The Special Stain Kit Challenge

By: Ron Hightower, Director of Quality Assurance, American Master*Tech Scientific, Inc. Lodi, CA.

	Although the Hematoxylin and Eosin (H&E) Stain is the most often performed histology stain, many of the so-called "special" histochemical stains are done so frequently that they are considered "routine" stains by most histotechnologists. Stains such as Periodic Acid - Schiff's, Mucicarmine, Modified G.M.S., Reticulum, Gram, Steiner, and Masson's Trichrome are used extensively in clinical pathology and are increasingly utilized in medical research projects. In September 1983, American Histology Reagent Company, precursor to American Master*Tech Scientific, Inc., introduced a G.M.S. Stain Kit, one of the first commercially available histochemical stain kits. By 1989, there were 16 special stain kits offered in the A.H.R.C. Product Catalog; in this edition of the Master*Tech Magazine, 40 special stain kits are available! This wide variety of histochemical special stain kits is the result of over 25 years of service to histotechnologists in the United States and throughout the world. As part of our ongoing commitment to provide you with special stain kits that meet your requirements for utility, stain quality, and economy, we performed a study comparing Master*Tech G.M.S., Muci Plus, Periodic Acid / Schiff's, Reticulum, and Steiner special stain kits with corresponding special stain kits from two prominent manufacturers of histology stain kits. Each kit was evaluated in the following categories: packaging, procedure complexity, procedure time, microscopic result, and reagent shelf life. A rating scale of 1 (worst) to 5 (best) is used for each category; each stained slide was evaluated using the "blind method" by Don Jones, Adam Kowalczyk, and Ron Hightower. All procedures were done at room temperature or in a waterbath if specified and used the minimum procedure time listed for each step. Microwave oven procedures, if supplied, were not evaluated, as microwave ovens are not used in all laboratories and the type of microwave ovens in laboratory use vary greatly. In addition, none of the procedures were "optimized" to compensate for differences in reagent strength, dilution ratios for working solutions, or maximum versus minimum recommended procedure step times. We expect that any technician using these stain kits will have the requisite training to successfully perform the stain and sufficient experience to accurately evaluate stain results. Table 1-1: Three Companies, Four kits, rated from "1-5" in Five categories G.M.S. Kit Packaging Proc. Time Complexity St. Quality Shelf Life AMTS, Inc. 4 3 4 5 5 Comp A 3 5 3 3 4 Comp B 5 4 5 4 4 Muci Plus           AMTS, Inc. 4 5 4 4 4 Comp A 3 4 3 3 5 Comp B 5 3 5 5 5 P.A.S.           AMTS, Inc. 4 5 5 5 4 Comp A 3 5 4 3 5 Comp B 5 4 5 5 5 Reticulum           AMTS, Inc. 4 5 4 5 4 Comp A 3 5 5 4 5 Comp B * * * * * Steiner           AMTS, Inc. 4 5 4 5 4 Comp A * * * * * Comp B 5 2 5 5 5 While each Master*Tech Special Stain Kits compared very favorably to the other kits in this study, the study highlights certain areas where improvement to our kits may be achieved. One thing to keep in mind when reviewing the above data is that no "procedure tweaking" was done; an example being the different stain times used for mucicarmine staining. The minimum stain time for the working mucicarmine solution in our MUCI PLUS Stain Kit is 10 minutes, while the minimum stain time for the working solution in the other two kits is 30 minutes! In addition, no adjustments to solution strength were made; all working solutions were used as supplied in each kit. I realize that it is a common practice of histotechnicians to adjust the procedure and/or reagents supplied in commercially available special stain kits to achieve the specific requirements of their pathologist. While the format of this study does not allow exact "apples to apples" comparisons, I believe the data accurately reflect the results an experienced histotechnician could be expected to achieve if they were using any of these special stain kits for the first time. If you have any questions or comments on this study, please e-mail me by visiting "Quality Control" under the "Contact Us" page; I will be happy to respond to your e-mails. As we are on the subject of special stain kits, in this edition of the Master*Tech Magazine we are pleased to introduce our newest special stain kit, the ALCIAN BLUE - P.A.S. Stain Kit. This kit demonstrates acid mucins, neutral mucins, and glycogen. The stain procedure can be performed in 35 to 45 minutes at room temperature and contains 1 pint of 0.5% Periodic Acid, 1 pint of Schiff's Solution, 100 ml of Alcian Blue pH 2.5, and 100 ml of Modified Mayer's Hematoxylin. I hope you will give our ALCIAN BLUE - P.A.S. Stain Kit the opportunity to earn shelf space in your laboratory. AMTS: Steiner Stain Kit Micrograph. Company B: Steiner Stain Kit Micrograph. AMTS: Mucicarmine Stain Micrograph.    Company A: Mucicarmine Stain Micrograph. AMTS: PAS Stain Kit Micrograph. Company B: PAS Stain Kit Micrograph.