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Electron Microscopy Sciences

Technical Data Sheets

Neat Stain™ Trichrome Stain Kit

EMS #27106-01

Summary

For the staining of stool specimens. This procedure is based upon the Wheatley trichrome technique (1951) which is a rapid staining procedure providing good results for routine examination. The method has been simplified so use of a mordant prior to staining is not necessary.

Storage Conditions

Store at controlled room temperature (15-30°C). Opened cuvettes can be resealed between uses, with the supplied adhesive backed foil strips.

Procedural Notes

The reagents may be used with individual slides or a five slide holder is available. Use 25 x 75 x 1mm slides to avoid procedural difficulties. Slides should be drained well between solutions by blotting on a paper towel two to four seconds, to minimize carry-over to the next cell. Since the acid alcohol continues to decolorize when removed from solution, compensate for the time required for it to drain when blotted and into the next cell, 95% alcohol. All reagents are necessary and included with the exception of the xylene and carbol-xylene.

Place specimen toward lower half of slide to insure it is fully immersed in reagents. Although the inner chambers of cuvettes have been designed to minimize contact with specimens, it is recommended that care be exercised to avoid scraping of slides. When resealing cuvettes with supplied foil, press foil firmly around perimeter of the cuvette. This will minimize evaporation to allow storage of an opened pack for approximately one week.

Note: Xylene maybe substituted for the carbol-xylene steps and processing time adjusted to meet user requirements.

Technique For A PVA Fixed Specimens
1. 70% alcohol/iodine solution 10 to 20 minutes
2. 70% alcohol 3 to 5 minutes
3. 70% alcohol 3 to 5 minutes
4. Trichrome Stain 8 minutes
5. 90% Alcohol, acidified 5 to 10 seconds
6. 95% Alcohol rinse briefly
7. 95% Alcohol 5 minutes
8. Carbol-xylene 5 to 10 minutes
9. Xylene 10 minutes
Technique For Unpreserved Specimens
1. Schaudinn's fixative 5 minutes @ 50°C or 1 hour at RT
2. 70% alcohol/iodine 1 minute
3. 70% alcohol 1 minute
4. 70% alcohol 1 minute
5. Trichrome Stain 2 to 8 minutes
6. Acid alcohol 5 to 10 seconds
7. 95% Alcohol rinse briefly
8. 95% alcohol rinse twice
9. Carbol-xylene 1 minute
10. Xylene 1 to 3 minutes

Mount coverslip using suitable mounting medium for both methods.

General Safety Note

Most supplied reagents are flammable liquids with the exception of Trichrome stain. Use with adequate ventilation. Avoid excessive heat, sparks and open flame. Reagents will cause eye irritation. In case of contact, flush eyes with water for 15 minutes. Wash thoroughly after handling. Contact a physician if necessary.

Limited Warranty

This product is warranted to perform as described in its labeling and product literature. Electron Microscopy Sciences disclaims all other warranties, expressed, implied and statutory with respect to this product, including, but not limited to, the warranty of fitness for a particular purpose and the warranty of merchantability. In no event shall Electron Microscopy Sciences, or any of its affiliates, be liable for incidental or consequential damages of buyer or any third party, howsoever arising, even if advised of such damages.

Product Information

Neat Stain™ Trichrome Stain Kit