ea1t tinh b\u1ed9t v\u00e0 thoi ph\u00e2n b\u00e0o, c\u0169ng nh\u01b0 m\u00f4 c\u01a1, l\u00e0m quay m\u1eb7t ph\u1eb3ng ph\u00e2n c\u1ef1c \u00e1nh s\u00e1ng, xu\u1ea5t hi\u1ec7n s\u00e1ng tr\u00ean n\u1ec1n t\u1ed1i.\u00a0M\u00f4 c\u01a1 th\u1ecf \u0111\u01b0\u1ee3c minh h\u1ecda trong H\u00ecnh 2(f) l\u00e0 m\u1ed9t v\u00ed d\u1ee5 v\u1ec1 k\u00ednh hi\u1ec3n vi \u00e1nh s\u00e1ng ph\u00e2n c\u1ef1c \u0111\u01b0\u1ee3c \u00e1p d\u1ee5ng \u0111\u1ec3 quan s\u00e1t m\u00f4 s\u1ed1ng.\u00a0L\u01b0u \u00fd r\u1eb1ng k\u1ef9 thu\u1eadt n\u00e0y b\u1ecb h\u1ea1n ch\u1ebf b\u1edfi s\u1ef1 xu\u1ea5t hi\u1ec7n l\u01b0\u1ee1ng chi\u1ebft su\u1ea5t hi\u1ebfm g\u1eb7p trong c\u00e1c t\u1ebf b\u00e0o v\u00e0 m\u00f4 s\u1ed1ng v\u00e0 v\u1eabn ch\u01b0a \u0111\u01b0\u1ee3c kh\u00e1m ph\u00e1 \u0111\u1ea7y \u0111\u1ee7.\u00a0Nh\u01b0 \u0111\u00e3 \u0111\u1ec1 c\u1eadp \u1edf tr\u00ean, \u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n giao thoa vi sai ho\u1ea1t \u0111\u1ed9ng b\u1eb1ng c\u00e1ch \u0111\u1eb7t m\u1ed9t c\u1eb7p l\u0103ng k\u00ednh Nomarski \u0111\u1ed1i nhau ph\u00f9 h\u1ee3p gi\u1eefa c\u00e1c b\u1ea3n ph\u00e2n c\u1ef1c ch\u00e9o,<\/p>\n<p><b>\u0110\u1ed9 t\u01b0\u01a1ng ph\u1ea3n pha<\/b> ph\u1ed5 bi\u1ebfn r\u1ed9ng r\u00e3i (nh\u01b0 \u0111\u01b0\u1ee3c minh h\u1ecda trong <em>H\u00ecnh 2(c)<\/em> v\u00e0 <em>H\u00ecnh 4(a)<\/em>) s\u1eed d\u1ee5ng m\u1ed9t c\u01a1 ch\u1ebf hu\u1ef3nh quang \u0111\u1ec3 d\u1ecbch c\u00e1c bi\u1ebfn thi\u00ean nh\u1ecf v\u1ec1 pha th\u00e0nh c\u00e1c thay \u0111\u1ed5i bi\u00ean \u0111\u1ed9 t\u01b0\u01a1ng \u1ee9ng, c\u00f3 th\u1ec3 \u0111\u01b0\u1ee3c h\u00ecnh dung nh\u01b0 l\u00e0 s\u1ef1 kh\u00e1c bi\u1ec7t v\u1ec1 \u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n c\u1ee7a h\u00ecnh \u1ea3nh. K\u00ednh hi\u1ec3n vi ph\u1ea3i \u0111\u01b0\u1ee3c trang b\u1ecb m\u1ed9t t\u1ee5 quang chuy\u00ean d\u1ee5ng ch\u1ee9a m\u1ed9t lo\u1ea1t c\u00e1c v\u00f2ng khuy\u00ean ph\u00f9 h\u1ee3p v\u1edbi m\u1ed9t b\u1ed9 v\u1eadt k\u00ednh ch\u1ee9a c\u00e1c v\u00f2ng pha \u1edf m\u1eb7t ph\u1eb3ng ti\u00eau \u0111i\u1ec3m ph\u00eda sau (c\u00e1c v\u1eadt k\u00ednh t\u01b0\u01a1ng ph\u1ea3n pha c\u0169ng c\u00f3 th\u1ec3 \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng v\u1edbi hu\u1ef3nh quang, nh\u01b0ng \u0111\u1ed9 truy\u1ec1n qua gi\u1ea3m \u0111i m\u1ed9t ch\u00fat). T\u01b0\u01a1ng ph\u1ea3n pha l\u00e0 m\u1ed9t ph\u01b0\u01a1ng ph\u00e1p tuy\u1ec7t v\u1eddi \u0111\u1ec3 t\u0103ng \u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n khi xem ho\u1eb7c ch\u1ee5p \u1ea3nh c\u00e1c t\u1ebf b\u00e0o s\u1ed1ng trong qu\u00e1 tr\u00ecnh nu\u00f4i c\u1ea5y, nh\u01b0ng th\u01b0\u1eddng d\u1eabn \u0111\u1ebfn c\u00e1c qu\u1ea7ng s\u00e1ng qu\u00e1 m\u1ee9c xung quanh \u0111\u01b0\u1eddng vi\u1ec1n c\u1ee7a c\u00e1c \u0111\u1eb7c \u0111i\u1ec3m c\u1ea1nh. Nh\u1eefng v\u1ea7ng h\u00e0o quang n\u00e0y l\u00e0 t\u1ea1o t\u00e1c hu\u1ef3nh quang th\u01b0\u1eddng l\u00e0m gi\u1ea3m kh\u1ea3 n\u0103ng hi\u1ec3n th\u1ecb c\u1ee7a c\u00e1c chi ti\u1ebft ranh gi\u1edbi. K\u1ef9 thu\u1eadt n\u00e0y kh\u00f4ng h\u1eefu \u00edch \u0111\u1ed1i v\u1edbi c\u00e1c m\u1eabu v\u1eadt d\u00e0y (ch\u1eb3ng h\u1ea1n nh\u01b0 c\u00e1c ph\u1ea7n m\u00f4 th\u1ef1c v\u1eadt v\u00e0 \u0111\u1ed9ng v\u1eadt) v\u00ec s\u1ef1 l\u1ec7ch pha x\u1ea3y ra \u1edf c\u00e1c v\u00f9ng b\u1ecb lo\u1ea1i kh\u1ecfi m\u1eb7t ph\u1eb3ng ti\u00eau \u0111i\u1ec3m l\u00e0m bi\u1ebfn d\u1ea1ng chi ti\u1ebft h\u00ecnh \u1ea3nh. H\u01a1n n\u1eefa, c\u00e1c m\u1ea3nh v\u1ee5n tr\u00f4i n\u1ed5i v\u00e0 c\u00e1c v\u1eadt th\u1ec3 n\u1eb1m ngo\u00e0i ti\u00eau \u0111i\u1ec3m kh\u00e1c c\u1ea3n tr\u1edf vi\u1ec7c t\u1ea1o \u1ea3nh c\u00e1c t\u1ebf b\u00e0o b\u00e1m d\u00ednh tr\u00ean n\u1eafp tr\u01b0\u1ee3t.<\/p>\n<p>Th\u01b0\u1eddng \u0111\u01b0\u1ee3c g\u1ecdi m\u1ed9t c\u00e1ch \u1ea9n d\u1ee5 l\u00e0 &#8220;poor man&#8217;s DIC&#8221;, <b>\u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n \u0111i\u1ec1u ch\u1ebf Hoffman<\/b> l\u00e0 m\u1ed9t k\u1ef9 thu\u1eadt chi\u1ebfu s\u00e1ng xi\u00ean gi\u00fap t\u0103ng c\u01b0\u1eddng \u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n trong c\u00e1c t\u1ebf b\u00e0o v\u00e0 m\u00f4 s\u1ed1ng b\u1eb1ng c\u00e1ch ph\u00e1t hi\u1ec7n c\u00e1c gradient hu\u1ef3nh quang (xem <em>H\u00ecnh 2(d)<\/em> v\u00e0 <em>H\u00ecnh 4(b)<\/em>).\u00a0C\u1ea5u h\u00ecnh c\u01a1 b\u1ea3n c\u1ee7a k\u00ednh hi\u1ec3n vi bao g\u1ed3m m\u1ed9t b\u1ed9 l\u1ecdc kh\u00f4ng gian bi\u00ean \u0111\u1ed9 hu\u1ef3nh quang, \u0111\u01b0\u1ee3c g\u1ecdi l\u00e0\u00a0<b>b\u1ed9 \u0111i\u1ec1u bi\u1ebfn<\/b>, \u0111\u01b0\u1ee3c l\u1eafp v\u00e0o m\u1eb7t ph\u1eb3ng ti\u00eau c\u1ef1 ph\u00eda sau c\u1ee7a v\u1eadt k\u00ednh. C\u01b0\u1eddng \u0111\u1ed9 \u00e1nh s\u00e1ng \u0111i qua b\u1ed9 bi\u1ebfn \u0111i\u1ec7u thay \u0111\u1ed5i tr\u00ean v\u00e0 d\u01b0\u1edbi gi\u00e1 tr\u1ecb trung b\u00ecnh, theo \u0111\u1ecbnh ngh\u0129a, khi \u0111\u00f3 \u0111\u01b0\u1ee3c cho l\u00e0 \u0111\u00e3 \u0111i\u1ec1u ch\u1ebf. K\u1ebft h\u1ee3p v\u1edbi b\u1ed9 \u0111i\u1ec1u bi\u1ebfn v\u1eadt k\u00ednh l\u00e0 m\u1ed9t khe h\u1edf ngo\u00e0i tr\u1ee5c \u0111\u01b0\u1ee3c \u0111\u1eb7t trong m\u1eb7t ph\u1eb3ng ti\u00eau c\u1ef1 ph\u00eda tr\u01b0\u1edbc c\u1ee7a t\u1ee5 quang \u0111\u1ec3 h\u01b0\u1edbng \u00e1nh s\u00e1ng xi\u00ean v\u1ec1 ph\u00eda m\u1eabu v\u1eadt. Kh\u00f4ng gi\u1ed1ng nh\u01b0 t\u1ea5m pha trong k\u00ednh hi\u1ec3n vi t\u01b0\u01a1ng ph\u1ea3n pha, b\u1ed9 bi\u1ebfn \u0111i\u1ec7u Hoffman \u0111\u01b0\u1ee3c thi\u1ebft k\u1ebf \u0111\u1ec3 kh\u00f4ng l\u00e0m thay \u0111\u1ed5i pha c\u1ee7a \u00e1nh s\u00e1ng \u0111i qua; thay v\u00e0o \u0111\u00f3, n\u00f3 \u1ea3nh h\u01b0\u1edfng \u0111\u1ebfn c\u1ef1c \u0111\u1ea1i b\u1eadc 0 ch\u00ednh \u0111\u1ec3 t\u1ea1o ra \u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n. \u0110\u1ed9 t\u01b0\u01a1ng ph\u1ea3n \u0111i\u1ec1u ch\u1ebf Hoffman kh\u00f4ng b\u1ecb c\u1ea3n tr\u1edf b\u1edfi vi\u1ec7c s\u1eed d\u1ee5ng c\u00e1c v\u1eadt li\u1ec7u l\u01b0\u1ee1ng chi\u1ebft (ch\u1eb3ng h\u1ea1n nh\u01b0 \u0111\u0129a Petri b\u1eb1ng nh\u1ef1a) trong \u0111\u01b0\u1eddng d\u1eabn hu\u1ef3nh quang, v\u00ec v\u1eady k\u1ef9 thu\u1eadt n\u00e0y h\u1eefu \u00edch h\u01a1n \u0111\u1ec3 ki\u1ec3m tra m\u1eabu v\u1eadt trong c\u00e1c v\u1eadt ch\u1ee9a \u0111\u01b0\u1ee3c ch\u1ebf t\u1ea1o b\u1eb1ng v\u1eadt li\u1ec7u polyme. V\u1ec1 m\u1eb7t nh\u01b0\u1ee3c \u0111i\u1ec3m,<\/p>\n<p><img loading=\"lazy\" decoding=\"async\" class=\"aligncenter\" src=\"https:\/\/zeiss-campus.magnet.fsu.edu\/articles\/livecellimaging\/images\/techniquesfigure4.jpg\" width=\"452\" height=\"264\" border=\"0\" \/>Ph\u01b0\u01a1ng ph\u00e1p tr\u01b0\u1eddng t\u1ed1i c\u1ee7a k\u00ednh hi\u1ec3n vi, m\u1eb7c d\u00f9 \u0111\u01b0\u1ee3c \u00e1p d\u1ee5ng r\u1ed9ng r\u00e3i \u0111\u1ec3 ch\u1ee5p \u1ea3nh c\u00e1c m\u1eabu v\u1eadt trong su\u1ed1t trong su\u1ed1t th\u1ebf k\u1ef7 19 v\u00e0 20, nh\u01b0ng b\u1ecb h\u1ea1n ch\u1ebf s\u1eed d\u1ee5ng cho c\u00e1c t\u1ebf b\u00e0o v\u00e0 sinh v\u1eadt b\u1ecb c\u00f4 l\u1eadp v\u1ec1 m\u1eb7t v\u1eadt l\u00fd (nh\u01b0 \u0111\u01b0\u1ee3c tr\u00ecnh b\u00e0y trong <em>H\u00ecnh 2(e)<\/em>). Trong k\u1ef9 thu\u1eadt n\u00e0y, t\u1ee5 quang h\u01b0\u1edbng m\u1ed9t h\u00ecnh n\u00f3n \u00e1nh s\u00e1ng v\u00e0o m\u1eabu v\u1eadt \u1edf c\u00e1c g\u00f3c ph\u01b0\u01a1ng v\u1ecb cao sao cho c\u00e1c m\u1eb7t s\u00f3ng b\u1eadc m\u1ed9t kh\u00f4ng \u0111i tr\u1ef1c ti\u1ebfp v\u00e0o th\u00e0nh ph\u1ea7n th\u1ea5u k\u00ednh ph\u00eda tr\u01b0\u1edbc v\u1eadt k\u00ednh. \u00c1nh s\u00e1ng \u0111i qua m\u1eabu v\u1eadt b\u1ecb nhi\u1ec5u x\u1ea1, ph\u1ea3n x\u1ea1 v\u00e0\/ho\u1eb7c kh\u00fac x\u1ea1 b\u1edfi s\u1ef1 gi\u00e1n \u0111o\u1ea1n quang h\u1ecdc (ch\u1eb3ng h\u1ea1n nh\u01b0 m\u00e0ng t\u1ebf b\u00e0o, nh\u00e2n v\u00e0 c\u00e1c b\u00e0o quan b\u00ean trong) cho ph\u00e9p c\u00e1c tia y\u1ebfu n\u00e0y \u0111i v\u00e0o v\u1eadt k\u00ednh. M\u1eabu v\u1eadt sau \u0111\u00f3 c\u00f3 th\u1ec3 \u0111\u01b0\u1ee3c h\u00ecnh dung nh\u01b0 m\u1ed9t v\u1eadt th\u1ec3 s\u00e1ng tr\u00ean n\u1ec1n \u0111en. Th\u1eadt kh\u00f4ng may, \u00e1nh s\u00e1ng t\u00e1n x\u1ea1 b\u1edfi c\u00e1c v\u1eadt th\u1ec3 b\u1ecb lo\u1ea1i b\u1ecf kh\u1ecfi m\u1eb7t ph\u1eb3ng ti\u00eau c\u1ef1 c\u0169ng g\u00f3p ph\u1ea7n t\u1ea1o n\u00ean h\u00ecnh \u1ea3nh, do \u0111\u00f3 l\u00e0m gi\u1ea3m \u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n v\u00e0 che khu\u1ea5t chi ti\u1ebft m\u1eabu v\u1eadt. Hi\u1ec7n t\u01b0\u1ee3ng n\u00e0y \u0111\u01b0\u1ee3c k\u1ebft h\u1ee3p b\u1edfi th\u1ef1c t\u1ebf l\u00e0 b\u1ee5i v\u00e0 m\u1ea3nh v\u1ee5n trong bu\u1ed3ng t\u1ea1o \u1ea3nh c\u0169ng g\u00f3p ph\u1ea7n \u0111\u00e1ng k\u1ec3 v\u00e0o h\u00ecnh \u1ea3nh thu \u0111\u01b0\u1ee3c. H\u01a1n n\u1eefa, c\u00e1c t\u1ebf b\u00e0o k\u1ebft d\u00ednh m\u1ecfng th\u01b0\u1eddng ch\u1ecbu t\u00edn hi\u1ec7u r\u1ea5t y\u1ebfu, trong khi c\u00e1c m\u00f4 th\u1ef1c v\u1eadt v\u00e0 \u0111\u1ed9ng v\u1eadt d\u00e0y chuy\u1ec3n h\u01b0\u1edbng qu\u00e1 nhi\u1ec1u \u00e1nh s\u00e1ng v\u00e0o \u0111\u01b0\u1eddng d\u1eabn m\u1ee5c ti\u00eau, l\u00e0m gi\u1ea3m hi\u1ec7u qu\u1ea3 c\u1ee7a k\u1ef9 thu\u1eadt.