ENG Scientific Pneumocystis Carinii Stain Kit

Catalog No.	Quantity	Type
ES7154	2 x 250 mL	Xylene
ES7152	2 x 250 mL	Ethanol 95%
ES7150	4 x 250 mL	Modified Toluidine Blue O Stain

Catalog No. ES7154 Supplier ENG Scientific Supplier No. 7154

Description

Modified Toluidine Blue O Stain allows the Pneumocystis carinii cysts to be visualized more easily after staining. For Pneumocystis Carinii Stain Kit (Modified Toluidine Blue O). Includes: 250 mL each of Solutions I, II, III and IV.

• Pneumocystis carinii is an opportunist which causes a diffuse interstitial pneumonia in patients with impaired immune systems.
• The disease itself can be classified as epidemic or sporadic; the latter occurring in patients who have an underlying immunosuppression.
• In this procedure, a sulfation reagent of glacial acetic acid and sulfuric acid (not included) is used for the removal of background material.
• This allows the Pneumocystis carinii cysts to be visualized more easily after staining with Toluidine Blue 0.
• Diagnosis of Pneumocystis carinii pneumonia can frequently be made on bronchoalveolar lavage (BAL) or on touch preparations of pulmonary tissue (open lung biopsies and transbronchial biopsies).
• Recommended Procedures:
  • BAL Specimens:
    • Using a 50 mL tube, centrifuge for 15 minutes at 2000 x g.
    • Aspirate all but the bottom 5 mL of supernatant.
    • With a Pasteur pipette, aspirate the sediment plus approximately 1 mL of the remaining 5 mL of fluid.
    • Transfer material to a 15 mL centrifuge tube, gently mix and prepare smear by spreading a drop over a 1 cmm area.
    • If concentrated specimen is very thick or mucoid, spread over entire slide with care being taken not to make the smear too thick.
    • Dry the slides on a heating block at 50 - 55°C.
    • Allow to cool before staining.
  • Touch Preparations:
    • When dry, process in same manner as slides of BAL.
    • Preparation of Sulfation Reagent:
      • In a dry coplin jar submerged in a plastic tub filled with cool water (not below 10°C), mix 45 mL of glacial acetic acid with 15 mL of concentrated sulfuric acid and stir gently with glass rod.
    • Carefully place smears in reagent for 10 minutes.
    • Mix immediately and again after 5 minutes.
    • Gently rinse with water for 10 minutes.
    • Drain excess water and stain with Sol. I for 3 - 5 minutes.
    • Rinse with Sol. II followed by Sol. III for 10 seconds each.
    • Clear in Sol. IV, two changes each for 10 seconds and mount.
• Results: The cyst forms appear as lavender structures (cup-shaped) approximately 5 urn in diameter. The cyst outline is distinct, and the internal region stains uniformly.
• Note: A negative bronchoalveolar lavage does not rule out an infection with Pneumocystis carinii. Other diagnostic procedures may be necessary.

Specifications

• Type: Xylene
• Quantity: 2 x 250 mL
• Includes: Solution IV - Xylene
• For Use With (Application): In vitro diagnostic use

For in vitro diagnostic use.