Chapter title |
Expansion Microscopy of Ciliary Proteins.
|
---|---|
Book title |
Cilia
|
Published in |
Methods in molecular biology, January 2024
|
DOI | 10.1007/978-1-0716-3507-0_4 |
Pubmed ID | |
Book ISBNs |
978-1-07-163506-3, 978-1-07-163507-0
|
Authors |
Park, Sohyeon, Shi, Xiaoyu |
Abstract |
Label-retention expansion microscopy (LR-ExM) is a sample preparation technique, which embeds the cells or tissues in a swellable hydrogel and expands the sample so that one can achieve a high resolution with any conventional fluorescence microscopes. Fluorescence loss during polymerization and protein denaturation have been a major limitation of standard expansion microscopy. To minimize fluorescence loss, LR-ExM uses trifunctional anchors, which can survive from polymerization and denaturation, and then introduce fluorophores after expansion. By using LR-ExM, one can study the structure of primary cilia at molecular-scale resolution with a much higher signal-to-noise ratio, compared with previously introduced expansion microscopy methods. In this chapter, we describe a detailed procedure showing how LR-ExM is used to study ciliary proteins. |
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