<\/p>\n<h2><span class=\"ez-toc-section\" id=\"Kinh_hien_vi_huynh_quang_truong_rong\"><\/span><strong><a href=\"https:\/\/vospan.com\/product-brands\/zeiss-vi\/\">K\u00ednh hi\u1ec3n vi hu\u1ef3nh quang<\/a> tr\u01b0\u1eddng r\u1ed9ng<\/strong><span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p>Trong c\u00e1c \u1ee9ng d\u1ee5ng ch\u1ee5p \u1ea3nh t\u1ebf b\u00e0o s\u1ed1ng, k\u00ednh hi\u1ec3n\u00a0<b>vi hu\u1ef3nh quang tr\u01b0\u1eddng r\u1ed9ng<\/b> r\u1ea5t h\u1eefu \u00edch \u0111\u1ec3 quan s\u00e1t \u0111\u1ed9ng l\u1ef1c h\u1ecdc c\u1ee7a c\u00e1c t\u1ebf b\u00e0o k\u1ebft d\u00ednh ph\u00e1t tri\u1ec3n trong c\u00e1c bu\u1ed3ng m\u00f4i tr\u01b0\u1eddng chuy\u00ean bi\u1ec7t th\u00edch \u1ee9ng v\u1edbi giai \u0111o\u1ea1n k\u00ednh hi\u1ec3n vi. \u1ede c\u1ea5u h\u00ecnh th\u00f4 s\u01a1 nh\u1ea5t, k\u00ednh hi\u1ec3n vi nu\u00f4i c\u1ea5y m\u00f4 \u0111\u1ea3o ng\u01b0\u1ee3c ti\u00eau chu\u1ea9n \u0111\u01b0\u1ee3c trang b\u1ecb chi\u1ebfu s\u00e1ng hu\u1ef3nh quang epi \u0111\u01b0\u1ee3c k\u1ebft h\u1ee3p v\u1edbi h\u1ec7 th\u1ed1ng m\u00e1y \u1ea3nh CCD, b\u1ed9 l\u1ecdc hu\u1ef3nh quang th\u00edch h\u1ee3p v\u00e0 h\u1ec7 th\u1ed1ng m\u00e0n tr\u1eadp \u0111\u1ec3 h\u1ea1n ch\u1ebf s\u1ef1 ph\u01a1i nhi\u1ec5m qu\u00e1 m\u1ee9c c\u1ee7a t\u1ebf b\u00e0o v\u1edbi \u00e1nh s\u00e1ng k\u00edch th\u00edch c\u00f3 h\u1ea1i. K\u00ednh hi\u1ec3n vi hu\u1ef3nh quang c\u01a1 b\u1ea3n d\u1ef1a v\u00e0o c\u00e1c b\u1ed9 l\u1ecdc nhi\u1ec5u ph\u00f9 h\u1ee3p c\u1ea9n th\u1eadn trong vi\u1ec7c l\u1ef1a ch\u1ecdn b\u0103ng th\u00f4ng c\u1ee5 th\u1ec3 \u0111\u1ec3 chi\u1ebfu s\u00e1ng v\u00e0 ph\u00e1t hi\u1ec7n ph\u00e1t x\u1ea1. C\u00e1c ngu\u1ed3n s\u00e1ng bao g\u1ed3m \u0111\u00e8n h\u1ed3 quang th\u1ee7y ng\u00e2n, xenon v\u00e0 halogen kim lo\u1ea1i, h\u1ec7 th\u1ed1ng laze m\u1edf r\u1ed9ng ch\u00f9m tia v\u00e0 \u0111i\u1ed1t ph\u00e1t quang (<b>LED<\/b>), t\u1ea5t c\u1ea3 \u0111\u1ec1u y\u00eau c\u1ea7u c\u00e1c th\u00f4ng s\u1ed1 k\u1ef9 thu\u1eadt b\u1ed9 l\u1ecdc kh\u00e1c nhau.\u00a0C\u00e1c hu\u1ef3nh quang t\u1ed5ng h\u1ee3p h\u1eefu \u00edch trong k\u00ednh hi\u1ec3n vi hu\u1ef3nh quang c\u00f3 ph\u1ed5 ph\u00e1t x\u1ea1 bao ph\u1ee7 c\u00e1c v\u00f9ng c\u1eadn t\u1eed ngo\u1ea1i, kh\u1ea3 ki\u1ebfn \u200b\u200bv\u00e0 c\u1eadn h\u1ed3ng ngo\u1ea1i.\u00a0Vi\u1ec7c \u00e1p d\u1ee5ng c\u00e1c protein hu\u1ef3nh quang \u0111\u01b0\u1ee3c m\u00e3 h\u00f3a di truy\u1ec1n \u0111\u00e3 m\u1edf r\u1ed9ng \u0111\u00e1ng k\u1ec3 kh\u1ea3 n\u0103ng ch\u1ee5p \u1ea3nh t\u1ebf b\u00e0o s\u1ed1ng trong k\u00ednh hi\u1ec3n vi hu\u1ef3nh quang b\u1eb1ng c\u00e1ch cho ph\u00e9p c\u00e1c nh\u00e0 \u0111i\u1ec1u tra nh\u1eafm m\u1ee5c ti\u00eau v\u00e0o c\u00e1c v\u00f9ng quan t\u00e2m d\u01b0\u1edbi t\u1ebf b\u00e0o v\u1edbi \u0111\u1ed9 ch\u00ednh x\u00e1c ch\u00ednh x\u00e1c.<\/p>\n<p>Trong hu\u1ef3nh quang tr\u01b0\u1eddng r\u1ed9ng, \u0111\u1ed9 m\u1edf \u0111\u1ea7y \u0111\u1ee7 c\u1ee7a \u00e1nh s\u00e1ng ph\u00e1t x\u1ea1 \u0111\u01b0\u1ee3c thu th\u1eadp b\u1edfi v\u1eadt k\u00ednh c\u1ee7a k\u00ednh hi\u1ec3n vi s\u1ebd t\u1ed1i \u0111a h\u00f3a t\u00edn hi\u1ec7u \u0111\u01b0\u1ee3c ghi l\u1ea1i v\u00e0 \u0111\u1ed3ng th\u1eddi gi\u1ea3m thi\u1ec3u th\u1eddi gian ph\u01a1i s\u00e1ng c\u1ea7n thi\u1ebft. Do \u0111\u00f3, c\u00e1c m\u1eabu v\u1eadt c\u00f3 th\u1ec3 \u0111\u01b0\u1ee3c ch\u1ee5p \u1ea3nh v\u1edbi th\u1eddi gian chi\u1ebfu s\u00e1ng r\u1ea5t ng\u1eafn. H\u1ea1n ch\u1ebf ch\u00ednh \u0111\u1ed1i v\u1edbi h\u00ecnh \u1ea3nh tr\u01b0\u1eddng r\u1ed9ng l\u00e0 hu\u1ef3nh quang ph\u00e1t sinh t\u1eeb c\u00e1c v\u00f9ng c\u00e1ch xa m\u1eb7t ph\u1eb3ng ti\u00eau c\u1ef1, c\u0169ng nh\u01b0 t\u00edn hi\u1ec7u n\u1ec1n, t\u1ea1o ra \u00e1nh s\u00e1ng kh\u00f4ng h\u1eefu \u00edch th\u01b0\u1eddng che khu\u1ea5t c\u00e1c \u0111\u1eb7c \u0111i\u1ec3m quan t\u00e2m. C\u00f3 r\u1ea5t nhi\u1ec1u m\u1eabu t\u1ebf b\u00e0o s\u1ed1ng, bao g\u1ed3m c\u00e1c t\u1ebf b\u00e0o k\u1ebft d\u00ednh, vi khu\u1ea9n, n\u1ea5m men v\u00e0 c\u00e1c ph\u1ea7n m\u00f4 r\u1ea5t m\u1ecfng, l\u00e0 nh\u1eefng m\u1eabu l\u00fd t\u01b0\u1edfng \u0111\u1ec3 t\u1ea1o \u1ea3nh trong hu\u1ef3nh quang tr\u01b0\u1eddng r\u1ed9ng.<\/p>\n<p><img decoding=\"async\" class=\"aligncenter\" src=\"https:\/\/zeiss-campus.magnet.fsu.edu\/articles\/livecellimaging\/images\/techniquesfigure5.jpg\" \/><\/p>\n<p>M\u1eb7c d\u00f9 \u0111\u00e3 c\u00f3 nhi\u1ec1u ti\u1ebfn b\u1ed9 trong vi\u1ec7c ph\u00e1t tri\u1ec3n thu\u1ed1c nhu\u1ed9m hu\u1ef3nh quang t\u1ed5ng h\u1ee3p, ch\u1ea5m l\u01b0\u1ee3ng t\u1eed v\u00e0 protein hu\u1ef3nh quang \u0111\u1ec3 ch\u1ee5p \u1ea3nh \u1edf ch\u1ebf \u0111\u1ed9 hu\u1ef3nh quang, nh\u01b0ng trong m\u1ed9t s\u1ed1 tr\u01b0\u1eddng h\u1ee3p, vi\u1ec7c k\u1ebft h\u1ee3p hu\u1ef3nh quang v\u1edbi c\u00e1c ph\u01b0\u01a1ng th\u1ee9c ch\u1ee5p \u1ea3nh kh\u00e1c l\u00e0 thu\u1eadn l\u1ee3i.\u00a0V\u00ed d\u1ee5, k\u00ednh hi\u1ec3n vi DIC c\u00f3 th\u1ec3 \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng c\u00f9ng v\u1edbi hu\u1ef3nh quang tr\u01b0\u1eddng r\u1ed9ng (H\u00ecnh 5(c)) \u0111\u1ec3 theo d\u00f5i kh\u1ea3 n\u0103ng t\u1ed3n t\u1ea1i c\u1ee7a t\u1ebf b\u00e0o v\u00e0 h\u00ecnh th\u00e1i chung \u0111\u1ed3ng th\u1eddi \u0111i\u1ec1u tra c\u00e1c hi\u1ec7n t\u01b0\u1ee3ng quan t\u00e2m v\u1edbi c\u00e1c m\u1ee5c ti\u00eau \u0111\u01b0\u1ee3c d\u00e1n nh\u00e3n c\u1ee5 th\u1ec3.\u00a0Ch\u1ee5p DIC v\u00e0 hu\u1ef3nh quang trong m\u1ed9t khung h\u00ecnh duy nh\u1ea5t th\u01b0\u1eddng kh\u00f4ng th\u1ef1c t\u1ebf, nh\u01b0ng trong k\u00ednh hi\u1ec3n vi \u0111\u01b0\u1ee3c c\u1ea5u h\u00ecnh ph\u00f9 h\u1ee3p, hai k\u1ef9 thu\u1eadt n\u00e0y c\u00f3 th\u1ec3 \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng tu\u1ea7n t\u1ef1.\u00a0Do \u0111\u00f3, h\u00ecnh \u1ea3nh DIC c\u00f3 th\u1ec3 \u0111\u01b0\u1ee3c ch\u1ee5p t\u1eeb m\u1ed9t m\u1eabu \u0111\u01b0\u1ee3c d\u00e1n nh\u00e3n hu\u1ef3nh quang b\u1eb1ng c\u00e1ch s\u1eed d\u1ee5ng \u00e1nh s\u00e1ng truy\u1ec1n qua, sau \u0111\u00f3 l\u00e0 h\u00ecnh \u1ea3nh \u1edf ch\u1ebf \u0111\u1ed9 hu\u1ef3nh quang epi.\u00a0Hai h\u00ecnh \u1ea3nh c\u00f3 th\u1ec3 \u0111\u01b0\u1ee3c k\u1ebft h\u1ee3p trong qu\u00e1 tr\u00ecnh ph\u00e2n t\u00edch.\u00a0C\u00e1c c\u1ea5u h\u00ecnh k\u00ednh hi\u1ec3n vi tr\u01b0\u1eddng r\u1ed9ng ti\u00ean ti\u1ebfn c\u00f3 th\u1ec3 ch\u1ee5p \u0111\u1ed3ng th\u1eddi h\u00ecnh \u1ea3nh DIC v\u00e0 hu\u1ef3nh quang b\u1eb1ng c\u00e1ch s\u1eed d\u1ee5ng h\u1ec7 th\u1ed1ng chi\u1ebfu s\u00e1ng \u0111\u01b0\u1ee3c ph\u00e2n t\u00e1ch b\u1eb1ng quang ph\u1ed5 (nh\u01b0 kh\u1ea3 ki\u1ebfn \u200b\u200bv\u00e0 c\u1eadn h\u1ed3ng ngo\u1ea1i) v\u00e0 m\u00e1y \u1ea3nh k\u00e9p ho\u1eb7c b\u1ed9 \u0111i\u1ec1u h\u1ee3p m\u00e1y \u1ea3nh c\u00f3 ch\u1ebf \u0111\u1ed9 xem t\u00e1ch.\u00a0Trong h\u1ea7u h\u1ebft c\u00e1c k\u00ednh hi\u1ec3n vi \u0111\u1ed3ng ti\u00eau qu\u00e9t laze (th\u1ea3o lu\u1eadn b\u00ean d\u01b0\u1edbi), h\u00ecnh \u1ea3nh c\u00f3 th\u1ec3 \u0111\u01b0\u1ee3c thu nh\u1eadn \u0111\u1ed3ng th\u1eddi \u1edf ch\u1ebf \u0111\u1ed9 hu\u1ef3nh quang v\u00e0 DIC, do \u0111\u00f3 lo\u1ea1i b\u1ecf nhu c\u1ea7u x\u1eed l\u00fd sau thu nh\u1eadn.\u00a0\u0110\u1ed9 t\u01b0\u01a1ng ph\u1ea3n pha v\u00e0 \u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n \u0111i\u1ec1u ch\u1ebf Hoffman (H\u00ecnh 5(d) v\u00e0 5(e)) c\u0169ng c\u00f3 th\u1ec3 \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng th\u00e0nh c\u00f4ng khi k\u1ebft h\u1ee3p v\u1edbi k\u00ednh hi\u1ec3n vi hu\u1ef3nh quang tr\u01b0\u1eddng r\u1ed9ng.\u00a0Tuy nhi\u00ean, trong c\u00e1c k\u1ef9 thu\u1eadt n\u00e0y, c\u00e1c v\u1eadt k\u00ednh \u0111\u01b0\u1ee3c s\u1eeda \u0111\u1ed5i tr\u1ef1c ti\u1ebfp, b\u1eb1ng v\u00f2ng pha ho\u1eb7c t\u1ea5m \u0111i\u1ec1u ch\u1ebf Hoffman, \u0111i\u1ec1u n\u00e0y s\u1ebd d\u1eabn \u0111\u1ebfn m\u1ea5t c\u01b0\u1eddng \u0111\u1ed9 ph\u00e1t x\u1ea1 l\u00ean t\u1edbi 5-15 ph\u1ea7n tr\u0103m.<\/p>\n<p>M\u1ed9t l\u01b0u \u00fd l\u00e0 khi k\u1ebft h\u1ee3p DIC, \u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n pha ho\u1eb7c \u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n \u0111i\u1ec1u ch\u1ebf Hoffman v\u1edbi hu\u1ef3nh quang, \u0111\u1eb7c bi\u1ec7t l\u00e0 trong ch\u1ee5p \u1ea3nh t\u1ebf b\u00e0o s\u1ed1ng, \u00e1nh s\u00e1ng ph\u00e1t x\u1ea1 truy\u1ec1n qua m\u00e1y ph\u00e2n t\u00edch DIC, v\u00f2ng pha v\u1eadt k\u00ednh ho\u1eb7c t\u1ea5m \u0111i\u1ec1u ch\u1ebf Hoffman \u1edf ph\u00eda sau s\u1ebd b\u1ecb m\u1ea5t \u0111i. Trong tr\u01b0\u1eddng h\u1ee3p tr\u01b0\u1edbc, m\u00e1y ph\u00e2n t\u00edch DIC v\u00e0 l\u0103ng k\u00ednh Nomarski ph\u1ea3i \u0111\u01b0\u1ee3c lo\u1ea1i b\u1ecf kh\u1ecfi \u0111\u01b0\u1eddng ch\u00f9m tia tr\u01b0\u1edbc khi ch\u1ee5p \u1ea3nh hu\u1ef3nh quang, nh\u01b0ng v\u1eadt k\u00ednh pha v\u00e0 v\u1eadt k\u00ednh Hoffman ch\u1ee9a c\u00e1c th\u00e0nh ph\u1ea7n quang \u0111\u00e3 s\u1eeda \u0111\u1ed5i kh\u00f4ng th\u1ec3 lo\u1ea1i b\u1ecf \u0111\u01b0\u1ee3c. S\u1ef1 m\u1ea5t \u00e1nh s\u00e1ng truy\u1ec1n qua v\u00f2ng pha ho\u1eb7c b\u1ed9 bi\u1ebfn \u0111i\u1ec7u Hoffman kh\u00f4ng \u0111\u00e1ng k\u1ec3 b\u1eb1ng s\u1ef1 m\u1ea5t \u00e1nh s\u00e1ng qua b\u1ed9 ph\u00e2n c\u1ef1c. Tuy nhi\u00ean, c\u00e1c m\u1eabu v\u1eadt \u0111\u01b0\u1ee3c nhu\u1ed9m hu\u1ef3nh quang k\u00e9m, vi\u1ec7c t\u0103ng \u0111\u1ed9 nh\u1ea1y b\u1eb1ng c\u00e1ch s\u1eed d\u1ee5ng c\u00e1c v\u1eadt k\u00ednh c\u00f3 \u0111\u1ed9 truy\u1ec1n s\u00e1ng cao l\u00e0 r\u1ea5t quan tr\u1ecdng<\/p>\n<p>Minh h\u1ecda trong <em>H\u00ecnh 5<\/em> l\u00e0 m\u1ed9t lo\u1ea1t \u1ea3nh \u0111\u01b0\u1ee3c ch\u1ee5p b\u1eb1ng k\u00ednh hi\u1ec3n vi hu\u1ef3nh quang tr\u01b0\u1eddng r\u1ed9ng, ho\u1eb7c k\u1ebft h\u1ee3p v\u1edbi m\u1ed9t s\u1ed1 k\u1ef9 thu\u1eadt \u00e1nh s\u00e1ng truy\u1ec1n qua \u0111\u01b0\u1ee3c m\u00f4 t\u1ea3 \u1edf tr\u00ean. C\u00e1c t\u1ebf b\u00e0o ung th\u01b0 bi\u1ec3u m\u00f4 ng\u01b0\u1eddi s\u1ed1ng (d\u00f2ng <b>HeLa<\/b>) th\u1ec3 hi\u1ec7n s\u1ef1 k\u1ebft h\u1ee3p c\u1ee7a protein hu\u1ef3nh quang m\u00e0u xanh l\u00e1 c\u00e2y t\u0103ng c\u01b0\u1eddng (<b>EGFP<\/b>) h\u1ee3p nh\u1ea5t v\u1edbi cytokeratin c\u1ee7a con ng\u01b0\u1eddi \u0111\u01b0\u1ee3c th\u1ec3 hi\u1ec7n trong <em>H\u00ecnh 5(a)<\/em>.