Products

Trichrome Stain, Wheatley Modified

Newcomer Supply Beaker

(use: mod. of Wheatley Trichrome Stain for microsporidia, etc.)

 

Product Options:

Part # 10351A 250 ml $87.00

SOLUTION:

250 ml
Trichrome Stain, Wheatley Modified Part 10351A

 

Additionally Needed:

Alcohol, Ethyl Denatured, 70% Part 10844
Acetic Acid, Glacial, ACS Part 10010
Alcohol, Ethyl Denatured, 95% Part 10842
Alcohol, Ethyl Denatured, 100% Part 10841
Xylene, ACS Part 1445

 

For storage requirements and expiration date refer to individual bottle labels.

 

APPLICATION:

Newcomer Supply Trichrome Stain, Wheatley Modified provides a ready-to-use solution for rapid staining and permanent slide preparation for detection and identification of intestinal protozoa, flagellates and microsporidia in fecal smears.

 

METHOD:

Fixation: According to laboratory protocol for fecal/stool samples

    1. See Procedure Note #1.

Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

All Newcomer Supply stain procedures are designed to be used with Coplin jars filled to 40 ml following the provided staining procedure.

 

STAINING PROCEDURE:

    1. Prepare a well-made fecal smear from a fresh or fixed specimen with focus on uniform distribution of material.
        1. Fix fresh smears according to recommendations.
    2. Allow smears to dry for an hour at 35-37°C or overnight at room temperature.
    3. Place slides in 70% ethyl alcohol; two changes 3 minutes each.
        1. See Procedure Note #2.
    4. Stain in Trichrome Stain, Wheatley Modified for 8-10 minutes.
    5. Prepare Acid-Ethanol Solution; combine and mix well.
        1. Alcohol, Ethyl Denatured, 95% (Part 10842)         100 ml
        2. Acetic Acid, Glacial, ACS (Part 10010)                  5 ml     
    6. Differentiate slides in Acid-Ethanol Solution; 3-5 seconds.
    7. Rinse quickly in 100% ethyl alcohol; 2 dips.
    8. Dehydrate in two changes of 100% ethyl alcohol; 3 minutes each.
    9. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

PROCEDURE NOTES:

    1. Stool specimens received in modified polyvinyl alcohol (PVA) and sodium acetate-acetic acid-formalin fixatives (SAF) or freshly fixed smears in a modified Schaudinn Solution should be fixed and prepared according to manufacturer’s recommendations.
    2. Drain slides after each step to prevent solution carry over.

 

RESULTS:

Nuclear chromatin & chromatoid bodies Red to purple
Bacteria & ingested RBC’s Red to purple
Cytoplasm of cysts Blue/green with purple tinge
Cytoplasm of protozoan trophozoites Blue/green with purple tinge
Microsporidia spores Pink/red wall with clear interior
Background Green

 

REFERENCES:

    1. Bauer, John D. Clinical Laboratory Methods. 9th ed. St. Louis: Mosby, 1982. 951-952.
    2. “CDC – DPDx – Diagnostic Procedures – Stool Specimens,” cdc.gov/dpdx/diagnosticprocedures/stool/staining.html.
    3. Ryan, Norbert, G. Sutherland, K. Coughlan, M. Globan, J. Doubletree, J. Marshall, R.W. Baird, J. Pedersen, and Brian Dwyer. “A New Trichrome-Blue Stain for Detection of Microsporidial Species in Urine, Stool and Nasopharyngeal Specimens.” Journal of Clinical Microbiology2 (1993): 3264-3269.
    4. Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 250.
    5. Wheatley, W.B. “A Rapid Staining Procedure for Intestinal Amoeba and Flagellates.” American Journal of Clinical Pathology 21 (1951): 990-991.
    6. Modifications developed by Newcomer Supply Laboratory.