| Autor: |
Jelmer, Willems, Manon, Westra, Harold D, MacGillavry |
| Rok vydání: |
2022 |
| Předmět: |
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| Zdroj: |
Methods in molecular biology (Clifton, N.J.). 2440 |
| ISSN: |
1940-6029 |
| Popis: |
Over the past years several forms of superresolution fluorescence microscopy have been developed that offer the possibility to study cellular structures and protein distribution at a resolution well below the diffraction limit of conventional fluorescence microscopy (200 nm). A particularly powerful superresolution technique is single-molecule localization microscopy (SMLM). SMLM enables the quantitative investigation of subcellular protein distribution at a spatial resolution up to tenfold higher than conventional imaging, even in live cells. Not surprisingly, SMLM has therefore been used in many applications in biology, including neuroscience. This chapter provides a step-by-step SMLM protocol to visualize the nanoscale organization of endogenous proteins in dissociated neurons but can be extended to image other adherent cultured cells. We outline a number of methods to visualize endogenous proteins in neurons for live-cell and fixed application, including immunolabeling, the use of intrabodies for live-cell SMLM, and endogenous tagging using CRISPR/Cas9. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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