\u00a0C\u00e1c t\u1ebf b\u00e0o HeLa k\u1ebft d\u00ednh c\u00f3 \u0111\u1ed9 d\u00e0y kho\u1ea3ng 3 \u0111\u1ebfn 5 micromet v\u00e0 lu\u00f4n \u0111\u01b0\u1ee3c l\u1ea5y n\u00e9t trong su\u1ed1t h\u00ecnh \u1ea3nh.\u00a0Ng\u01b0\u1ee3c l\u1ea1i, c\u00e1c m\u1eabu m\u00f4 d\u00e0y (d\u1ea1 d\u00e0y chu\u1ed9t; <em>H\u00ecnh 5(b)<\/em>) r\u1ea5t kh\u00f3 ph\u00e2n gi\u1ea3i trong hu\u1ef3nh quang tr\u01b0\u1eddng r\u1ed9ng do s\u1ef1 \u0111\u00f3ng g\u00f3p c\u1ee7a c\u00e1c c\u1ea5u tr\u00fac hu\u1ef3nh quang kh\u00f4ng n\u1eb1m trong m\u1eb7t ph\u1eb3ng ti\u00eau c\u1ef1 c\u1ee7a k\u00ednh hi\u1ec3n vi.\u00a0Hu\u1ef3nh quang tr\u01b0\u1eddng r\u1ed9ng c\u00f3 th\u1ec3 \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng k\u1ebft h\u1ee3p v\u1edbi nhi\u1ec1u k\u1ef9 thu\u1eadt \u00e1nh s\u00e1ng truy\u1ec1n qua, bao g\u1ed3m DIC (ph\u1ea7n m\u1ecfng th\u1eadn chu\u1ed9t; <em>H\u00ecnh 5(c)<\/em>), t\u01b0\u01a1ng ph\u1ea3n \u0111i\u1ec1u ch\u1ebf Hoffman (t\u1ebf b\u00e0o chu\u1ed9t ph\u00f4i; <em>H\u00ecnh 5(d)<\/em>) v\u00e0 t\u01b0\u01a1ng ph\u1ea3n pha (th\u1ecf t\u1ebf b\u00e0o th\u1eadn; <em>H\u00ecnh 5(e)<\/em>).\u00a0S\u1eed d\u1ee5ng k\u1ebft h\u1ee3p c\u00e1c b\u1ed9 l\u1ecdc nhi\u1ec5u (t\u1ebf b\u00e0o Muntjac c\u1ee7a \u1ea4n \u0110\u1ed9; <em>H\u00ecnh 5(f)<\/em>).<\/p>\n<h2><span class=\"ez-toc-section\" id=\"Kinh_hien_vi_dong_tieu_quet_laze\"><\/span><strong><a href=\"https:\/\/vospan.com\/product-brands\/zeiss-vi\/\">K\u00ednh hi\u1ec3n vi \u0111\u1ed3ng ti\u00eau<\/a> qu\u00e9t laze<\/strong><span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p>K\u00ednh hi\u1ec3n vi \u0111\u1ed3ng ti\u00eau qu\u00e9t laze mang l\u1ea1i m\u1ed9t s\u1ed1 \u01b0u \u0111i\u1ec3m so v\u1edbi k\u00ednh hi\u1ec3n vi hu\u1ef3nh quang tr\u01b0\u1eddng r\u1ed9ng th\u00f4ng th\u01b0\u1eddng, bao g\u1ed3m \u0111\u1ed9 s\u00e2u tr\u01b0\u1eddng \u1ea3nh c\u00f3 th\u1ec3 ki\u1ec3m so\u00e1t, lo\u1ea1i b\u1ecf th\u00f4ng tin l\u00e0m gi\u1ea3m \u0111\u1ed9 n\u00e9t c\u1ee7a h\u00ecnh \u1ea3nh v\u00e0 kh\u1ea3 n\u0103ng thu th\u1eadp\u00a0<b>c\u00e1c ph\u1ea7n quang h\u1ecdc n\u1ed1i ti\u1ebfp <\/b>t\u1eeb m\u1eabu d\u00e0y.\u00a0Ch\u00eca kh\u00f3a c\u1ee7a ph\u01b0\u01a1ng ph\u00e1p ti\u1ebfp c\u1eadn \u0111\u1ed3ng ti\u00eau l\u00e0 s\u1eed d\u1ee5ng b\u1ed9 l\u1ecdc kh\u00f4ng gian \u0111\u1ec3 lo\u1ea1i b\u1ecf \u00e1nh s\u00e1ng ngo\u00e0i ti\u00eau c\u1ef1 ho\u1eb7c l\u00f3a trong c\u00e1c m\u1eabu v\u1eadt d\u00e0y h\u01a1n m\u1eb7t ph\u1eb3ng ti\u00eau \u0111i\u1ec3m b\u1eb1ng c\u00e1ch chi\u1ebfu s\u00e1ng v\u1eadt k\u00ednh qua l\u1ed7 kim (xem <em>H\u00ecnh 6<\/em> v\u00e0 <em>H\u00ecnh 8(b)<\/em>) .\u00a0H\u00ecnh \u1ea3nh c\u1ee7a l\u1ed7 kim \u1edf d\u1ea1ng m\u1ed9t \u0111i\u1ec3m nh\u1ecf \u0111\u01b0\u1ee3c h\u00ecnh th\u00e0nh tr\u00ean m\u1eabu v\u1eadt b\u1eb1ng tia laser h\u1ed9i t\u1ee5 \u0111\u01b0\u1ee3c \u0111i\u1ec1u khi\u1ec3n b\u1edfi h\u1ec7 th\u1ed1ng qu\u00e9t d\u1ef1a tr\u00ean \u0111i\u1ec7n k\u1ebf.\u00a0\u0110\u1ebfn l\u01b0\u1ee3t n\u00f3, \u0111i\u1ec3m n\u00e0y t\u1ea1o th\u00e0nh h\u00ecnh \u1ea3nh hu\u1ef3nh quang epi ph\u1ea3n x\u1ea1 tr\u00ean l\u1ed7 kim ban \u0111\u1ea7u.\u00a0N\u1ebfu m\u1eabu v\u1eadt \u0111\u01b0\u1ee3c l\u1ea5y n\u00e9t, \u00e1nh s\u00e1ng s\u1ebd \u0111i qua l\u1ed7 kim \u0111\u1ebfn m\u00e1y d\u00f2 (th\u01b0\u1eddng l\u00e0 b\u1ed9 nh\u00e2n quang).\u00a0Khi m\u1eabu v\u1eadt kh\u00f4ng \u0111\u01b0\u1ee3c l\u1ea5y n\u00e9t, \u00e1nh s\u00e1ng ph\u1ea3n x\u1ea1 t\u1eeb n\u00f3 b\u1ecb m\u1ea5t t\u1eadp trung t\u1ea1i l\u1ed7 kim v\u00e0 r\u1ea5t \u00edt \u0111i qua.\u00a0Do \u0111\u00f3, ph\u00e1t x\u1ea1 hu\u1ef3nh quang quay tr\u1edf l\u1ea1i t\u1eeb m\u1eabu v\u1eadt \u0111\u1ebfn m\u00e1y d\u00f2 \u0111\u01b0\u1ee3c l\u1ecdc theo kh\u00f4ng gian.\u00a0Khi kh\u1ea9u \u0111\u1ed9 l\u1ed7 kim \u0111\u01b0\u1ee3c gi\u1ea3m k\u00edch th\u01b0\u1edbc, n\u00f3 s\u1ebd ch\u1eb7n \u0111\u01b0\u1ee3c nhi\u1ec1u \u00e1nh s\u00e1ng \u0111i l\u1ea1c h\u01a1n nh\u01b0ng c\u0169ng l\u00e0m gi\u1ea3m m\u1ee9c t\u00edn hi\u1ec7u t\u1ed5ng.\u00a0M\u1eb7c d\u00f9 gi\u00e1 tr\u1ecb t\u00edn hi\u1ec7u tuy\u1ec7t \u0111\u1ed1i th\u1ea5p h\u01a1n nhi\u1ec1u so v\u1edbi quan s\u00e1t \u0111\u01b0\u1ee3c v\u1edbi c\u1ea5u h\u00ecnh k\u00ednh hi\u1ec3n vi tr\u01b0\u1eddng r\u1ed9ng, nh\u01b0ng vi\u1ec7c lo\u1ea1i b\u1ecf \u00e1nh s\u00e1ng t\u1eeb c\u00e1c m\u1eb7t ph\u1eb3ng ti\u00eau c\u1ef1 kh\u00e1c s\u1ebd l\u00e0m t\u0103ng t\u1ef7 l\u1ec7 t\u00edn hi\u1ec7u tr\u00ean t\u1ea1p \u00e2m c\u1ee5 th\u1ec3 cho c\u00e1c t\u00ednh n\u0103ng quan t\u00e2m.<\/p>\n<p>C\u00e1c \u01b0u \u0111i\u1ec3m kh\u00e1c c\u1ee7a k\u00ednh hi\u1ec3n vi qu\u00e9t \u0111\u1ed3ng ti\u00eau qu\u00e9t laze bao g\u1ed3m kh\u1ea3 n\u0103ng \u0111i\u1ec1u ch\u1ec9nh \u0111\u1ed9 ph\u00f3ng \u0111\u1ea1i b\u1eb1ng \u0111i\u1ec7n t\u1eed b\u1eb1ng c\u00e1ch thay \u0111\u1ed5i khu v\u1ef1c \u0111\u01b0\u1ee3c qu\u00e9t b\u1edfi tia laze m\u00e0 kh\u00f4ng ph\u1ea3i thay \u0111\u1ed5i v\u1eadt k\u00ednh.\u00a0T\u00ednh n\u0103ng n\u00e0y \u0111\u01b0\u1ee3c g\u1ecdi l\u00e0\u00a0<b>zoom <\/b>v\u00e0 th\u01b0\u1eddng \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng \u0111\u1ec3 \u0111i\u1ec1u ch\u1ec9nh \u0111\u1ed9 ph\u00e2n gi\u1ea3i kh\u00f4ng gian c\u1ee7a h\u00ecnh \u1ea3nh b\u1eb1ng c\u00e1ch thay \u0111\u1ed5i chu k\u1ef3 l\u1ea5y m\u1eabu laze qu\u00e9t. Vi\u1ec7c t\u0103ng h\u1ec7 s\u1ed1 thu ph\u00f3ng s\u1ebd l\u00e0m gi\u1ea3m di\u1ec7n t\u00edch m\u1eabu \u0111\u01b0\u1ee3c qu\u00e9t v\u00e0 \u0111\u1ed3ng th\u1eddi gi\u1ea3m t\u1ed1c \u0111\u1ed9 qu\u00e9t. K\u1ebft qu\u1ea3 l\u00e0 s\u1ed1 l\u01b0\u1ee3ng m\u1eabu t\u0103ng l\u00ean d\u1ecdc theo chi\u1ec1u d\u00e0i t\u01b0\u01a1ng \u0111\u01b0\u01a1ng, gi\u00fap t\u0103ng c\u1ea3 \u0111\u1ed9 ph\u00e2n gi\u1ea3i kh\u00f4ng gian h\u00ecnh \u1ea3nh v\u00e0 \u0111\u1ed9 ph\u00f3ng \u0111\u1ea1i hi\u1ec3n th\u1ecb tr\u00ean m\u00e0n h\u00ecnh m\u00e1y t\u00ednh ch\u1ee7. Thu ph\u00f3ng \u0111\u1ed3ng ti\u00eau th\u01b0\u1eddng \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng \u0111\u1ec3 kh\u1edbp \u0111\u1ed9 ph\u00e2n gi\u1ea3i h\u00ecnh \u1ea3nh k\u1ef9 thu\u1eadt s\u1ed1 v\u1edbi \u0111\u1ed9 ph\u00e2n gi\u1ea3i quang h\u1ecdc c\u1ee7a k\u00ednh hi\u1ec3n vi khi c\u00e1c v\u1eadt k\u00ednh c\u00f3 kh\u1ea9u \u0111\u1ed9 v\u00e0 \u0111\u1ed9 ph\u00f3ng \u0111\u1ea1i s\u1ed1 th\u1ea5p \u0111ang \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng \u0111\u1ec3 thu th\u1eadp d\u1eef li\u1ec7u. \u0110i\u1ec1u khi\u1ec3n thu ph\u00f3ng n\u00ean \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng m\u1ed9t c\u00e1ch th\u1eadn tr\u1ecdng do th\u1ef1c t\u1ebf l\u00e0 c\u00e1c h\u1ec7 s\u1ed1 thu ph\u00f3ng cao d\u1eabn \u0111\u1ebfn t\u0103ng t\u1ea9y tr\u1eafng quang.<\/p>\n<p><img loading=\"lazy\" decoding=\"async\" class=\"aligncenter\" src=\"https:\/\/zeiss-campus.magnet.fsu.edu\/articles\/livecellimaging\/images\/techniquesfigure6.jpg\" width=\"392\" height=\"327\" border=\"0\" \/>Nh\u01b0\u1ee3c \u0111i\u1ec3m ch\u00ednh c\u1ee7a k\u00ednh hi\u1ec3n vi \u0111\u1ed3ng ti\u00eau qu\u00e9t laser th\u00f4ng th\u01b0\u1eddng \u0111\u1ec3 ch\u1ee5p \u1ea3nh t\u1ebf b\u00e0o v\u00e0 m\u00f4 l\u00e0 h\u00ecnh \u1ea3nh \u0111\u01b0\u1ee3c thu th\u1eadp b\u1eb1ng c\u00e1ch qu\u00e9t raster m\u1eabu v\u1eadt, qu\u00e1 tr\u00ecnh n\u00e0y t\u01b0\u01a1ng \u0111\u1ed1i ch\u1eadm v\u00e0 th\u01b0\u1eddng c\u1ea7n v\u00e0i gi\u00e2y ho\u1eb7c l\u00e2u h\u01a1n cho m\u1ed7i h\u00ecnh \u1ea3nh.\u00a0Ngo\u00e0i ra c\u00f2n c\u00f3 m\u1ed9t m\u1ee9c \u0111\u1ed9 r\u1ee7i ro cao t\u1eeb vi\u1ec7c chi\u1ebfu x\u1ea1 m\u1eabu v\u1eadt b\u1eb1ng \u00e1nh s\u00e1ng laze c\u01b0\u1eddng \u0111\u1ed9 cao c\u00f3 th\u1ec3 t\u1ea1o ra \u0111\u1ed9c t\u1ed1 quang trong c\u00e1c t\u1ebf b\u00e0o s\u1ed1ng.\u00a0Trong h\u1ea7u h\u1ebft c\u00e1c tr\u01b0\u1eddng h\u1ee3p, th\u1eddi gian d\u1eebng c\u1ee7a ch\u00f9m tia laze t\u1ea1i b\u1ea5t k\u1ef3 v\u1ecb tr\u00ed n\u00e0o tr\u00ean m\u1eabu v\u1eadt ch\u1ec9 l\u00e0 v\u00e0i micro gi\u00e2y.\u00a0Do \u0111\u00f3, n\u0103ng l\u01b0\u1ee3ng k\u00edch th\u00edch b\u1eb1ng laser ph\u1ea3i \u0111\u1ee7 cao \u0111\u1ec3 t\u1ea1o ra t\u00edn hi\u1ec7u c\u00f3 th\u1ec3 s\u1eed d\u1ee5ng \u0111\u01b0\u1ee3c trong th\u1eddi gian d\u1eebng \u0111\u00f3, th\u01b0\u1eddng d\u1eabn \u0111\u1ebfn s\u1ef1 b\u00e3o h\u00f2a ho\u00e0n to\u00e0n c\u1ee7a t\u1ea5t c\u1ea3 c\u00e1c ch\u1ea5t hu\u1ef3nh quang c\u01b0 tr\u00fa t\u1ea1i \u0111i\u1ec3m.\u00a0Trong nh\u1eefng \u0111i\u1ec1u ki\u1ec7n n\u00e0y, qu\u00e1 tr\u00ecnh t\u1ea9y tr\u1eafng quang di\u1ec5n ra nhanh ch\u00f3ng v\u00e0 vi\u1ec7c thu th\u1eadp h\u00ecnh \u1ea3nh c\u00f3 d\u1ea3i \u0111\u1ed9ng n\u1ed9i c\u1ea3nh cao (m\u1ed9t s\u1ed1 l\u01b0\u1ee3ng \u0111\u00e1ng k\u1ec3 c\u00e1c m\u1ee9c x\u00e1m) l\u00e0 m\u1ed9t th\u00e1ch th\u1ee9c.\u00a0\u0110\u1ed1i v\u1edbi h\u00ecnh \u1ea3nh t\u1ebf b\u00e0o s\u1ed1ng trong th\u1eddi gian d\u00e0i, n\u00ean gi\u1ea3m c\u01b0\u1eddng \u0111\u1ed9 laser c\u00e0ng nhi\u1ec1u c\u00e0ng t\u1ed1t.