Yao Longfang, Zhang Li, Fei Yiyan, Chen Liwen, Mi Lan, Ma Jiong
Department of Optical Science and Engineering, Shanghai Engineering Research Center of Ultra-Precision Optical Manufacturing, Key Laboratory of Micro and Nano Photonic Structures (Ministry of Education), Green Photoelectron Platform, Fudan University, Shanghai, China.
Shanghai Engineering Research Center of Industrial Microorganisms, The Multiscale Research Institute of Complex Systems, School of Life Sciences, Fudan University, Shanghai, China.
Front Chem. 2021 Apr 30;9:640519. doi: 10.3389/fchem.2021.640519. eCollection 2021.
Expansion super-resolution technology is a new technology developed in recent years. It anchors the dye on the hydrogel and the dye expands with the expansion of the hydrogel so that a super-resolution map can be obtained under an ordinary microscope. However, by labeling the target protein with a first antibody and secondary antibody, the distance between the fluorescent group and the actual target protein is greatly increased. Although fluorescent proteins can also be used for expansion super-resolution to reduce this effect, the fluorescent protein is often destroyed during sample preparation. To solve this problem, we developed a novel label system for expansion microscopy, based on a DNA oligostrand linked with a fluorescent dye, acrylamide group (linker), and benzoylguanine (BG, a small substrate molecule for SNAP-tag). This protocol greatly reduced the error between the position of fluorescent group and the actual target protein, and also reduced loss of the fluorescent group during sample preparation.
膨胀超分辨率技术是近年来发展起来的一项新技术。它将染料锚定在水凝胶上，随着水凝胶的膨胀，染料也随之膨胀，从而在普通显微镜下就能获得超分辨率图像。然而，通过用一抗和二抗标记目标蛋白，荧光基团与实际目标蛋白之间的距离会大大增加。虽然荧光蛋白也可用于膨胀超分辨率以减少这种影响，但在样品制备过程中荧光蛋白常常会被破坏。为了解决这个问题，我们基于与荧光染料、丙烯酰胺基团（连接体）和苯甲酰鸟嘌呤（BG，一种用于SNAP标签的小分子底物）相连的DNA寡链，开发了一种用于膨胀显微镜的新型标记系统。该方案大大减少了荧光基团位置与实际目标蛋白之间的误差，同时也减少了样品制备过程中荧光基团的损失。
