Chen Lab, 2009

H & E STAINING
This procedure is performed with the slides in glass staining racks and the solutions in square glass staining jars; seal the jars with parafilm while not in use; solutions should be made fresh every few weeks. Dip the glass racks with slides (from cryosection or -80 0C) serially in the following jars: - 95% EtOH, 2 minutes - 70% EtOH, 2 minutes - Running water, 5 minutes [Note: keep regular tap water running in a big tank in the sink and place the glass rack in the tank.] - Gills Hematoxylin, 3 minutes [Note: 3 min yields optimal staining, longer than that, background starts to show up.] - Running water, 5 minutes - Scotts solution, 3 min - Running water, 5 minutes - Eosin 1x, 1.5 minute [Note: 1.5 min yields optimal staining; dilute 2x stock 1:1 with 70% ethanol before staining.] - 70% EtOH, 30 seconds - 95% EtOH, 30 seconds - 100% EtOH, 30 seconds - Xylene, 5 min [Note: perform in the chemical hood.] - Mount with Permount (Fisher) and coverslip [Note: wait a few min for Xylene to dry off before mounting.] Solutions Scotts solution: 2g Sodium Bicarnobate 20g MgSO4 1 liter water 2x Eosin (dilute 1:1 with 70% ethanol before staining) 5g Eosin 2.5mL acetic acid 500mL 70% ethanol Gills Hematoxylin and Xylene are purchased from Fisher.