Differential Stain Kit

differential stain kit microorganism stain

(use: Hematological Staining & Special Stain for Helicobacter pylori)

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Part # 9112B500ml kit $68.50H

 

  

DIFFERENTIAL STAIN KIT FOR SMEARS & TOUCH IMPRINTS INCLUDES:

    Part 9112B
Solution A: Xanthene Stain Solution 500 ml
Solution B: Thiazine Stain Solution 500 ml
Solution C: Fixative 500 ml

     

Individual stain solutions may be available for purchase under separate part numbers.

 

Additionally Needed:

Xylene, ACS Part 1445

                                           

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

 

DIFFERENTIAL STAIN KIT APPLICATION FOR SMEARS & TOUCH IMPRINTS:

The Newcomer Supply Differential Stain Kit, a modification of the Wright Giemsa Stain technique, uses aqueous based stain solutions and a methanol fixative.  This stain kit provides a rapid 3-step process that can be used for differential assessment of:  peripheral blood smears, touch imprints, fine needle aspirations (FNA), bone marrow biopsy aspirations, as well as detecting microorganisms.                            

DIFFERENTIAL STAIN KIT METHOD FOR SMEARS & TOUCH IMPRINTS: 

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

All Newcomer Supply Stain Kits are designed to be used with Coplin jars filled to 40 ml following the staining procedure provided below.  Some solutions in the kit may contain extra volumes.                                                                 

DIFFERENTIAL STAIN KIT STAINING PROCEDURE FOR SMEARS & TOUCH IMPRINTS:

  1. Prepare within an accepted time frame, a well-made blood smear, touch imprint, FNA smear or bone marrow aspiration smear/film per your laboratories protocol, with a focus on uniform cell distribution.
  2. Allow slides to thoroughly air-dry prior to staining.
  3. Dip dried slides in Solution C: Fixative 5-10 times, one second per dip.  Allow excess fixative to drain.
  4. Dip slides in Solution A: Xanthene Stain Solution 5 times, one second per dip. Allow excess solution to drain.
  1. See Procedure Notes #1, #2 and #3
  1. Quickly rinse slides with distilled water.
  2. Dip slides in Solution B: Thiazine Stain Solution 5 times, one second per dip.  Allow excess solution to drain.
  3. Rinse slides quickly in distilled water.
  4. Allow slides to air-dry, then examine microscopically.
  5. If coverslip is preferred, allow slides to air-dry; dip dried slides in xylene and coverslip with compatible mounting medium.

 

RESULTS:

Erythrocytes: Pink to yellowish-red
Platelets: Violet or purple granules
Granulocytes  
Neutrophils: Nucleus - Dark blue to violet
  Cytoplasm - Pale pink
  Granules - Purple to lilac
Eosinophils: Nucleus - Blue
  Cytoplasm - Blue
  Granules - Red to red-orange
Basophils: Nucleus - Purple or dark blue
  Granules - Dark purple
Mononuclear Cells  
Monocytes: Nucleus - Violet
  Cytoplasm - Sky blue
Lymphocytes: Nucleus - Violet
  Cytoplasm - Dark blue
Bacteria/microorganisms: Deep blue in varying shapes
Muscle and collagen: Pale Pink
Nuclei: Blue/violet
Cytoplasm: Varying shades of light blue

    

DIFFERENTIAL STAIN KIT STAINING PROCEDURE NOTES FOR SMEARS & TOUCH IMPRINTS:

  1. The division of stains in this kit gives the user the advantage of varying dips in Solutions A and B to produce different degrees of shading and intensity.  However; never use fewer than three dips of one full second each.
  2. If more intense overall stain is desired, increase the number of dips in Solutions A and B.
  1. To increase eosinophilic staining; increase the number of dips in Solution A. 
  2. To increase basophilic staining; increase the number of dips in Solution B.
  1. If a paler stain is desired; decrease dips in Solutions A and B.
  2. If using a xylene substitute, closely follow the manufacturer’s recommendations for coverslipping application.

 

REFERENCES:

  1. Bain, B.J. “Bone Marrow Aspiration”. Journal of Clinical Pathology 54 (2001): 657-663.
  2. Cox, Charles. "Accuracy of Intraoperative Imprint Cytology for Sentinel Lymph Node Evaluation in the Treatment of Breast Carcinoma." Cancer Cytopathology 105.1 (2005): 13-20.
  3. "Guidelines of the Papanicolaou Society for Fine-Needle Aspiration Procedure and Reporting." Diagnostic Cytopathology 17 (1997): 239-247.
  4. McPherson, Richard and Matthew Pincus. Henry’s Clinical Diagnosis and Management by Laboratory Methods. 22nd ed. Philadelphia:  Elsevier Saunders, 2011. 522-535.
  5. Thompson, Samuel Wisley, and Ronald D. Hunt. Selected Histochemical and Histopathological Methods. 2nd ed. Springfield, IL: Thomas, 1966. 756-762.
  6. Modifications developed by Newcomer Supply Laboratory.  

