A Guide to Build a Highly Inclined Swept Tile Microscope for Extended Field-of-view Single-molecule Imaging
| Autor: | Kyu Young Han, Jeffrey B. Oleske, Chun-Hung Weng, Jialei Tang |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Physics
Microscopy Microscope General Immunology and Microbiology business.industry Confocal General Chemical Engineering General Neuroscience Oxides Single Molecule Imaging Signal General Biochemistry Genetics and Molecular Biology law.invention Mirror galvanometer Optics Optical microscope Semiconductors law business Beam (structure) Lenses |
| Zdroj: | Journal of visualized experiments : JoVE. (146) |
| ISSN: | 1940-087X |
| Popis: | Single-molecule imaging has greatly advanced our understanding of molecular mechanisms in biological studies. However, it has been challenging to obtain large field-of-view, high-contrast images in thick cells and tissues. Here, we introduce highly inclined swept tile (HIST) microscopy that overcomes this problem. A pair of cylindrical lenses was implemented to generate an elongated excitation beam that was scanned over a large imaging area via a fast galvo mirror. A 4f configuration was used to position optical components. A scientific complementary metal-oxide semiconductor camera detected the fluorescence signal and blocked the out-of-focus background with a dynamic confocal slit synchronized with the beam sweeping. We present a step-by-step instruction on building the HIST microscope with all basic components. |
| Databáze: | OpenAIRE |
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