학술논문

Universal Molecular Retention with 11-Fold Expansion Microscopy
Document Type
research-article
Author
Source
Journal of Visualized Experiments. (200)
Subject
This Month in JoVE
Issue 200
Expansion microscopy
fluorescence microscopy
methacrolein
magnify
nanoscale imaging
super-resolution imaging
pathology
expansion pathology
immunohistochemistry
post-expansion staining
Language
English
ISSN
1940-087X
Abstract
The nanoscale imaging of biological specimens can improve the understanding of disease pathogenesis. In recent years, expansion microscopy (ExM) has been demonstrated to be an effective and low-cost alternative to optical super-resolution microscopy. However, it has been limited by the need for specific and often custom anchoring agents to retain different biomolecule classes within the gel and by difficulties with expanding standard clinical sample formats, such as formalin-fixed paraffin-embedded tissue, especially if larger expansion factors or preserved protein epitopes are desired. Here, we describe Magnify, a new ExM method for robust expansion up to 11-fold in a wide array of tissue types. By using methacrolein as the chemical anchor between the tissue and gel, Magnify retains multiple biomolecules, such as proteins, lipids, and nucleic acids, within the gel, thus allowing the broad nanoscale imaging of tissues on conventional optical microscopes. This protocol describes best practices to ensure robust and crack-free tissue expansion, as well as tips for handling and imaging highly expanded gels.