[Histonet] GI biopsy H&E staining

Annie T. Walton AWalton <@t> oima.org
Fri May 10 13:02:30 CDT 2013

Hello all,

Our Medical Director would like the goblet cells on our GI biopsies to stain a darker blue. I hand stain using the following protocol:

 *   Xylene, 3 changes                                                                            3 minutes each
 *   100% Alcohol, 2 changes                                                               1 minute each
 *   95% Alcohol                                                                                        1 minute
 *   Tap water                                                                                            rinse until water runs off evenly
 *   Harris Hematoxylin                                                                          5 minutes
 *   Water wash                                                                                        rinse until water runs clear
 *   Clarifier 2                                                                                             5 seconds
 *   Running water                                                                                  30 seconds
 *   Bluing Solution                                                                                  1 minute
 *   Tap water                                                                                            rinse well
 *   95% Alcohol                                                                                        30 seconds
 *   Eosin Y                                                                                                  1 minute
 *   100% Alcohol, 2 changes                                                               30 seconds
 *   100% Alcohol, 2 changes                                                               1 minute each
 *   Xylene, 3 changes                                                                            1 minute each

I have recently increased the hematoxylin time from 4 to 5 minutes and decreased the time in clarifier from 25 to 5 seconds so the hematoxylin doesn't get washed out as much, but it is not good enough. I also switched from 70% alcohol to 95% before eosin and increased the time in eosin from 45 seconds to 1 minute to provide a greater contrast in hopes that that would work, but the goblet cells are still not staining dark enough. Any suggestions would greatly be appreciated!


Annie Walton, HTL (ASCP)
Histology Technician
Overlake Internal Medicine Associates Histology Lab
awalton <@t> oima.org

DISCLAIMER: This message is confidential, intended only for the named recipient(s) and may contain information that is privileged or exempt from disclosure under applicable law. If you are not the intended recipient(s), you are notified that the dissemination, distribution or copying of this information is strictly prohibited. If you received this message in error, please notify the sender then delete this message.

More information about the Histonet mailing list