\u00a0Th\u00e1ch th\u1ee9c l\u1edbn \u0111\u1ed1i v\u1edbi vi\u1ec7c ch\u1ee5p \u1ea3nh t\u1ebf b\u00e0o s\u1ed1ng b\u1eb1ng c\u00e1ch s\u1eed d\u1ee5ng k\u00ednh hi\u1ec3n vi \u0111\u1ed3ng ti\u00eau qu\u00e9t laser l\u00e0 t\u1ea1o ra \u0111\u1ee7 \u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n trong khi gi\u1ea3m \u0111\u1ed9c t\u00ednh quang, \u0111\u1eb7c bi\u1ec7t l\u00e0 trong tr\u01b0\u1eddng h\u1ee3p c\u00e1c t\u1ebf b\u00e0o \u0111\u01b0\u1ee3c ch\u1ee5p \u1ea3nh trong th\u1eddi gian d\u00e0i.<\/p>\n<h2 class=\"paratitle\"><span class=\"ez-toc-section\" id=\"Kinh_hien_vi_dong_tieu_dia_quay\"><\/span><strong><a href=\"https:\/\/vospan.com\/product-brands\/zeiss-vi\/\">K\u00ednh hi\u1ec3n vi \u0111\u1ed3ng ti\u00eau<\/a> \u0111\u0129a quay<\/strong><span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p>M\u1ed9t bi\u1ebfn th\u1ec3 c\u1ee7a k\u00ednh hi\u1ec3n vi \u0111\u1ed3ng ti\u00eau d\u1ef1a tr\u00ean \u0111i\u1ec7n k\u1ebf l\u00e0<b>\u00a0<\/b>k\u00ednh hi\u1ec3n vi \u0111\u1ed3ng ti\u00eau <strong>\u0111\u0129a quay<\/strong>, c\u00f3 th\u1ec3 ho\u1ea1t \u0111\u1ed9ng trong th\u1eddi gian th\u1ef1c (30 khung h\u00ecnh m\u1ed7i gi\u00e2y) ho\u1eb7c th\u1eadm ch\u00ed nhanh h\u01a1n \u0111\u1ec3 n\u1eafm b\u1eaft c\u00e1c s\u1ef1 ki\u1ec7n \u0111\u1ed9ng trong m\u1ed9t d\u1ea3i th\u1eddi gian r\u1ed9ng. Nguy\u00ean t\u1eafc n\u00e0y d\u1ef1a tr\u00ean m\u1ed9t \u0111\u0129a ki\u1ec3u Nipkow kh\u00f4ng trong su\u1ed1t, ngo\u1ea1i tr\u1eeb h\u00e0ng ngh\u00ecn l\u1ed7 kim \u0111\u01b0\u1ee3c khoan ho\u1eb7c kh\u1eafc, th\u01b0\u1eddng \u0111\u01b0\u1ee3c ph\u1ee7 b\u1eb1ng th\u1ea5u k\u00ednh h\u1ed9i t\u1ee5 thu nh\u1ecf, \u0111\u01b0\u1ee3c s\u1eafp x\u1ebfp theo m\u00f4 h\u00ecnh xo\u1eafn \u1ed1c Archimedean xen k\u1ebd. M\u1ed7i l\u1ed7 kim \u0111\u01b0\u1ee3c chi\u1ebfu s\u00e1ng tr\u00ean \u0111\u0129a \u0111\u01b0\u1ee3c v\u1eadt k\u00ednh t\u1ea1o \u1ea3nh th\u00e0nh m\u1ed9t \u0111i\u1ec3m gi\u1edbi h\u1ea1n nhi\u1ec5u x\u1ea1 tr\u00ean m\u1eabu v\u1eadt. Ph\u00e1t x\u1ea1 hu\u1ef3nh quang ph\u1ea3n x\u1ea1 t\u1eeb m\u1eabu v\u1eadt c\u00f3 th\u1ec3 \u0111\u01b0\u1ee3c quan s\u00e1t v\u00e0 ghi l\u1ea1i sau khi n\u00f3 \u0111i qua c\u00e1c l\u1ed7 kim c\u1ee7a \u0111\u0129a Nipkow. V\u00e0i ngh\u00ecn \u0111i\u1ec3m tr\u00ean m\u1eabu v\u1eadt \u0111\u01b0\u1ee3c chi\u1ebfu s\u00e1ng \u0111\u1ed3ng th\u1eddi b\u1edfi \u0111\u0129a, tr\u00ean th\u1ef1c t\u1ebf, \u0111\u1ea1t \u0111\u01b0\u1ee3c v\u00e0i ngh\u00ecn k\u00ednh hi\u1ec3n vi \u0111\u1ed3ng ti\u00eau, t\u1ea5t c\u1ea3 \u0111\u1ec1u ch\u1ea1y song song.<\/p>\n<p>M\u1ed9t gi\u1ea3i ph\u00e1p thay th\u1ebf cho \u0111\u0129a Nipkow li\u00ean quan \u0111\u1ebfn c\u00f4ng ngh\u1ec7 g\u1ea7n \u0111\u00e2y cung c\u1ea5p th\u00f4ng l\u01b0\u1ee3ng \u00e1nh s\u00e1ng cao b\u1eb1ng c\u00e1ch s\u1eed d\u1ee5ng \u0111\u0129a \u0111\u01b0\u1ee3c kh\u1eafc b\u1eb1ng m\u00f4 h\u00ecnh c\u00e1c khe vu\u00f4ng g\u00f3c, thay v\u00ec l\u1ed7 kim, mang l\u1ea1i kh\u1ea3 n\u0103ng truy\u1ec1n d\u1eabn l\u1edbn h\u01a1n trong khi v\u1eabn duy tr\u00ec m\u1ee9c \u0111\u1ed9 \u0111\u1ed3ng ti\u00eau c\u00f3 th\u1ec3 ch\u1ea5p nh\u1eadn \u0111\u01b0\u1ee3c (<em>H\u00ecnh 7<\/em>).\u00a0Kh\u00f4ng gi\u1ed1ng nh\u01b0 c\u00e1c \u0111\u0129a ki\u1ec3u Nipkow, c\u00e1c \u0111\u0129a ki\u1ec3u khe n\u00e0y \u0111\u01b0\u1ee3c s\u1ea3n xu\u1ea5t v\u1edbi c\u00e1c \u0111\u1ed9 r\u1ed9ng khe kh\u00e1c nhau ph\u00f9 h\u1ee3p v\u1edbi c\u00e1c kh\u1ea9u \u0111\u1ed9, \u0111\u1ed9 ph\u00f3ng \u0111\u1ea1i v\u00e0 \u0111\u1ed9 d\u00e0y m\u1eabu v\u1eadt kh\u00e1c nhau.\u00a0Do \u0111\u00f3, c\u00e1c \u0111\u0129a khe cho ph\u00e9p thu \u0111\u01b0\u1ee3c h\u00ecnh \u1ea3nh \u0111\u1ed3ng ti\u00eau \u1edf \u0111\u1ed9 ph\u00e2n gi\u1ea3i cao b\u1eb1ng c\u00e1ch s\u1eed d\u1ee5ng c\u00e1c v\u1eadt k\u00ednh c\u00f3 \u0111\u1ed9 ph\u00f3ng \u0111\u1ea1i t\u1eeb 10x \u0111\u1ebfn 100x.\u00a0C\u1ea3 hai thi\u1ebft k\u1ebf \u0111\u0129a quay \u0111\u1ec1u \u0111\u01b0\u1ee3c ph\u00e1t tri\u1ec3n ch\u1ee7 y\u1ebfu cho c\u00e1c \u1ee9ng d\u1ee5ng t\u1ea1o \u1ea3nh t\u1ebf b\u00e0o s\u1ed1ng, n\u01a1i t\u1ed3n t\u1ea1i s\u1ef1 th\u1ecfa hi\u1ec7p gi\u1eefa vi\u1ec7c t\u0103ng t\u1ed1c \u0111\u1ed9 thu nh\u1eadn \u1ea3nh c\u1ea7n thi\u1ebft v\u1edbi vi\u1ec7c gi\u1ea3m nh\u1eb9 \u0111\u1ed9 ph\u00e2n gi\u1ea3i tr\u1ee5c (<em>H\u00ecnh 8(c)<\/em>).<\/p>\n<p><img loading=\"lazy\" decoding=\"async\" class=\"aligncenter\" src=\"https:\/\/zeiss-campus.magnet.fsu.edu\/articles\/livecellimaging\/images\/techniquesfigure7.jpg\" alt=\"K\u00ednh hi\u1ec3n vi\" width=\"482\" height=\"395\" border=\"0\" \/>M\u1ed9t s\u1ed1 nh\u00e0 s\u1ea3n xu\u1ea5t \u0111\u00e3 gi\u1edbi thi\u1ec7u c\u00e1c h\u1ec7 th\u1ed1ng ti\u00eau \u0111i\u1ec3m s\u1eed d\u1ee5ng \u00e1nh s\u00e1ng laze \u0111\u01b0\u1ee3c \u0111\u1ecbnh h\u00ecnh th\u00e0nh m\u1ed9t \u0111\u01b0\u1eddng ch\u1ee9 kh\u00f4ng ph\u1ea3i l\u00e0 m\u1ed9t \u0111i\u1ec3m \u0111\u1ec3 qu\u00e9t m\u1eabu v\u1eadt, nh\u01b0ng gi\u1ea3m \u0111\u00e1ng k\u1ec3 th\u1eddi gian thu nh\u1eadn h\u00ecnh \u1ea3nh do \u0111\u1ed9 ph\u00e2n gi\u1ea3i (th\u01b0\u1eddng l\u00e0 h\u1ec7 s\u1ed1 1,2 trong m\u1ed9t chi\u1ec1u).\u00a0Kh\u00f4ng gi\u1ed1ng nh\u01b0 tr\u01b0\u1eddng h\u1ee3p v\u1edbi k\u00ednh hi\u1ec3n vi \u0111\u0129a quay, m\u1ee9c \u0111\u1ed9 \u0111\u1ed3ng ti\u00eau c\u00f3 th\u1ec3 \u0111\u01b0\u1ee3c ki\u1ec3m so\u00e1t b\u1edfi ng\u01b0\u1eddi d\u00f9ng b\u1eb1ng c\u00e1ch g\u1ecdi c\u00e1c thay \u0111\u1ed5i \u0111\u1ed1i v\u1edbi h\u00ecnh d\u1ea1ng v\u00e0 k\u00edch th\u01b0\u1edbc c\u1ee7a \u0111\u01b0\u1eddng.\u00a0K\u00ednh hi\u1ec3n vi qu\u00e9t d\u00f2ng \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng v\u1edbi h\u1ec7 th\u1ed1ng camera CCD ch\u1ee9 kh\u00f4ng ph\u1ea3i l\u00e0 b\u1ed9 nh\u00e2n quang v\u00e0 th\u01b0\u1eddng c\u00f3 t\u1ed1c \u0111\u1ed9 thu nh\u1eadn nhanh h\u01a1n nhi\u1ec1u so v\u1edbi \u0111\u0129a quay ho\u1eb7c k\u00ednh hi\u1ec3n vi \u0111\u1ed3ng ti\u00eau qu\u00e9t laze.\u00a0T\u00f3m l\u1ea1i, \u0111\u00e3 c\u00f3 m\u1ed9t s\u1ef1 b\u00f9ng n\u1ed5 to l\u1edbn v\u1ec1 m\u1ee9c \u0111\u1ed9 ph\u1ed5 bi\u1ebfn c\u1ee7a k\u00ednh hi\u1ec3n vi \u0111\u1ed3ng ti\u00eau trong nh\u1eefng n\u0103m g\u1ea7n \u0111\u00e2y,<\/p>\n<h2 class=\"paratitle\"><span class=\"ez-toc-section\" id=\"Kinh_hien_vi_da_diem\"><\/span><strong><a href=\"https:\/\/vospan.com\/product-brands\/zeiss-vi\/\">K\u00ednh hi\u1ec3n vi<\/a> \u0111a \u0111i\u1ec3m<\/strong><span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p>S\u1ef1 k\u00edch th\u00edch trong k\u00ednh hi\u1ec3n vi \u0111a photon l\u00e0 m\u1ed9t qu\u00e1 tr\u00ecnh phi tuy\u1ebfn t\u00ednh ch\u1ec9 x\u1ea3y ra t\u1ea1i ti\u00eau \u0111i\u1ec3m c\u1ee7a k\u00ednh hi\u1ec3n vi gi\u1edbi h\u1ea1n nhi\u1ec5u x\u1ea1, cung c\u1ea5p kh\u1ea3 n\u0103ng c\u1eaft c\u00e1c m\u1eabu v\u1eadt sinh h\u1ecdc d\u00e0y v\u1ec1 m\u1eb7t quang h\u1ecdc \u0111\u1ec3 thu \u0111\u01b0\u1ee3c \u0111\u1ed9 ph\u00e2n gi\u1ea3i ba chi\u1ec1u.\u00a0C\u00e1c ph\u1ea7n quang h\u1ecdc ri\u00eang l\u1ebb c\u00f3 \u0111\u01b0\u1ee3c b\u1eb1ng c\u00e1ch qu\u00e9t raster m\u1eabu v\u1eadt trong m\u1eb7t ph\u1eb3ng\u00a0<b>x<\/b>\u00a0&#8211;\u00a0<b>y<\/b> v\u00e0 h\u00ecnh \u1ea3nh ba chi\u1ec1u \u0111\u1ea7y \u0111\u1ee7 \u0111\u01b0\u1ee3c t\u1ea1o b\u1eb1ng c\u00e1ch qu\u00e9t li\u00ean ti\u1ebfp m\u1eabu v\u1eadt theo tr\u1ee5c li\u00ean ti\u1ebfp (<b>z<\/b>) c\u00e1c v\u1ecb tr\u00ed, t\u01b0\u01a1ng t\u1ef1 nh\u01b0 tr\u01b0\u1eddng h\u1ee3p trong k\u00ednh hi\u1ec3n vi \u0111\u1ed3ng ti\u00eau (\u0111\u1ec3 t\u1ea1o ra c\u00e1c ph\u1ea7n quang h\u1ecdc).\u00a0Do v\u1ecb tr\u00ed c\u1ee7a ti\u00eau \u0111i\u1ec3m c\u00f3 th\u1ec3 \u0111\u01b0\u1ee3c x\u00e1c \u0111\u1ecbnh v\u00e0 ki\u1ec3m so\u00e1t ch\u00ednh x\u00e1c n\u00ean hu\u1ef3nh quang \u0111a photon \u0111\u1eb7c bi\u1ec7t h\u1eefu \u00edch \u0111\u1ec3 th\u0103m d\u00f2 c\u00e1c v\u00f9ng \u0111\u01b0\u1ee3c ch\u1ecdn b\u00ean d\u01b0\u1edbi b\u1ec1 m\u1eb7t m\u1eabu v\u1eadt.\u00a0N\u0103ng l\u01b0\u1ee3ng k\u00edch th\u00edch c\u1ee5c b\u1ed9 cao d\u00f9ng \u0111\u1ec3 gi\u1ea3m thi\u1ec3u qu\u00e1 tr\u00ecnh quang h\u00f3a c\u1ee7a fluorophores \u0111\u1ed1i v\u1edbi nh\u1eefng ch\u1ea5t c\u01b0 tr\u00fa trong m\u1eb7t ph\u1eb3ng ti\u00eau c\u1ef1 v\u00e0 do \u0111\u00f3 l\u00e0m gi\u1ea3m \u0111\u1ed9c t\u00ednh quang, l\u00e0m t\u0103ng kh\u1ea3 n\u0103ng t\u1ed3n t\u1ea1i c\u1ee7a m\u1eabu v\u00e0 th\u1eddi gian ti\u1ebfp theo c\u1ee7a c\u00e1c th\u00ed nghi\u1ec7m nghi\u00ean c\u1ee9u t\u00ednh ch\u1ea5t c\u1ee7a t\u1ebf b\u00e0o v\u00e0 m\u00f4 s\u1ed1ng.\u00a0Ngo\u00e0i ra, vi\u1ec7c \u00e1p d\u1ee5ng c\u00e1c b\u01b0\u1edbc s\u00f3ng k\u00edch th\u00edch c\u1eadn h\u1ed3ng ngo\u1ea1i cho ph\u00e9p th\u00e2m nh\u1eadp s\u00e2u h\u01a1n v\u00e0o c\u00e1c v\u1eadt li\u1ec7u sinh h\u1ecdc v\u00e0 gi\u1ea3m m\u1ee9c \u0111\u1ed9 t\u00e1n x\u1ea1 \u00e1nh s\u00e1ng cao \u0111\u01b0\u1ee3c quan s\u00e1t th\u1ea5y \u1edf c\u00e1c b\u01b0\u1edbc s\u00f3ng ng\u1eafn h\u01a1n.