[Histonet] Re: Harris hematoxylin weak staining
TJJ <@t> stowers.org
Mon Sep 21 08:57:13 CDT 2009
There are several strategies you can use to improve your hematoxylin staining.
1 - use distilled or DI water before putting your slides into the stain. Sometimes the chemicals/minerals/metals in the tap water can weaken your stain.
2 - replace your stains when they start showing signs of weakening, or routinely in order to keep the quality even.
3 - try using more than one bucket/dish of hematoxylin and split the total staining time between them. Then you can rotate the last one up to the first position, and put a new change in the second station. This might help keep your staining more consistent.
4 - does your tap water seem to contain a high level of chlorine? If so, you can use a bluing agent to blue your stained slides, and then use DI or distilled water rinses.
5 - do not linger in the decolorizing acid alcohol. Usually one quick dip followed by an immediate plunge into water is all you need.
6 - some have reported that their eosin, if the pH drops too low, can leach out some of the hematoxylin. According to Carson, the ideal pH of eosin is between 4.6 and 5.
7 - what thickness is your tissue? 3 microns thin tissue needs longer in the stain solution than 5 microns thin tissue.
There may be others that folks will chime in with, but these are the most obvious ones to me.
Teri Johnson, HT(ASCP)QIHC
Managing Director Histology Facility
Stowers Institute for Medical Research
1000 E. 50th St.
Kansas City, MO 64110
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