BRILLIANT CRESYL BLUE FOR RETICULOCYTES

IVD In vitro diagnostic procedure

St abi l i t y: 2 years under r ecommended s t orage condi t i ons.

Storage conditions: Store at 15 - 25 °С, protected from fire and sunlight. Packing to be stored tightly closed.

Procedure duration: ~35 min.

Content: Reagents:
Brilliant Cresyl Blue physiological solution

Application:

Blood smears examination for reticulocytes.

Br i l l iant cresyl blue i s more used i n hemat ol ogy l abor at or i es than i n hi st ol ogy l
a bor a t or i e s. I t i s i nval uabl e for t he s t aining of ret i cul ocyt es whereas i t s use in hist ol ogy i
s l i mi ted t o vi t al s t aining.
Reticulocytes are juvenile red cells, which contain remnants of basophilic ribonucleoproteins.
This material reacts with brilliant cresyl blue to form a bluish black precipitate of granules or filaments.
The most immature reticulocytes have the largest amount of precitable material. In the least immature
only a few dots or short strands are seen.
As the number of reticulocytes in the peripheral blood is a measure of erythropoietic activity, a
reticulocyte count is one of the essential procedures of diagnostic haematology.

N. B!: The clinical i nterpretation of any s t a ining, or i t s absence, should be complemented by mo rphological
s t udies and proper controls, and should be evaluated within t he cont ext of t he patient’ s c l i nical history and
other diagnostic t ests by a qualified pathologist.
All r eagents have been optimally prepared for use and r equire no mixing or diluting.

Method:

1. Mix equal volumes of reagent A and fresh blood (without EDTA) and pull out in
haematocrit capillare.
2. Leave the mixture in the haematocrit capillare for 30 min at room temperature.
3. Make thin smears from mixture drops on clear slides.
4. Let smears dry on air.
5. Examine without fixation or contra-staining!
Results

Reticulocytes - greenish to blue with dark blue ribonucleoprotein granules inside

Erythrocytes – greenish to blue

References:
1. Dacie J. and Lewis S., Practical haematology. Churchill Livingstone, London, 1996
2. Horobin, R W, and Kiernan, J. Conn's biological stains, Ed 10. (2002), BIOS Scientific Publishers, Oxford,
UK.
3. Edward Gurr, (1960), Encyclopaedia of microscopic stains, Leonard Hill (Books), London, England.
4. Green, F J, (1990), The Sigma-Aldrich handbook of stains, dyes and indicators Aldrich Chemical
Company, Inc., Milwaukee, Wisconsin.
5. DAKO

Producer: Emmonya Biotech Ltd, Chelopech, Sofia area, Bulgaria