<\/p>\n<h2 class=\"paratitle\"><span class=\"ez-toc-section\" id=\"Ung_dung_kinh_hien_vi_huynh_quang\"><\/span><strong>\u1ee8ng d\u1ee5ng <a href=\"https:\/\/vospan.com\/product-brands\/zeiss-vi\/\">k\u00ednh hi\u1ec3n vi hu\u1ef3nh quang\u00a0<\/a><\/strong><span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p><b>K\u00ednh hi\u1ec3n vi hu\u1ef3nh quang ph\u1ea3n x\u1ea1 n\u1ed9i to\u00e0n ph\u1ea7n<\/b> (<b>TIRFM<\/b>) l\u00e0 m\u1ed9t k\u1ef9 thu\u1eadt \u0111\u01b0\u1ee3c thi\u1ebft k\u1ebf \u0111\u1ec3 th\u0103m d\u00f2 b\u1ec1 m\u1eb7t c\u1ee7a c\u00e1c t\u1ebf b\u00e0o s\u1ed1ng \u0111\u01b0\u1ee3c \u0111\u00e1nh d\u1ea5u hu\u1ef3nh quang b\u1eb1ng s\u00f3ng t\u1eaft d\u1ea7n \u0111\u01b0\u1ee3c t\u1ea1o ra b\u1edfi m\u1ed9t ch\u00f9m \u00e1nh s\u00e1ng truy\u1ec1n gi\u1eefa hai m\u00f4i tr\u01b0\u1eddng c\u00f3 chi\u1ebft su\u1ea5t kh\u00e1c nhau. Trong th\u1ef1c t\u1ebf, m\u1ed9t ch\u00f9m tia laze t\u1edbi b\u1ecb ph\u1ea3n x\u1ea1 \u1edf m\u1ed9t g\u00f3c t\u1edbi h\u1ea1n (ph\u1ea3n x\u1ea1 to\u00e0n ph\u1ea7n b\u00ean trong) khi n\u00f3 g\u1eb7p m\u1eb7t ph\u00e2n c\u00e1ch gi\u1eefa l\u1edbp ph\u1ee7 k\u00ednh hi\u1ec3n vi v\u00e0 m\u00f4i tr\u01b0\u1eddng n\u01b0\u1edbc ch\u1ee9a c\u00e1c t\u1ebf b\u00e0o. C\u00e1c hu\u1ef3nh quang trong ph\u1ea1m vi kho\u1ea3ng 10 gi\u00e2y c\u1ee7a b\u1ec1 m\u1eb7t nanomet (t\u1eeb 10 \u0111\u1ebfn 200 nanomet) b\u1ecb k\u00edch th\u00edch b\u1edfi s\u00f3ng bi\u1ebfn thi\u00ean, trong khi nh\u1eefng t\u1ebf b\u00e0o \u1edf xa h\u01a1n h\u1ea7u nh\u01b0 kh\u00f4ng b\u1ecb \u1ea3nh h\u01b0\u1edfng, v\u00ec n\u0103ng l\u01b0\u1ee3ng c\u1ee7a s\u00f3ng bi\u1ebfn thi\u00ean gi\u1ea3m theo c\u1ea5p s\u1ed1 nh\u00e2n v\u1edbi kho\u1ea3ng c\u00e1ch t\u1eeb l\u1edbp ph\u1ee7. Do \u0111\u00f3, TIRFM d\u1eabn \u0111\u1ebfn m\u1ee9c t\u00edn hi\u1ec7u cao ph\u00e1t sinh t\u1eeb c\u00e1c ch\u1ea5t hu\u1ef3nh quang c\u01b0 tr\u00fa g\u1ea7n l\u1edbp ph\u1ee7, \u0111\u01b0\u1ee3c \u0111\u1eb7t ch\u1ed3ng l\u00ean tr\u00ean m\u1ed9t n\u1ec1n r\u1ea5t t\u1ed1i, cung c\u1ea5p t\u1ef7 l\u1ec7 t\u00edn hi\u1ec7u tr\u00ean t\u1ea1p \u00e2m t\u1ed1t nh\u1ea5t c\u00f3 th\u1ec3. Gi\u1edbi h\u1ea1n c\u1ef1c \u0111\u1ed9 v\u1ec1 \u0111\u1ed9 s\u00e2u k\u00edch th\u00edch l\u00e0 l\u00fd t\u01b0\u1edfng \u0111\u1ec3 nghi\u00ean c\u1ee9u c\u00e1c ph\u00e2n t\u1eed \u0111\u01a1n l\u1ebb ho\u1eb7c c\u00e1c th\u00e0nh ph\u1ea7n m\u00e0ng v\u00e0 c\u01a1 quan g\u1ea7n b\u1ec1 m\u1eb7t c\u1ee7a l\u1edbp ph\u1ee7 trong c\u00e1c t\u1ebf b\u00e0o k\u1ebft d\u00ednh (xem H\u00ecnh 8(e)). V\u00ec s\u1ef1 k\u00edch th\u00edch \u0111\u01b0\u1ee3c gi\u1edbi h\u1ea1n \u1edf v\u00f9ng m\u1ecfng ti\u1ebfp gi\u00e1p v\u1edbi l\u1edbp ph\u1ee7, qu\u00e1 tr\u00ecnh t\u1ea9y tr\u1eafng v\u00e0 \u0111\u1ed9c t\u00ednh quang c\u0169ng b\u1ecb gi\u1edbi h\u1ea1n \u1edf nh\u1eefng khu v\u1ef1c n\u00e0y, khi\u1ebfn TIRFM tr\u1edf th\u00e0nh m\u1ed9t trong nh\u1eefng ph\u01b0\u01a1ng ph\u00e1p h\u1eefu \u00edch nh\u1ea5t cho c\u00e1c quan s\u00e1t d\u00e0i h\u1ea1n. K\u1ef9 thu\u1eadt n\u00e0y \u0111\u00e3 tr\u1edf th\u00e0nh m\u1ed9t c\u00f4ng c\u1ee5 c\u01a1 b\u1ea3n \u0111\u1ec3 \u0111i\u1ec1u tra m\u1ed9t lo\u1ea1t c\u00e1c hi\u1ec7n t\u01b0\u1ee3ng trong t\u1ebf b\u00e0o v\u00e0 sinh h\u1ecdc ph\u00e2n t\u1eed. V\u00ec s\u1ef1 k\u00edch th\u00edch \u0111\u01b0\u1ee3c gi\u1edbi h\u1ea1n \u1edf v\u00f9ng m\u1ecfng ti\u1ebfp gi\u00e1p v\u1edbi l\u1edbp ph\u1ee7, qu\u00e1 tr\u00ecnh t\u1ea9y tr\u1eafng v\u00e0 \u0111\u1ed9c t\u00ednh quang c\u0169ng b\u1ecb gi\u1edbi h\u1ea1n \u1edf nh\u1eefng khu v\u1ef1c n\u00e0y, khi\u1ebfn TIRFM tr\u1edf th\u00e0nh m\u1ed9t trong nh\u1eefng ph\u01b0\u01a1ng ph\u00e1p h\u1eefu \u00edch nh\u1ea5t cho c\u00e1c quan s\u00e1t d\u00e0i h\u1ea1n. K\u1ef9 thu\u1eadt n\u00e0y \u0111\u00e3 tr\u1edf th\u00e0nh m\u1ed9t c\u00f4ng c\u1ee5 c\u01a1 b\u1ea3n \u0111\u1ec3 \u0111i\u1ec1u tra m\u1ed9t lo\u1ea1t c\u00e1c hi\u1ec7n t\u01b0\u1ee3ng trong t\u1ebf b\u00e0o v\u00e0 sinh h\u1ecdc ph\u00e2n t\u1eed. V\u00ec s\u1ef1 k\u00edch th\u00edch \u0111\u01b0\u1ee3c gi\u1edbi h\u1ea1n \u1edf v\u00f9ng m\u1ecfng ti\u1ebfp gi\u00e1p v\u1edbi l\u1edbp ph\u1ee7, qu\u00e1 tr\u00ecnh t\u1ea9y tr\u1eafng v\u00e0 \u0111\u1ed9c t\u00ednh quang c\u0169ng b\u1ecb gi\u1edbi h\u1ea1n \u1edf nh\u1eefng khu v\u1ef1c n\u00e0y, khi\u1ebfn TIRFM tr\u1edf th\u00e0nh m\u1ed9t trong nh\u1eefng ph\u01b0\u01a1ng ph\u00e1p h\u1eefu \u00edch nh\u1ea5t cho c\u00e1c quan s\u00e1t d\u00e0i h\u1ea1n. K\u1ef9 thu\u1eadt n\u00e0y \u0111\u00e3 tr\u1edf th\u00e0nh m\u1ed9t c\u00f4ng c\u1ee5 c\u01a1 b\u1ea3n \u0111\u1ec3 \u0111i\u1ec1u tra m\u1ed9t lo\u1ea1t c\u00e1c hi\u1ec7n t\u01b0\u1ee3ng trong t\u1ebf b\u00e0o v\u00e0 sinh h\u1ecdc ph\u00e2n t\u1eed.<\/p>\n<p><b>Ph\u00e2n t\u00edch gi\u1ea3i ch\u1eadp<\/b>\u00a0l\u00e0 m\u1ed9t k\u1ef9 thu\u1eadt \u00e1p d\u1ee5ng c\u00e1c thu\u1eadt to\u00e1n cho m\u1ed9t ch\u1ed3ng h\u00ecnh \u1ea3nh l\u1ea5y n\u00e9t xuy\u00ean su\u1ed1t thu \u0111\u01b0\u1ee3c d\u1ecdc theo quang h\u1ecdc (\u00a0<b>z<\/b>) \u0111\u1ec3 t\u0103ng c\u01b0\u1eddng t\u00edn hi\u1ec7u photon c\u1ee5 th\u1ec3 cho m\u1ed9t m\u1eb7t ph\u1eb3ng h\u00ecnh \u1ea3nh nh\u1ea5t \u0111\u1ecbnh ho\u1eb7c nhi\u1ec1u m\u1eb7t ph\u1eb3ng ti\u00eau c\u1ef1 trong ch\u1ed3ng h\u00ecnh \u1ea3nh.\u00a0K\u00ednh hi\u1ec3n vi ph\u1ea3i \u0111\u01b0\u1ee3c trang b\u1ecb m\u1ed9t \u1ed5 l\u1ea5y n\u00e9t c\u01a1 gi\u1edbi c\u00f3 \u0111\u1ed9 ch\u00ednh x\u00e1c cao \u0111\u1ec3 \u0111\u1ea3m b\u1ea3o thu \u0111\u01b0\u1ee3c h\u00ecnh \u1ea3nh \u1edf c\u00e1c kho\u1ea3ng \u0111\u01b0\u1ee3c x\u00e1c \u0111\u1ecbnh ch\u00ednh x\u00e1c gi\u1eefa c\u00e1c m\u1eb7t ph\u1eb3ng ti\u00eau c\u1ef1 trong m\u1eabu v\u1eadt.\u00a0Trong m\u1ed9t \u1ee9ng d\u1ee5ng \u0111i\u1ec3n h\u00ecnh (xem H\u00ecnh 8(f)), ph\u00e2n t\u00edch gi\u1ea3i ch\u1eadp \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng \u0111\u1ec3 l\u00e0m m\u1edd v\u00e0 lo\u1ea1i b\u1ecf \u00e1nh s\u00e1ng n\u1eb1m ngo\u00e0i v\u00f9ng l\u1ea5y n\u00e9t kh\u1ecfi m\u1ed9t m\u1eb7t ph\u1eb3ng ti\u00eau \u0111i\u1ec3m quan t\u00e2m c\u1ee5 th\u1ec3 b\u1eb1ng c\u00e1ch s\u1eed d\u1ee5ng k\u00edch th\u00edch v\u00e0 ph\u00e1t x\u1ea1 hu\u1ef3nh quang tr\u01b0\u1eddng r\u1ed9ng (m\u1eb7c d\u00f9 k\u1ef9 thu\u1eadt n\u00e0y h\u1eefu \u00edch cho c\u00e1c ch\u1ebf \u0111\u1ed9 chi\u1ebfu s\u00e1ng kh\u00e1c c\u0169ng).\u00a0C\u00e1c \u1ee9ng d\u1ee5ng ph\u1ee9c t\u1ea1p nh\u1ea5t \u00e1p d\u1ee5ng ph\u00e2n t\u00edch gi\u1ea3i ch\u1eadp cho to\u00e0n b\u1ed9 ng\u0103n x\u1ebfp h\u00ecnh \u1ea3nh \u0111\u1ec3 t\u1ea1o ra c\u00e1c h\u00ecnh chi\u1ebfu ho\u1eb7c m\u00f4 h\u00ecnh ba chi\u1ec1u.\u00a0T\u1eadp h\u1ee3p c\u00e1c h\u00ecnh \u1ea3nh tr\u01b0\u1eddng r\u1ed9ng \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng \u0111\u1ec3 ph\u00e2n t\u00edch gi\u1ea3i ch\u1eadp s\u1ebd ghi l\u1ea1i s\u1ed1 l\u01b0\u1ee3ng photon t\u1ed1i \u0111a theo l\u00fd thuy\u1ebft m\u00e0 m\u1eabu v\u1eadt ph\u00e1t ra.\u00a0Qu\u00e1 tr\u00ecnh gi\u1ea3i ch\u1eadp ch\u1ec9 \u0111\u1ecbnh l\u1ea1i c\u01b0\u1eddng \u0111\u1ed9 &#8220;m\u1edd&#8221; ph\u00e1t sinh t\u1eeb c\u00e1c photon ph\u00e1t ra b\u00ean tr\u00ean v\u00e0 b\u00ean d\u01b0\u1edbi m\u1eb7t ph\u1eb3ng ti\u00eau \u0111i\u1ec3m cho m\u1eb7t ph\u1eb3ng g\u1ed1c.\u00a0Do \u0111\u00f3, gi\u1ea3i ch\u1eadp s\u1eed d\u1ee5ng th\u1ef1c t\u1ebf t\u1ea5t c\u1ea3 c\u01b0\u1eddng \u0111\u1ed9 ph\u00e1t x\u1ea1 c\u00f3 s\u1eb5n v\u00e0 cung c\u1ea5p ng\u00e2n s\u00e1ch \u00e1nh s\u00e1ng t\u1ed1t nh\u1ea5t c\u00f3 th\u1ec3, do \u0111\u00f3 l\u00e0m cho k\u1ef9 thu\u1eadt n\u00e0y tr\u1edf th\u00e0nh ph\u01b0\u01a1ng ph\u00e1p \u0111\u01b0\u1ee3c l\u1ef1a ch\u1ecdn cho c\u00e1c m\u1eabu c\u1ef1c k\u1ef3 nh\u1ea1y s\u00e1ng.<\/p>\n<p><img loading=\"lazy\" decoding=\"async\" class=\"aligncenter\" src=\"https:\/\/zeiss-campus.magnet.fsu.edu\/articles\/livecellimaging\/images\/techniquesfigure8.jpg\" width=\"439\" height=\"340\" border=\"0\" \/>S\u1ef1 th\u00edch \u1ee9ng c\u1ee7a hi\u1ec7n t\u01b0\u1ee3ng truy\u1ec1n n\u0103ng l\u01b0\u1ee3ng c\u1ed9ng h\u01b0\u1edfng sang k\u00ednh hi\u1ec3n vi hu\u1ef3nh quang,\u00a0<b>hu\u1ef3nh quang ho\u1eb7c truy\u1ec1n n\u0103ng l\u01b0\u1ee3ng c\u1ed9ng h\u01b0\u1edfng Forster<\/b> (<b>FRET<\/b>) \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng \u0111\u1ec3 thu \u0111\u01b0\u1ee3c th\u00f4ng tin \u0111\u1ecbnh l\u01b0\u1ee3ng v\u1ec1 th\u1eddi gian v\u00e0 kh\u00f4ng gian v\u1ec1 s\u1ef1 li\u00ean k\u1ebft v\u00e0 t\u01b0\u01a1ng t\u00e1c c\u1ee7a protein, lipid, enzyme v\u00e0 axit nucleic trong t\u1ebf b\u00e0o s\u1ed1ng.\u00a0K\u00ednh hi\u1ec3n vi FRET \u0111\u01b0\u1ee3c th\u1ef1c hi\u1ec7n b\u1eb1ng c\u00e1ch s\u1eed d\u1ee5ng tr\u1ea1ng th\u00e1i \u1ed5n \u0111\u1ecbnh ho\u1eb7c k\u1ef9 thu\u1eadt gi\u1ea3i quy\u1ebft th\u1eddi gian, nh\u01b0ng h\u00ecnh \u1ea3nh FRET \u0111\u01b0\u1ee3c gi\u1ea3i quy\u1ebft theo th\u1eddi gian c\u00f3 l\u1ee3i th\u1ebf l\u00e0 \u00e1nh x\u1ea1 ch\u00ednh x\u00e1c h\u01a1n kho\u1ea3ng c\u00e1ch ng\u01b0\u1eddi cho-ng\u01b0\u1eddi nh\u1eadn.\u00a0M\u1ed9t k\u00ednh hi\u1ec3n vi hu\u1ef3nh quang tr\u01b0\u1eddng r\u1ed9ng ti\u00eau chu\u1ea9n \u0111\u01b0\u1ee3c trang b\u1ecb c\u00e1c b\u1ed9 l\u1ecdc k\u00edch th\u00edch v\u00e0 ph\u00e1t x\u1ea1 th\u00edch h\u1ee3p v\u00e0 m\u1ed9t m\u00e1y quay video nh\u1ea1y c\u1ea3m c\u00f3 th\u1ec3 \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng \u0111\u1ec3 th\u1ef1c hi\u1ec7n h\u00ecnh \u1ea3nh FRET.\u00a0C\u00e1c c\u1ea3m bi\u1ebfn sinh h\u1ecdc k\u1eb9p m\u1ed9t protein ho\u1eb7c peptide nh\u1ea1y c\u1ea3m v\u1edbi m\u00f4i tr\u01b0\u1eddng gi\u1eefa hai protein ph\u00e1t hu\u1ef3nh quang c\u00f3 kh\u1ea3 n\u0103ng FRET hi\u1ec7n \u0111ang \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng r\u1ed9ng r\u00e3i trong sinh h\u1ecdc t\u1ebf b\u00e0o.