Protein-retention expansion microscopy for visualizing subcellular organelles in fixed brain tissue.

ABSTRACT:

Background

Protein expansion microscopy (proExM) is a powerful technique that crosslinks proteins to a swellable hydrogel to physically expand and optically clear biological samples. The resulting increased resolution (~70 nm) and physical separation of labeled proteins make it an attractive tool for studying the localization of subcellular organelles in densely packed tissues, such as the brain. However, the digestion and expansion process greatly reduce fluorescence signals making it necessary to optimize ExM conditions per sample for specific end goals.

New method

Here we compare the staining and digestion conditions of existing proExM workflows to identify the optimal protocol for visualizing subcellular organelles (mitochondria and the Golgi apparatus) within reporter-l

SUBMITTER: Campbell LA 

PROVIDER: S-EPMC8370715 | biostudies-literature | 2021 Sep

REPOSITORIES: biostudies-literature