Processing slides for LCM (Version 1.0)

Version

1.0

Notice

This page is the corresponding protocol tomestone page generated as part of the ATLAS-D2K shutdown in July 2025. Many links on this page may be broken.

Authors

Steve Potter

Keywords

[‘Laser Capture Microdissection’]

Subjects

[‘Isolation, purification and separation’, ‘Gene expression analysis’]

Release Date

2017-08-07

Abstract

Processing slides for LCM protocol by the Potter Group

Procedure

1. Remove slides from –80°C freezer, store temporarily on dry ice, air dry on metal slide warmer turned off or set to about 30 to 35°C, for 3 min.
2. Fix two minutes. 1:1 mix of acetone 75% ethanol gives a nice compromise, with good section sticking and reasonable histology, and good lectin staining.
3. Rinse 2 min in 1/10X PBS, ice cold, to dissolve off some OCT. The 1/10 PBS is made by diluting 1X PBS ten fold with sterile autoclaved super water.
4. Stain about 6 min in lectin, on ice. For PNA use 5 μl per ml of 1/10 PBS. Stain on ice.
5. Rinse 1/10 PBS, ice cold. Two gentle 10 sec dips and then 3 min.
6. Dehydrate, in ethanol, 75%, 95%, 100%, 10-15 sec each, with 2-3 dips, and then another 100% one minute.
7. Xylene 1.5 min with three dips.
8. Xylene 2 min with two dips.
9. Air dry about 2-4 min.

Consortium

GenitoUrinary Development Molecular Anatomy Project (GUDMAP) Consortium