\u00a0Nh\u1eefng \u0111\u1ea7u d\u00f2 n\u00e0y d\u1ec5 d\u00e0ng \u0111\u01b0\u1ee3c ch\u1ee5p \u1ea3nh trong k\u00ednh hi\u1ec3n vi hu\u1ef3nh quang tr\u01b0\u1eddng r\u1ed9ng b\u1eb1ng c\u00e1ch s\u1eed d\u1ee5ng c\u00e1c k\u1ef9 thu\u1eadt FRET ph\u00e1t x\u1ea1 nh\u1ea1y c\u1ea3m k\u1ebft h\u1ee3p v\u1edbi ph\u00e2n t\u00edch \u0111o l\u01b0\u1eddng.<\/p>\n<p><b>K\u00ednh hi\u1ec3n vi h\u00ecnh \u1ea3nh th\u1eddi gian t\u1ed3n t\u1ea1i hu\u1ef3nh quang <\/b>(<b>FLIM<\/b>) l\u00e0 m\u1ed9t k\u1ef9 thu\u1eadt ph\u1ee9c t\u1ea1p cho ph\u00e9p ghi l\u1ea1i \u0111\u1ed3ng th\u1eddi c\u1ea3 th\u1eddi gian t\u1ed3n t\u1ea1i c\u1ee7a hu\u1ef3nh quang v\u00e0 v\u1ecb tr\u00ed kh\u00f4ng gian c\u1ee7a c\u00e1c ch\u1ea5t hu\u1ef3nh quang tr\u00ean m\u1ecdi v\u1ecb tr\u00ed trong \u1ea3nh.\u00a0Ph\u01b0\u01a1ng ph\u00e1p n\u00e0y cung c\u1ea5p m\u1ed9t c\u01a1 ch\u1ebf \u0111\u1ec3 nghi\u00ean c\u1ee9u c\u00e1c th\u00f4ng s\u1ed1 m\u00f4i tr\u01b0\u1eddng nh\u01b0 pH, n\u1ed3ng \u0111\u1ed9 ion, \u0111\u1ed9 ph\u00e2n c\u1ef1c c\u1ee7a dung m\u00f4i, t\u01b0\u01a1ng t\u00e1c kh\u00f4ng c\u1ed9ng h\u00f3a tr\u1ecb, \u0111\u1ed9 nh\u1edbt v\u00e0 s\u1ee9c c\u0103ng oxy trong c\u00e1c t\u1ebf b\u00e0o s\u1ed1ng \u0111\u01a1n l\u1ebb, tr\u00ecnh b\u00e0y d\u1eef li\u1ec7u theo m\u1ea3ng kh\u00f4ng gian v\u00e0 th\u1eddi gian.\u00a0C\u00e1c ph\u00e9p \u0111o FLIM v\u1ec1 th\u1eddi gian t\u1ed3n t\u1ea1i \u1edf tr\u1ea1ng th\u00e1i k\u00edch th\u00edch nano gi\u00e2y kh\u00f4ng ph\u1ee5 thu\u1ed9c v\u00e0o n\u1ed3ng \u0111\u1ed9 fluorophore c\u1ee5c b\u1ed9, t\u1ea1o t\u00e1c t\u1ea9y tr\u1eafng b\u1eb1ng quang h\u00f3a v\u00e0 \u0111\u1ed9 d\u00e0i \u0111\u01b0\u1eddng d\u1eabn (\u0111\u1ed9 d\u00e0y m\u1eabu v\u1eadt), nh\u01b0ng nh\u1ea1y c\u1ea3m v\u1edbi c\u00e1c ph\u1ea3n \u1ee9ng \u1edf tr\u1ea1ng th\u00e1i k\u00edch th\u00edch nh\u01b0 truy\u1ec1n n\u0103ng l\u01b0\u1ee3ng c\u1ed9ng h\u01b0\u1edfng.\u00a0Trong th\u1ef1c t\u1ebf,<\/p>\n<p>\u0110\u1ed9 linh \u0111\u1ed9ng t\u1ecbnh ti\u1ebfn (h\u1ec7 s\u1ed1 khu\u1ebfch t\u00e1n b\u00ean) c\u1ee7a c\u00e1c \u0111\u1ea1i ph\u00e2n t\u1eed \u0111\u01b0\u1ee3c \u0111\u00e1nh d\u1ea5u hu\u1ef3nh quang v\u00e0 c\u00e1c ch\u1ea5t ph\u00e1t hu\u1ef3nh quang nh\u1ecf c\u00f3 th\u1ec3 \u0111\u01b0\u1ee3c x\u00e1c \u0111\u1ecbnh b\u1eb1ng\u00a0<b>kh\u1ea3 n\u0103ng ph\u1ee5c h\u1ed3i hu\u1ef3nh quang sau khi t\u1ea9y tr\u1eafng quang h\u1ecdc<\/b>(<b>FRAP<\/b>) k\u1ef9 thu\u1eadt.\u00a0Trong FRAP, m\u1ed9t v\u00f9ng r\u1ea5t nh\u1ecf, \u0111\u01b0\u1ee3c ch\u1ecdn (\u0111\u01b0\u1eddng k\u00ednh v\u00e0i micromet) \u0111\u01b0\u1ee3c chi\u1ebfu s\u00e1ng c\u01b0\u1eddng \u0111\u1ed9 cao, th\u01b0\u1eddng l\u00e0 b\u1eb1ng tia laser, \u0111\u1ec3 t\u1ea1o ra qu\u00e1 tr\u00ecnh quang h\u00f3a ho\u00e0n to\u00e0n c\u00e1c fluorophores trong v\u00f9ng.\u00a0K\u1ebft qu\u1ea3 l\u00e0 l\u00e0m gi\u1ea3m \u0111\u00e1ng k\u1ec3 ho\u1eb7c tri\u1ec7t ti\u00eau hu\u1ef3nh quang.\u00a0Sau xung t\u1ea9y tr\u1eafng b\u1eb1ng quang h\u00f3a, t\u1ed1c \u0111\u1ed9 v\u00e0 m\u1ee9c \u0111\u1ed9 ph\u1ee5c h\u1ed3i c\u01b0\u1eddng \u0111\u1ed9 hu\u1ef3nh quang \u1edf v\u00f9ng \u0111\u01b0\u1ee3c t\u1ea9y tr\u1eafng \u0111\u01b0\u1ee3c theo d\u00f5i nh\u01b0 m\u1ed9t h\u00e0m c\u1ee7a th\u1eddi gian \u1edf c\u01b0\u1eddng \u0111\u1ed9 k\u00edch th\u00edch th\u1ea5p h\u01a1n \u0111\u1ec3 t\u1ea1o ra th\u00f4ng tin v\u1ec1 qu\u00e1 tr\u00ecnh t\u00e1i t\u1ea1o d\u00e2n s\u1ed1 b\u1eb1ng fluorophores v\u00e0 \u0111\u1ed9ng h\u1ecdc c\u1ee7a qu\u00e1 tr\u00ecnh ph\u1ee5c h\u1ed3i (<em>H\u00ecnh 9<\/em>).\u00a0FRAP th\u01b0\u1eddng \u0111\u01b0\u1ee3c ti\u1ebfn h\u00e0nh b\u1eb1ng c\u00e1ch s\u1eed d\u1ee5ng EGFP ho\u1eb7c c\u00e1c protein hu\u1ef3nh quang kh\u00e1c.\u00a0C\u00e1c k\u1ef9 thu\u1eadt k\u00edch ho\u1ea1t quang li\u00ean quan d\u1ef1a tr\u00ean c\u00e1c ch\u1ea5t hu\u1ef3nh quang t\u1ed5ng h\u1ee3p \u0111\u01b0\u1ee3c l\u1ed3ng trong l\u1ed3ng chuy\u00ean d\u1ee5ng ho\u1eb7c c\u00e1c protein hu\u1ef3nh quang \u0111\u01b0\u1ee3c \u01b0u \u0111\u00e3i t\u01b0\u01a1ng t\u1ef1 c\u00f3 th\u1ec3 \u0111\u01b0\u1ee3c k\u00edch ho\u1ea1t b\u1eb1ng m\u1ed9t xung ng\u1eafn c\u1ee7a tia c\u1ef1c t\u00edm ho\u1eb7c \u00e1nh s\u00e1ng t\u00edm.<\/p>\n<p><img loading=\"lazy\" decoding=\"async\" class=\"aligncenter\" src=\"https:\/\/zeiss-campus.magnet.fsu.edu\/articles\/livecellimaging\/images\/techniquesfigure9.jpg\" width=\"566\" height=\"283\" border=\"0\" \/>Trong m\u1ed9t k\u1ef9 thu\u1eadt li\u00ean quan \u0111\u1ebfn FRAP (<b>kh\u00f4ng th\u1ec3 ph\u00e1t quang trong qu\u00e1 tr\u00ecnh t\u1ea9y tr\u1eafng b\u1eb1ng \u00e1nh s\u00e1ng <\/b>; <b>FLIP<\/b>\u00a0), m\u1ed9t v\u00f9ng hu\u1ef3nh quang x\u00e1c \u0111\u1ecbnh trong m\u1ed9t t\u1ebf b\u00e0o s\u1ed1ng ph\u1ea3i ch\u1ecbu qu\u00e1 tr\u00ecnh t\u1ea9y tr\u1eafng l\u1eb7p \u0111i l\u1eb7p l\u1ea1i b\u1eb1ng c\u00e1ch chi\u1ebfu s\u00e1ng b\u1eb1ng b\u1ee9c x\u1ea1 c\u01b0\u1eddng \u0111\u1ed9 cao.\u00a0Trong m\u1ed9t kho\u1ea3ng th\u1eddi gian \u0111o \u0111\u01b0\u1ee3c, h\u00e0nh \u0111\u1ed9ng n\u00e0y s\u1ebd d\u1eabn \u0111\u1ebfn m\u1ea5t ho\u00e0n to\u00e0n t\u00edn hi\u1ec7u hu\u1ef3nh quang trong to\u00e0n b\u1ed9 t\u1ebf b\u00e0o n\u1ebfu t\u1ea5t c\u1ea3 c\u00e1c ch\u1ea5t ph\u00e1t hu\u1ef3nh quang c\u00f3 th\u1ec3 khu\u1ebfch t\u00e1n v\u00e0o v\u00f9ng \u0111ang \u0111\u01b0\u1ee3c t\u1ea9y tr\u1eafng quang h\u00f3a.\u00a0B\u1eb1ng c\u00e1ch t\u00ednh to\u00e1n t\u1ed1c \u0111\u1ed9 m\u00e0 hu\u1ef3nh quang b\u1ecb d\u1eadp t\u1eaft kh\u1ecfi to\u00e0n b\u1ed9 t\u1ebf b\u00e0o, c\u00f3 th\u1ec3 x\u00e1c \u0111\u1ecbnh \u0111\u01b0\u1ee3c \u0111\u1ed9 linh \u0111\u1ed9ng khu\u1ebfch t\u00e1n c\u1ee7a ch\u1ea5t hu\u1ef3nh quang m\u1ee5c ti\u00eau.\u00a0H\u01a1n n\u1eefa, FLIP s\u1ebd d\u1ec5 d\u00e0ng x\u00e1c \u0111\u1ecbnh v\u1ecb tr\u00ed v\u00e0 b\u1ea3n ch\u1ea5t c\u1ee7a b\u1ea5t k\u1ef3 r\u00e0o c\u1ea3n khu\u1ebfch t\u00e1n n\u00e0o gi\u1eefa c\u00e1c ng\u0103n ri\u00eang l\u1ebb c\u1ee7a t\u1ebf b\u00e0o, ch\u1eb3ng h\u1ea1n nh\u01b0 r\u00e0o c\u1ea3n gi\u1eefa soma v\u00e0 s\u1ee3i tr\u1ee5c c\u1ee7a n\u01a1-ron.<\/p>\n<p>Vi\u1ec7c s\u1eed d\u1ee5ng ng\u00e0y c\u00e0ng nhi\u1ec1u protein hu\u1ef3nh quang v\u1edbi ph\u1ed5 ph\u00e1t x\u1ea1 ch\u1ed3ng ch\u00e9o cao trong h\u00ecnh \u1ea3nh t\u1ebf b\u00e0o s\u1ed1ng th\u01b0\u1eddng \u0111\u00f2i h\u1ecfi c\u00e1c gi\u1ea3i ph\u00e1p \u0111\u1ed9c \u0111\u00e1o \u0111\u1ec3 cung c\u1ea5p s\u1ef1 ph\u00e2n t\u00e1ch \u0111\u1ea7y \u0111\u1ee7 c\u00e1c t\u00edn hi\u1ec7u t\u1eeb c\u00e1c m\u1ee5c ti\u00eau kh\u00e1c nhau.\u00a0<b>H\u00ecnh \u1ea3nh quang ph\u1ed5<\/b>\u00a0d\u1ef1a tr\u00ean ph\u1ea7n c\u1ee9ng chuy\u00ean d\u1ee5ng (ch\u1ee7 y\u1ebfu \u0111\u01b0\u1ee3c k\u1ebft h\u1ee3p v\u1edbi k\u00ednh hi\u1ec3n vi \u0111\u1ed3ng ti\u00eau) \u0111\u1ec3 t\u00e1ch \u00e1nh s\u00e1ng ph\u00e1t x\u1ea1 th\u00e0nh c\u00e1c th\u00e0nh ph\u1ea7n quang ph\u1ed5 c\u1ee7a n\u00f3.\u00a0<b>Unmixing tuy\u1ebfn t\u00ednh <\/b>l\u00e0 m\u1ed9t quy tr\u00ecnh t\u00ednh to\u00e1n li\u00ean quan \u0111\u1ebfn gi\u1ea3i ch\u1eadp s\u1eed d\u1ee5ng c\u1ea5u h\u00ecnh quang ph\u1ed5 duy nh\u1ea5t c\u1ee7a t\u1eebng ch\u1ea5t hu\u1ef3nh quang trong m\u1eabu v\u1eadt \u0111\u1ec3 g\u00e1n l\u1ea1i t\u00edn hi\u1ec7u cho c\u00e1c pixel th\u00edch h\u1ee3p trong h\u00ecnh \u1ea3nh cu\u1ed1i c\u00f9ng.\u00a0M\u1eb7c d\u00f9 c\u00e1c c\u00f4ng c\u1ee5 ph\u00e2n t\u00edch n\u00e0y c\u00f3 th\u1ec3 \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng c\u00f9ng nhau \u0111\u1ec3 ph\u00e2n bi\u1ec7t gi\u1eefa c\u00e1c ch\u1ea5t ph\u00e1t quang ri\u00eang bi\u1ec7t c\u00f3 ph\u1ed5 ch\u1ed3ng l\u1ea5p cao, nh\u01b0ng ch\u00fang ho\u1ea1t \u0111\u1ed9ng v\u1edbi c\u00e1i gi\u00e1 ph\u1ea3i tr\u1ea3 l\u00e0 y\u00eau c\u1ea7u ph\u00e1t hi\u1ec7n nhi\u1ec1u photon h\u01a1n \u0111\u00e1ng k\u1ec3 t\u1eeb m\u1ed7i pixel, \u0111i\u1ec1u n\u00e0y c\u00f3 th\u1ec3 g\u00e2y kh\u00f3 kh\u0103n cho vi\u1ec7c ch\u1ee5p \u1ea3nh t\u1ebf b\u00e0o s\u1ed1ng c\u1ee7a m\u1eabu v\u1eadt \u0111\u01b0\u1ee3c d\u00e1n nh\u00e3n ch\u1ea5t ph\u00e1t quang m\u1edd ho\u1eb7c c\u00e1c m\u1ee5c ti\u00eau phong ph\u00fa th\u1ea5p.<\/p>\n<p>\u0110\u01b0\u1ee3c s\u1eed d\u1ee5ng ch\u1ee7 y\u1ebfu v\u1edbi k\u00ednh hi\u1ec3n vi qu\u00e9t \u0111\u1ed3ng ti\u00eau ho\u1eb7c \u0111a \u0111i\u1ec3m qu\u00e9t laser, <b>ph\u1ed5 t\u01b0\u01a1ng quan hu\u1ef3nh quang<\/b> (<b>FCS<\/b>) l\u00e0 m\u1ed9t k\u1ef9 thu\u1eadt \u0111\u01b0\u1ee3c thi\u1ebft k\u1ebf \u0111\u1ec3 x\u00e1c \u0111\u1ecbnh \u0111\u1ed9ng l\u1ef1c h\u1ecdc ph\u00e2n t\u1eed trong c\u00e1c th\u1ec3 t\u00edch ch\u1ec9 ch\u1ee9a m\u1ed9t ho\u1eb7c m\u1ed9t v\u00e0i ph\u00e2n t\u1eed, mang l\u1ea1i th\u00f4ng tin v\u1ec1 t\u1ed1c \u0111\u1ed9 ph\u1ea3n \u1ee9ng h\u00f3a h\u1ecdc, h\u1ec7 s\u1ed1 khu\u1ebfch t\u00e1n, tr\u1ecdng l\u01b0\u1ee3ng ph\u00e2n t\u1eed, t\u1ed1c \u0111\u1ed9 d\u00f2ng ch\u1ea3y v\u00e0 s\u1ef1 k\u1ebft t\u1ee5.\u00a0Trong FCS, m\u1ed9t th\u1ec3 t\u00edch nh\u1ecf (x\u1ea5p x\u1ec9 m\u1ed9t femtolit; ti\u00eau \u0111i\u1ec3m gi\u1edbi h\u1ea1n nhi\u1ec5u x\u1ea1 c\u1ee7a ch\u00f9m tia laze) \u0111\u01b0\u1ee3c chi\u1ebfu x\u1ea1 b\u1eb1ng ch\u00f9m tia laze h\u1ed9i t\u1ee5 \u0111\u1ec3 ghi l\u1ea1i c\u00e1c dao \u0111\u1ed9ng c\u01b0\u1eddng \u0111\u1ed9 hu\u1ef3nh quang t\u1ef1 ph\u00e1t ph\u00e1t sinh t\u1eeb \u0111\u1ed9ng l\u1ef1c h\u1ecdc c\u1ee7a c\u00e1c ph\u00e2n t\u1eed hu\u1ef3nh quang chi\u1ebfm th\u1ec3 t\u00edch nh\u01b0 m\u1ed9t h\u00e0m c\u1ee7a th\u1eddi gian ( xem H\u00ecnh 10).