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Expansion Pathology: A Method for High-resolution Optical Imaging of Clinical Tissue Samples
Overview
In this video, we describe a method called expansion pathology, a variant of expansion microscopy, wherein the clinical tissue specimens are chemically expanded using a polymer network for microscopic examination of nanoscale structures.
Protocol
1. In Situ Polymerization of Specimens
- Incubate the specimen in an anchoring solution.
- Prepare the anchoring solution (typically 250 µL is enough to cover the tissue section) by diluting the AcX stock solution in 1x PBS to a concentration of 0.03 mg/mL for samples fixed with non-aldehyde fixatives or 0.1 mg/mL for samples fixed with aldehyde fixatives, which have fewer free amines available to react with AcX.
- Place the slide in a 100 mm Petri dish and pipette the a.......
Representative Results
Figure 1: Gelation chamber for pathology samples. (A) Two spacers, such as two pieces of #1.5 cover glass cut with a diamond knife, are placed on either side of the tissue after incubating the sample in gelling solution at 4 °C. The spacers should be thicker than the tissue slices, to prevent compression of the sample. (B) A lid, such a.......
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Source: Klimas, A. et al. Nanoscopic Imaging of Human Tissue Sections via Physical and Isotropic Expansion. J. Vis. Exp. (2019)
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