 


 

DIFFERENTIAL STAIN KIT, HELICOBACTER PYLORI, SP. IN TISSUE SECTIONS INCLUDES:

    500ml 1 Gallon
Solution A: Xanthene Stain Solution Part 10521A Part 10521B
Solution B: Thiazine Stain Solution Part 10522A Part 10522B

   

Additionally Needed:

Helicobacter sp., Artificial Control Slides Part 4275
Xylene, ACS Part 1445
Alcohol, Ethyl Denatured, 100% Part 10841
Alcohol, Ethyl Denatured, 95% Part 10842

                                           

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

 

DIFFERENTIAL STAIN KIT, HELICOBACTER PYLORI, SP. IN TISSUE SECTIONS APPLICATION:

Newcomer Supply Differential Stain procedure, a modification of the Wright Giemsa stain technique, provides a rapid staining method for demonstration of Helicobacter pylori sp. in gastrointestinal tissue sections. Procedures for both monochromatic and polychromatic versions of the Differential Stain are provided.

 

DIFFERENTIAL STAIN KIT, HELICOBACTER PYLORI, SP. IN TISSUE SECTIONS METHOD:

Fixation: 10% Phosphate Buffered Formalin (Part 1090)

Technique:  Paraffin sections cut at 5 microns   

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

 

STAINING PROCEDURE OF THE DIFFERENTIAL STAIN KIT, HELICOBACTER PYLORI, SP. IN TISSUE SECTIONS:

  1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water. 
  1. Proceed with either the monochromatic or polychromatic staining method.

 

Monochromatic Staining Method:  See Procedure Note #1.

  1. Place slides in Solution B: Thiazine Stain Solution for 1-4 minutes depending upon staining intensity preference.
  2. Rinse slide quickly in distilled water; long enough to remove excess stain.
  3. Allow slides to air-dry in a vertical position.
  4. Dip dried slides in xylene and coverslip with compatible mounting medium.
  1. See Procedure Note #2.

 

RESULTS OF THE MONOCHROMATIC STAINING METHOD FOR THE DIFFERENTIAL STAIN KIT, HELICOBACTER PYLORI, SP. IN TISSUE SECTIONS:

Helicobacter pylori sp. Dark blue
Collagen and muscle Blue
Nuclei Blue
Cytoplasm Varying shades of light blue

 

Polychromatic Staining Method: See Procedure Note #1.

  1. Place slides in Solution A: Xanthene Stain Solution for 3-5 minutes.
  2. Drain slides briefly; go directly into Solution B: Thiazine Stain Solution for 1-4 minutes depending upon staining intensity preference.
  3. Rinse well in distilled water.
  4. Allow slides to air-dry in a vertical position.
  5. Dip dried slides in xylene and coverslip with compatible mounting medium.
  1. See Procedure Note #2.

 

RESULTS OF THE POLYCHROMATIC STAINING METHOD FOR THE DIFFERENTIAL STAIN KIT, HELICOBACTER PYLORI, SP. IN TISSUE SECTIONS:

Helicobacter pylori sp. Dark blue
Collagen and muscle Pale pink
Nuclei Blue/violet
Cytoplasm Varying shades of light blue

                                                                                                                  

PROCEDURE NOTES OF THE DIFFERENTIAL STAIN KIT, HELICOBACTER PYLORI, SP. IN TISSUE SECTIONS:

  1. The timings provided in these procedures are suggested ranges.  Optimal staining times will depend upon staining intensity preference.
  2. The elimination of dehydration steps is necessary to retain the dark stain of the organism.
  3. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and coverslipping steps.

 

REFERENCES:

  1. Potvin, Carol. “A Modified Diff-Quik Stain for Helicobacter pylori in Gastrointestinal Biopsies.” Laboratory Medicine 25.6 (1994): 389-391.
  2. Skipper, Ray, and Don DeStephano. "A Rapid Stain for Campylobacter pylori in Gastrointestinal Tissue Sections Using Diff-Quik." The Journal of Histotechnology 12.4 (1989): 303-304.
  3. Modifications developed by Newcomer Supply Laboratory.