\u00a0C\u00e1c hu\u1ef3nh quang t\u01b0\u01a1ng \u0111\u1ed1i nh\u1ecf khu\u1ebfch t\u00e1n nhanh ch\u00f3ng qua th\u1ec3 t\u00edch \u0111\u01b0\u1ee3c chi\u1ebfu s\u00e1ng \u0111\u1ec3 t\u1ea1o ra c\u00e1c \u0111\u1ee3t c\u01b0\u1eddng \u0111\u1ed9 ng\u1eafn, ng\u1eabu nhi\u00ean.\u00a0Ng\u01b0\u1ee3c l\u1ea1i, c\u00e1c ph\u1ee9c h\u1ee3p l\u1edbn h\u01a1n (c\u00e1c ch\u1ea5t hu\u1ef3nh quang li\u00ean k\u1ebft v\u1edbi c\u00e1c \u0111\u1ea1i ph\u00e2n t\u1eed) di chuy\u1ec3n ch\u1eadm h\u01a1n v\u00e0 t\u1ea1o ra ki\u1ec3u c\u01b0\u1eddng \u0111\u1ed9 hu\u1ef3nh quang ph\u1ee5 thu\u1ed9c v\u00e0o th\u1eddi gian l\u00e2u h\u01a1n, b\u1ec1n v\u1eefng h\u01a1n.<\/p>\n<p>\u0110\u1ed9ng l\u1ef1c v\u1ed1n c\u00f3 v\u00e0 s\u1ef1 ph\u00e2n b\u1ed1 kh\u00f4ng gian c\u1ee7a c\u00e1c c\u1ea5u tr\u00fac \u0111\u01b0\u1ee3c d\u00e1n nh\u00e3n hu\u1ef3nh quang c\u00f3 th\u1ec3 kh\u00f3 ph\u00e2n t\u00edch khi c\u00e1c th\u1ef1c th\u1ec3 n\u00e0y \u0111\u01b0\u1ee3c \u0111\u00f3ng g\u00f3i d\u00e0y \u0111\u1eb7c v\u00e0 ch\u1ed3ng ch\u00e9o trong c\u00e1c v\u00f9ng c\u1ee5 th\u1ec3 c\u1ee7a t\u1ebf b\u00e0o s\u1ed1ng.\u00a0<b>K\u00ednh hi\u1ec3n vi \u0111\u1ed1m hu\u1ef3nh quang<\/b> (<b>FSM<\/b>) l\u00e0 m\u1ed9t k\u1ef9 thu\u1eadt t\u01b0\u01a1ng th\u00edch v\u1edbi h\u1ea7u h\u1ebft t\u1ea5t c\u1ea3 c\u00e1c ph\u01b0\u01a1ng th\u1ee9c h\u00ecnh \u1ea3nh t\u1eadn d\u1ee5ng n\u1ed3ng \u0111\u1ed9 r\u1ea5t th\u1ea5p c\u1ee7a c\u00e1c ti\u1ec3u \u0111\u01a1n v\u1ecb \u0111\u01b0\u1ee3c \u0111\u00e1nh d\u1ea5u hu\u1ef3nh quang \u0111\u1ec3 gi\u1ea3m hi\u1ec7n t\u01b0\u1ee3ng hu\u1ef3nh quang ngo\u00e0i ti\u00eau \u0111i\u1ec3m v\u00e0 c\u1ea3i thi\u1ec7n kh\u1ea3 n\u0103ng hi\u1ec3n th\u1ecb c\u1ee7a c\u00e1c c\u1ea5u tr\u00fac \u0111\u01b0\u1ee3c \u0111\u00e1nh d\u1ea5u v\u00e0 \u0111\u1ed9ng l\u1ef1c h\u1ecdc c\u1ee7a ch\u00fang trong c\u00e1c v\u00f9ng d\u00e0y.\u00a0FSM \u0111\u01b0\u1ee3c th\u1ef1c hi\u1ec7n b\u1eb1ng c\u00e1ch ch\u1ec9 d\u00e1n nh\u00e3n m\u1ed9t ph\u1ea7n c\u1ee7a to\u00e0n b\u1ed9 c\u1ea5u tr\u00fac quan t\u00e2m.\u00a0Theo ngh\u0129a \u0111\u00f3, n\u00f3 t\u01b0\u01a1ng t\u1ef1 nh\u01b0 vi\u1ec7c th\u1ef1c hi\u1ec7n FCS tr\u00ean to\u00e0n b\u1ed9 tr\u01b0\u1eddng nh\u00ecn, m\u1eb7c d\u00f9 nh\u1ea5n m\u1ea1nh h\u01a1n v\u00e0o c\u00e1c m\u1eabu kh\u00f4ng gian h\u01a1n l\u00e0 ph\u00e2n t\u00edch th\u1eddi gian \u0111\u1ecbnh l\u01b0\u1ee3ng.\u00a0K\u00ednh hi\u1ec3n vi \u0111\u1ed1m (nh\u01b0 n\u00f3 th\u01b0\u1eddng \u0111\u01b0\u1ee3c g\u1ecdi) \u0111\u1eb7c bi\u1ec7t h\u1eefu \u00edch trong vi\u1ec7c x\u00e1c \u0111\u1ecbnh t\u00ednh di \u0111\u1ed9ng v\u00e0 tr\u00f9ng h\u1ee3p c\u1ee7a c\u00e1c y\u1ebfu t\u1ed1 khung t\u1ebf b\u00e0o, ch\u1eb3ng h\u1ea1n nh\u01b0 actin v\u00e0 vi \u1ed1ng, trong c\u00e1c t\u1ebf b\u00e0o c\u00f3 t\u00ednh di \u0111\u1ed9ng cao.<\/p>\n<p>L\u1ee3i \u00edch ch\u00ednh c\u1ee7a k\u00ednh\u00a0hi\u1ec3n vi <b>t\u00e1n x\u1ea1\u00a0Raman ch\u1ed1ng Stokes k\u1ebft h\u1ee3p<\/b>\u00a0(<b>CARS<\/b>) l\u00e0 n\u00f3 cho ph\u00e9p nghi\u00ean c\u1ee9u c\u00e1c ph\u00e2n t\u1eed sinh h\u1ecdc m\u00e0 kh\u00f4ng c\u1ea7n b\u1ed5 sung nh\u00e3n hu\u1ef3nh quang t\u1ed5ng h\u1ee3p ho\u1eb7c li\u00ean k\u1ebft n\u1ed9i sinh v\u1edbi protein hu\u1ef3nh quang.\u00a0Thay v\u00e0o \u0111\u00f3, k\u1ef9 thu\u1eadt n\u00e0y d\u1ef1a tr\u00ean c\u00e1c \u0111\u1eb7c t\u00ednh rung \u0111\u1ed9ng c\u1ee7a ph\u00e2n t\u1eed m\u1ee5c ti\u00eau v\u00e0 kh\u00f4ng y\u00eau c\u1ea7u lo\u00e0i n\u00e0y b\u1ecb k\u00edch th\u00edch \u0111i\u1ec7n t\u1eed b\u1edfi tia c\u1ef1c t\u00edm ho\u1eb7c \u00e1nh s\u00e1ng kh\u1ea3 ki\u1ebfn.\u00a0Trong th\u1ef1c t\u1ebf, c\u00e1c xung laze nhanh (pico gi\u00e2y ho\u1eb7c th\u1ea5p h\u01a1n) trong v\u00f9ng c\u1eadn h\u1ed3ng ngo\u1ea1i t\u1eeb hai ngu\u1ed3n \u0111\u01b0\u1ee3c h\u1ed9i t\u1ee5 v\u00e0o m\u1eabu v\u1eadt b\u1eb1ng v\u1eadt k\u00ednh hi\u1ec3n vi v\u00e0 \u0111\u01b0\u1ee3c qu\u00e9t raster trong c\u00e1c m\u1eb7t ph\u1eb3ng b\u00ean v\u00e0 tr\u1ee5c.\u00a0C\u00e1c xung \u0111\u01b0\u1ee3c ph\u00e2n t\u00e1ch theo t\u1ea7n s\u1ed1 b\u1eb1ng m\u1ed9t ch\u1ebf \u0111\u1ed9 dao \u0111\u1ed9ng ph\u00e2n t\u1eed \u0111\u00e3 ch\u1ecdn v\u00e0 t\u1ea1o ra m\u1ed9t ch\u00f9m tia m\u1edbi, c\u00f3 b\u01b0\u1edbc s\u00f3ng ng\u1eafn h\u01a1n c\u00e1c ch\u00f9m tia t\u1edbi, t\u1ea1i ti\u00eau \u0111i\u1ec3m v\u1eadt k\u00ednh.\u00a0Ch\u00f9m th\u1ee9 c\u1ea5p t\u1ea1o ra m\u1ed9t h\u1ed3 s\u01a1 n\u1ed3ng \u0111\u1ed9 c\u1ee7a c\u00e1c lo\u00e0i m\u1ee5c ti\u00eau v\u00e0 cho ph\u00e9p x\u00e2y d\u1ef1ng h\u00ecnh \u1ea3nh ba chi\u1ec1u c\u1ee7a m\u1eabu v\u1eadt.\u00a0B\u1edfi v\u00ec h\u1ea7u h\u1ebft c\u00e1c ph\u00e2n t\u1eed sinh h\u1ecdc \u0111\u01b0\u1ee3c t\u00ecm th\u1ea5y trong c\u00e1c t\u1ebf b\u00e0o s\u1ed1ng \u0111\u1ec1u c\u00f3 c\u00e1c kh\u1ed1i x\u00e2y d\u1ef1ng gi\u1ed1ng nhau n\u00ean r\u1ea5t kh\u00f3 \u0111\u1ec3 \u0111\u1ea1t \u0111\u01b0\u1ee3c t\u00ednh \u0111\u1eb7c hi\u1ec7u ph\u00e2n t\u1eed th\u1ef1c s\u1ef1 trong c\u00e1c th\u00ed nghi\u1ec7m CARS.\u00a0H\u01a1n n\u1eefa, CARS d\u1ef1a tr\u00ean s\u1ef1 c\u1ed9ng h\u01b0\u1edfng, ngh\u0129a l\u00e0 \u0111\u1ed9 nh\u1ea1y c\u1ee7a n\u00f3 b\u1ecb h\u1ea1n ch\u1ebf b\u1edfi th\u1ef1c t\u1ebf l\u00e0 ph\u1ea3i c\u00f3 h\u00e0ng ngh\u00ecn ph\u00e2n t\u1eed trong \u00e2m l\u01b0\u1ee3ng ti\u00eau \u0111i\u1ec3m \u0111\u1ec3 t\u1ea1o ra m\u1ee9c t\u00edn hi\u1ec7u \u0111\u00e1ng k\u1ec3.\u00a0Do \u0111\u00f3, CARS (\u1edf d\u1ea1ng hi\u1ec7n t\u1ea1i) ph\u1ea3i nh\u1eafm m\u1ee5c ti\u00eau v\u00e0o c\u00e1c ph\u00e2n t\u1eed phong ph\u00fa v\u00e0 c\u00f3 th\u1ec3 b\u1ecb h\u1ea1n ch\u1ebf d\u1ef1a v\u00e0o vi\u1ec7c ghi nh\u00e3n b\u00ean ngo\u00e0i (ch\u1eb3ng h\u1ea1n nh\u01b0 s\u1eed d\u1ee5ng c\u00e1c h\u1ee3p ch\u1ea5t kh\u1eed m\u00e0u) \u0111\u1ec3 cung c\u1ea5p t\u00ednh \u0111\u1eb7c hi\u1ec7u ph\u00e2n t\u1eed th\u1ef1c s\u1ef1.\u00a0r\u1ea5t kh\u00f3 \u0111\u1ec3 \u0111\u1ea1t \u0111\u01b0\u1ee3c t\u00ednh \u0111\u1eb7c hi\u1ec7u ph\u00e2n t\u1eed th\u1ef1c s\u1ef1 trong c\u00e1c th\u00ed nghi\u1ec7m CARS.\u00a0H\u01a1n n\u1eefa, CARS d\u1ef1a tr\u00ean s\u1ef1 c\u1ed9ng h\u01b0\u1edfng, ngh\u0129a l\u00e0 \u0111\u1ed9 nh\u1ea1y c\u1ee7a n\u00f3 b\u1ecb h\u1ea1n ch\u1ebf b\u1edfi th\u1ef1c t\u1ebf l\u00e0 ph\u1ea3i c\u00f3 h\u00e0ng ngh\u00ecn ph\u00e2n t\u1eed trong \u00e2m l\u01b0\u1ee3ng ti\u00eau \u0111i\u1ec3m \u0111\u1ec3 t\u1ea1o ra m\u1ee9c t\u00edn hi\u1ec7u \u0111\u00e1ng k\u1ec3.\u00a0Do \u0111\u00f3, CARS (\u1edf d\u1ea1ng hi\u1ec7n t\u1ea1i) ph\u1ea3i nh\u1eafm m\u1ee5c ti\u00eau v\u00e0o c\u00e1c ph\u00e2n t\u1eed phong ph\u00fa v\u00e0 c\u00f3 th\u1ec3 b\u1ecb h\u1ea1n ch\u1ebf d\u1ef1a v\u00e0o vi\u1ec7c ghi nh\u00e3n b\u00ean ngo\u00e0i (ch\u1eb3ng h\u1ea1n nh\u01b0 s\u1eed d\u1ee5ng c\u00e1c h\u1ee3p ch\u1ea5t kh\u1eed m\u00e0u) \u0111\u1ec3 cung c\u1ea5p t\u00ednh \u0111\u1eb7c hi\u1ec7u ph\u00e2n t\u1eed th\u1ef1c s\u1ef1.\u00a0r\u1ea5t kh\u00f3 \u0111\u1ec3 \u0111\u1ea1t \u0111\u01b0\u1ee3c t\u00ednh \u0111\u1eb7c hi\u1ec7u ph\u00e2n t\u1eed th\u1ef1c s\u1ef1 trong c\u00e1c th\u00ed nghi\u1ec7m CARS.\u00a0H\u01a1n n\u1eefa, CARS d\u1ef1a tr\u00ean s\u1ef1 c\u1ed9ng h\u01b0\u1edfng, ngh\u0129a l\u00e0 \u0111\u1ed9 nh\u1ea1y c\u1ee7a n\u00f3 b\u1ecb h\u1ea1n ch\u1ebf b\u1edfi th\u1ef1c t\u1ebf l\u00e0 ph\u1ea3i c\u00f3 h\u00e0ng ngh\u00ecn ph\u00e2n t\u1eed trong \u00e2m l\u01b0\u1ee3ng ti\u00eau \u0111i\u1ec3m \u0111\u1ec3 t\u1ea1o ra m\u1ee9c t\u00edn hi\u1ec7u \u0111\u00e1ng k\u1ec3.\u00a0Do \u0111\u00f3, CARS (\u1edf d\u1ea1ng hi\u1ec7n t\u1ea1i) ph\u1ea3i nh\u1eafm m\u1ee5c ti\u00eau v\u00e0o c\u00e1c ph\u00e2n t\u1eed phong ph\u00fa v\u00e0 c\u00f3 th\u1ec3 b\u1ecb h\u1ea1n ch\u1ebf d\u1ef1a v\u00e0o vi\u1ec7c ghi nh\u00e3n b\u00ean ngo\u00e0i (ch\u1eb3ng h\u1ea1n nh\u01b0 s\u1eed d\u1ee5ng c\u00e1c h\u1ee3p ch\u1ea5t kh\u1eed m\u00e0u) \u0111\u1ec3 cung c\u1ea5p t\u00ednh \u0111\u1eb7c hi\u1ec7u ph\u00e2n t\u1eed th\u1ef1c s\u1ef1.<\/p>\n<p><img loading=\"lazy\" decoding=\"async\" class=\"aligncenter\" src=\"https:\/\/zeiss-campus.magnet.fsu.edu\/articles\/livecellimaging\/images\/techniquesfigure10.jpg\" width=\"386\" height=\"314\" border=\"0\" \/>K\u00ednh hi\u1ec3n vi <b>t\u1ea1o s\u00f3ng h\u00e0i <\/b>l\u00e0 m\u1ed9t k\u1ef9 thu\u1eadt m\u1edbi n\u1ed5i m\u00e0 cu\u1ed1i c\u00f9ng c\u00f3 th\u1ec3 \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng r\u1ed9ng r\u00e3i trong h\u00ecnh \u1ea3nh t\u1ebf b\u00e0o s\u1ed1ng.\u00a0S\u1ef1 t\u1ea1o ra s\u00f3ng h\u00e0i x\u1ea3y ra khi m\u1ed9t s\u1ef1 ki\u1ec7n k\u00edch th\u00edch quang h\u1ecdc li\u00ean quan \u0111\u1ebfn hai ho\u1eb7c nhi\u1ec1u photon \u1edf m\u1ed9t t\u1ea7n s\u1ed1 c\u1ee5 th\u1ec3 d\u1eabn \u0111\u1ebfn s\u1ef1 ph\u00e1t x\u1ea1 h\u1ee3p t\u00e1c \u1edf nhi\u1ec1u s\u00f3ng h\u00e0i (ch\u1ee7 y\u1ebfu l\u00e0 s\u00f3ng h\u00e0i th\u1ee9 hai v\u00e0 th\u1ee9 ba) m\u00e0 kh\u00f4ng c\u00f3 s\u1ef1 h\u1ea5p th\u1ee5 c\u1ee7a c\u00e1c photon.\u00a0Vi\u1ec7c t\u1ea1o ra c\u00e1c t\u1ea7n s\u1ed1 s\u00f3ng h\u00e0i v\u1ec1 c\u01a1 b\u1ea3n l\u00e0 m\u1ed9t qu\u00e1 tr\u00ecnh t\u00e1n x\u1ea1 phi tuy\u1ebfn t\u00ednh t\u1ea1o ra b\u01b0\u1edbc s\u00f3ng photon ph\u00e1t ra g\u1ea5p \u0111\u00f4i t\u1ea7n s\u1ed1 ho\u1eb7c m\u1ed9t n\u1eeda b\u01b0\u1edbc s\u00f3ng (\u0111\u1ed1i v\u1edbi s\u1ef1 t\u1ea1o s\u00f3ng h\u00e0i th\u1ee9 hai) c\u1ee7a \u00e1nh s\u00e1ng t\u1edbi.