期刊
ANALYTICAL CHEMISTRY
卷 93, 期 25, 页码 8698-8703出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.1c01702
关键词
-
资金
- Natural Science Foundation of China [61475059, 61890950]
- 973 programs [2015CB755603]
Immunofluorescence chemical sectioning (IF-CS) utilizes complexation reactions to provide an off/on mechanism for Alexa dyes, allowing CS imaging of IF labeled tissues, which solves the conflict in traditional imaging methods. IF-CS demonstrates its utility in wide-field imaging of large immunolabeled tissues with three-dimensional submicron resolution, greatly facilitating systematic studies of refined subcellular architectures in intact biological systems.
Immunofluorescence (IF) is a powerful investigative tool in biological research and medical diagnosis, whereas conventional imaging methods are always conflict between speed, contrast/resolution, and specimen volume. Chemical sectioning (CS) is an effective method to overcome the conflict, which works by chemically manipulating the off/on state of fluorescent materials and turning on only the extremely superficial surface fluorescence of tissues to realize the sectioning capacity of wide-field imaging. However, the current mechanism of CS is only applicable to samples labeled with pH-sensitive fluorescent proteins and still cannot fulfill samples immunolabeled with frequently used commercial fluorescent dyes. Here, immunofluorescence chemical sectioning (IF-CS) is described to present an off/on mechanism for Alexa dyes by complexation reactions, allowing CS imaging of IF labeled tissues. IF-CS enables IF freeing from out-of-focus interference in wide-field imaging and satisfying with multicolor imaging. IF-CS demonstrates the utility of the 3D submicron-resolution imaging of large immunolabeled tissues on the wide-field block-face system. IF-CS may remarkably facilitate systematic studies of refined subcellular architectures of endogenous proteins in intact biological systems.
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