\u00a0Trong k\u00ednh hi\u1ec3n vi quang h\u1ecdc, c\u00e1c m\u1eabu v\u1eadt trong su\u1ed1t thi\u1ebfu t\u00ednh \u0111\u1ed1i x\u1ee9ng (ch\u1eb3ng h\u1ea1n nh\u01b0 t\u1ebf b\u00e0o s\u1ed1ng) l\u00e0 \u1ee9ng c\u1eed vi\u00ean l\u00fd t\u01b0\u1edfng \u0111\u1ec3 ch\u1ee5p \u1ea3nh b\u1eb1ng c\u00e1c k\u1ef9 thu\u1eadt t\u1ea1o s\u00f3ng h\u00e0i.\u00a0Kh\u00f4ng gi\u1ed1ng nh\u01b0 t\u00ecnh hu\u1ed1ng v\u1edbi c\u00e1c \u0111\u1ea7u d\u00f2 \u0111i\u1ec3n h\u00ecnh v\u00e0 k\u1ef9 thu\u1eadt chi\u1ebfu s\u00e1ng b\u1eb1ng k\u00ednh hi\u1ec3n vi hu\u1ef3nh quang truy\u1ec1n th\u1ed1ng,\u00a0thay \u0111\u1ed5i b\u01b0\u1edbc s\u00f3ng chi\u1ebfu s\u00e1ng k\u00edch th\u00edch t\u1ea1o ra s\u1ef1 thay \u0111\u1ed5i t\u01b0\u01a1ng \u1ee9ng trong b\u01b0\u1edbc s\u00f3ng ph\u00e1t x\u1ea1.\u00a0Ngo\u00e0i ra, \u00e1nh s\u00e1ng ph\u00e1t ra l\u00e0 nh\u1ea5t qu\u00e1n v\u00e0 gi\u1eef l\u1ea1i th\u00f4ng tin pha v\u1ec1 m\u1eabu v\u1eadt.<\/p>\n<p>\u0110\u1ec3 c\u00f3 \u0111\u1ed9 ph\u00e2n gi\u1ea3i quang h\u1ecdc c\u1ef1c cao, k\u00ednh hi\u1ec3n vi quang h\u1ecdc\u00a0<b>qu\u00e9t tr\u01b0\u1eddng g\u1ea7n<\/b>(<b>NSOM<\/b>) hi\u1ec7n l\u00e0 k\u1ef9 thu\u1eadt quang t\u1eed \u0111\u01b0\u1ee3c l\u1ef1a ch\u1ecdn.\u00a0H\u00ecnh \u1ea3nh tr\u01b0\u1eddng g\u1ea7n x\u1ea3y ra khi m\u1ed9t \u0111\u1ea7u d\u00f2 quang h\u1ecdc c\u00f3 k\u00edch th\u01b0\u1edbc micron ph\u1ee5 \u0111\u01b0\u1ee3c \u0111\u1eb7t c\u00e1ch m\u1eabu m\u1ed9t kho\u1ea3ng r\u1ea5t ng\u1eafn v\u00e0 \u00e1nh s\u00e1ng \u0111\u01b0\u1ee3c truy\u1ec1n qua m\u1ed9t l\u1ed7 nh\u1ecf \u1edf \u0111\u1ea7u c\u1ee7a \u0111\u1ea7u d\u00f2 n\u00e0y.\u00a0Tr\u01b0\u1eddng g\u1ea7n \u0111\u01b0\u1ee3c \u0111\u1ecbnh ngh\u0129a l\u00e0 v\u00f9ng ph\u00eda tr\u00ean m\u1ed9t b\u1ec1 m\u1eb7t c\u00f3 k\u00edch th\u01b0\u1edbc nh\u1ecf h\u01a1n m\u1ed9t b\u01b0\u1edbc s\u00f3ng \u00e1nh s\u00e1ng t\u1edbi tr\u00ean b\u1ec1 m\u1eb7t \u0111\u00f3.\u00a0Trong v\u00f9ng tr\u01b0\u1eddng g\u1ea7n, \u00e1nh s\u00e1ng bay h\u01a1i kh\u00f4ng b\u1ecb gi\u1edbi h\u1ea1n nhi\u1ec5u x\u1ea1 v\u00e0 c\u00f3 th\u1ec3 ph\u00e2n gi\u1ea3i kh\u00f4ng gian nanomet.\u00a0Hi\u1ec7n t\u01b0\u1ee3ng n\u00e0y cho ph\u00e9p ch\u1ee5p \u1ea3nh v\u00e0 quang ph\u1ed5 gi\u1edbi h\u1ea1n kh\u00f4ng nhi\u1ec5u x\u1ea1 c\u1ee7a m\u1ed9t m\u1eabu m\u00e0 ph\u01b0\u01a1ng ph\u00e1p ch\u1ee5p \u1ea3nh quang h\u1ecdc th\u00f4ng th\u01b0\u1eddng kh\u00f4ng th\u1ec3 th\u1ef1c hi\u1ec7n \u0111\u01b0\u1ee3c.\u00a0M\u1eb7c d\u00f9 hi\u1ec7n nay hi\u1ebfm khi \u0111\u01b0\u1ee3c \u00e1p d\u1ee5ng cho h\u00ecnh \u1ea3nh t\u1ebf b\u00e0o s\u1ed1ng, c\u00e1c k\u1ef9 thu\u1eadt li\u00ean quan nh\u01b0 k\u00ednh hi\u1ec3n vi\u00a0<b>l\u1ef1c nguy\u00ean t\u1eed<\/b>\u00a0\u0111ang \u0111\u01b0\u1ee3c kh\u00e1m ph\u00e1 \u0111\u1ec3 ki\u1ec3m tra b\u1ec1 m\u1eb7t c\u1ee7a c\u00e1c t\u1ebf b\u00e0o s\u1ed1ng.<\/p>\n<p>Hi\u1ec3n th\u1ecb \u0111\u1ed9 ph\u00e2n gi\u1ea3i kh\u00f4ng gian v\u01b0\u1ee3t xa gi\u1edbi h\u1ea1n nhi\u1ec5u x\u1ea1,\u00a0k\u00ednh\u00a0hi\u1ec3n vi<b> suy gi\u1ea3m ph\u00e1t x\u1ea1 k\u00edch th\u00edch (\u00a0<\/b><b>STED<\/b>) l\u00e0 m\u1ed9t k\u1ef9 thu\u1eadt si\u00eau ph\u00e2n gi\u1ea3i m\u1edbi n\u1ed5i s\u1eed d\u1ee5ng ch\u00f9m suy gi\u1ea3m h\u00ecnh b\u00e1nh r\u00e1n bao quanh ch\u00f9m k\u00edch th\u00edch nh\u1ecf h\u01a1n \u0111\u1ec3 \u0111\u1ea1t \u0111\u01b0\u1ee3c \u0111\u1ed9 ph\u00e2n gi\u1ea3i tr\u1ee5c th\u1ea5p h\u01a1n 50 nanomet. K\u1ef9 thu\u1eadt n\u00e0y d\u1ef1a v\u00e0o vi\u1ec7c \u1ee9c ch\u1ebf s\u1ef1 ph\u00e1t hu\u1ef3nh quang c\u1ee7a c\u00e1c ph\u00e2n t\u1eed b\u1ecb k\u00edch th\u00edch \u1edf ngo\u1ea1i vi c\u1ee7a ti\u00eau \u0111i\u1ec3m qu\u00e9t laze b\u1eb1ng c\u00e1ch s\u1eed d\u1ee5ng c\u00e1c xung laze \u0111\u01b0\u1ee3c \u0111\u1ed3ng b\u1ed9 h\u00f3a \u0111\u1ec3 k\u00edch th\u00edch c\u00e1c ch\u1ea5t hu\u1ef3nh quang v\u00e0 c\u00e1c xung STED tr\u00f2n \u0111\u01b0\u1ee3c ph\u1ed1i h\u1ee3p kh\u00f4ng gian \u0111\u1ec3 l\u00e0m gi\u1ea3m ph\u00e1t x\u1ea1. K\u1ebft qu\u1ea3 l\u00e0 hu\u1ef3nh quang b\u1ecb \u1ee9c ch\u1ebf \u1edf ngo\u1ea1i vi c\u1ee7a v\u1ebft, nh\u01b0ng kh\u00f4ng ph\u1ea3i \u1edf trung t\u00e2m, do \u0111\u00f3 l\u00e0m gi\u1ea3m \u0111\u00e1ng k\u1ec3 k\u00edch th\u01b0\u1edbc v\u1ebft hu\u1ef3nh quang c\u00f9ng v\u1edbi s\u1ef1 gia t\u0103ng \u0111\u1ed9 ph\u00e2n gi\u1ea3i. STED \u0111\u00e3 \u0111\u01b0\u1ee3c ch\u1ee9ng minh l\u00e0 m\u1ed9t c\u00f4ng c\u1ee5 h\u1eefu \u00edch \u0111\u1ec3 ki\u1ec3m tra c\u00e1c t\u1ebf b\u00e0o s\u1ed1ng \u1edf \u0111\u1ed9 ph\u00e2n gi\u1ea3i cao. C\u00e1c k\u1ef9 thu\u1eadt si\u00eau ph\u00e2n gi\u1ea3i m\u1edbi n\u1ed5i kh\u00e1c, ch\u1eb3ng h\u1ea1n nh\u01b0 k\u00ednh hi\u1ec3n vi \u0111\u1ecbnh v\u1ecb quang h\u00f3a (<b>PALM<\/b>) v\u00e0 k\u00ednh hi\u1ec3n vi chi\u1ebfu s\u00e1ng c\u00f3 c\u1ea5u tr\u00fac (\u00a0<b>SIM<\/b>\u00a0) c\u00f3 th\u1ec3 s\u1ebd tr\u1edf th\u00e0nh c\u00f4ng c\u1ee5 thi\u1ebft y\u1ebfu trong ch\u1ee5p \u1ea3nh t\u1ebf b\u00e0o s\u1ed1ng trong t\u01b0\u01a1ng lai g\u1ea7n.<\/p>\n<h2><span class=\"ez-toc-section\" id=\"Ket_luan\"><\/span><b>K\u1ebft lu\u1eadn<\/b><span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p>Vi\u1ec7c s\u1eed d\u1ee5ng ng\u00e0y c\u00e0ng nhi\u1ec1u c\u00e1c protein hu\u1ef3nh quang \u0111\u01b0\u1ee3c m\u00e3 h\u00f3a di truy\u1ec1n v\u00e0 c\u00e1c fluorophores t\u1ed5ng h\u1ee3p ti\u00ean ti\u1ebfn \u0111\u1ec3 ch\u1ee5p \u1ea3nh t\u1ebf b\u00e0o s\u1ed1ng \u0111\u00e3 m\u1edf c\u01a1 h\u1ed9i cho c\u00e1c ph\u01b0\u01a1ng th\u1ee9c quang h\u1ecdc h\u1eefu \u00edch. Nh\u00e0 k\u00ednh hi\u1ec3n vi hi\u1ec7n c\u00f3 \u0111\u1ea7y \u0111\u1ee7 c\u00e1c c\u00f4ng c\u1ee5 \u0111\u1ec3 xem v\u00e0 ghi l\u1ea1i d\u1eef li\u1ec7u h\u00ecnh \u1ea3nh c\u1ee7a c\u00e1c qu\u00e1 tr\u00ecnh x\u1ea3y ra trong m\u1ed9t kho\u1ea3ng th\u1eddi gian d\u00e0i v\u00e0 \u1edf nhi\u1ec1u \u0111\u1ed9 ph\u00e2n gi\u1ea3i. C\u00e1c s\u1ef1 ki\u1ec7n d\u1ec5 d\u00e0ng \u0111\u01b0\u1ee3c quan s\u00e1t v\u00e0 ghi l\u1ea1i b\u1eb1ng k\u00ednh hi\u1ec3n vi \u0111\u1ed3ng ti\u00eau qu\u00e9t laze, trong khi c\u00f3 th\u1ec3 truy c\u1eadp nhanh h\u01a1n th\u00f4ng qua vi\u1ec7c s\u1eed d\u1ee5ng c\u00e1c k\u1ef9 thu\u1eadt \u0111\u0129a quay. Ngo\u00e0i ra, k\u00ednh hi\u1ec3n vi \u0111a \u0111i\u1ec3m cho ph\u00e9p ch\u1ee5p \u1ea3nh s\u00e2u b\u00ean trong c\u00e1c m\u00f4 d\u00e0y v\u00e0 k\u1ef9 thu\u1eadt ph\u1ea3n x\u1ea1 to\u00e0n ph\u1ea7n b\u00ean trong c\u00f3 th\u1ec3 th\u0103m d\u00f2 b\u1ec1 m\u1eb7t m\u00e0ng v\u1edbi \u0111\u1ed9 ch\u00ednh x\u00e1c \u0111\u1ed3ng ti\u00eau. Ph\u01b0\u01a1ng ph\u00e1p hu\u1ef3nh quang ti\u00ean ti\u1ebfn, ch\u1eb3ng h\u1ea1n nh\u01b0 FRET, FLIM, FRAP, FCS, FSM, SIM, PALM v\u00e0 STED, c\u00f3 th\u1ec3 \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng \u0111\u1ec3 theo d\u00f5i c\u00e1c t\u01b0\u01a1ng t\u00e1c protein-protein, th\u01b0\u1eddng \u1edf \u0111\u1ed9 ph\u00e2n gi\u1ea3i l\u1edbn h\u01a1n \u0111\u1ed9 ph\u00e2n gi\u1ea3i cho ph\u00e9p.<\/p>\n<p><em>Ngu\u1ed3n: <a href=\"https:\/\/zeiss-campus.magnet.fsu.edu\/articles\/livecellimaging\/techniques.html\" target=\"_blank\" rel=\"noopener\">https:\/\/zeiss-campus.magnet.fsu.edu\/articles\/livecellimaging\/techniques.html<\/a><\/em><\/p>\n<p><strong>C\u00f4ng ty Minh Khang l\u00e0 nh\u00e0 ph\u00e2n ph\u1ed1i \u0111\u1ed9c quy\u1ec1n th\u1ecb tr\u01b0\u1eddng mi\u1ec1n Nam ph\u00e2n kh\u00fac k\u00ednh hi\u1ec3n vi h\u00e3ng\u00a0<a href=\"https:\/\/vospan.com\/product-brands\/zeiss-vi\/\">Carl ZEISS<\/a>.<\/strong><\/p>\n","protected":false},"excerpt":{"rendered":"<p>Gi\u1edbi thi\u1ec7u Theo d\u00f5i h\u00ecnh \u1ea3nh chuy\u1ec3n \u0111\u1ed9ng c\u1ee7a t\u1ebf b\u00e0o s\u1ed1ng theo ph\u01b0\u01a1ng ph\u00e1p truy\u1ec1n th\u1ed1ng b\u1eb1ng c\u00e1ch s\u1eed d\u1ee5ng c\u00e1c k\u1ef9 thu\u1eadt t\u0103ng c\u01b0\u1eddng \u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n ph\u1ed5 bi\u1ebfn bao g\u1ed3m tr\u01b0\u1eddng s\u00e1ng, \u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n nhi\u1ec5u vi sai (DIC), \u0111\u1ed9 t\u01b0\u01a1ng ph\u1ea3n \u0111i\u1ec1u bi\u1ebfn Hoffman (HMC), t\u01b0\u01a1ng ph\u1ea3n pha v\u00e0 hu\u1ef3nh quang&#8230;<\/p>\n","protected":false},"author":3,"featured_media":9406,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"footnotes":""},"categories":[40],"tags":[],"class_list":["post-9405","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-tin-tuc"],"acf":[],"_links":{"self":[{"href":"https:\/\/vospan.com\/wp-json\/wp\/v2\/posts\/9405"}],"collection":[{"href":"https:\/\/vospan.com\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/vospan.com\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/vospan.com\/wp-json\/wp\/v2\/users\/3"}],"replies":[{"embeddable":true,"href":"https:\/\/vospan.com\/wp-json\/wp\/v2\/comments?post=9405"}],"version-history":[{"count":4,"href":"https:\/\/vospan.com\/wp-json\/wp\/v2\/posts\/9405\/revisions"}],"predecessor-version":[{"id":9851,"href":"https:\/\/vospan.com\/wp-json\/wp\/v2\/posts\/9405\/revisions\/9851"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/vospan.com\/wp-json\/wp\/v2\/media\/9406"}],"wp:attachment":[{"href":"https:\/\/vospan.com\/wp-json\/wp\/v2\/media?parent=9405"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/vospan.com\/wp-json\/wp\/v2\/categories?post=9405"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/vospan.com\/wp-json\/wp\/v2\/tags?post=9405"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}