PRINCIPLES OF PERIPHERAL BLOOD FILMS
Quick stains
, as the name implies, are fast and easy. Stain is purchased in a bottle as a modified Wright or Wright-Giemsa stain.
● echinocytic red blood cells ● spherocytes ● necrobiotic leukocytes ● vacuolated neutrophils
After 5 hours at room temperature causes unacceptable blood cell artifacts:
5 hours
After __ hours at room temperature causes unacceptable blood cell artifacts
4 hours
Blood films should be made within __ hours of drawing the specimen.
echinocytic
Blowing breath on a slide is counterproductive because the moisture in breath causes RBCs to become
pH dependent
Buffer: staining reactions are
-RBC orange to salmon pink -WBC nuclei should be purple to blue -Cytoplasm of neutrophils should be pink to tan with violet or lilac granules. -Eosinophils should have bright orange refractile granules.
Color of Microscopically WBC RBC Cytoplasm Eosinophils
sodium citrate tube
Corrective Action: Recollect blood using __
1. In vitro platelet satellitosis 2. Pseudoleukocytosis
EDTA SPECIMEN CONSIDERATION
film is smooth without irregularities, holes film has a "rainbow" appearance. lateral edges of the film are visible. film is finger shaped
Features of a Well-Made Wedge PBS
methylene blue azure
Giemsa stains also contain
metallic sheen (or green "scum")
In manual techniques approximately equal amount of buffer is added to the slide and a __ should appear on the slide if mixing is correct.
Manual wedge technique.
It is the most convenient and most commonly used method
3-inch x 1-inch
Manual wedge technique Requires at least two __ clean glass slides.
thiazine-eosinate complex
Oxidized methylene blue and eosin form a __ , which stains neutral components.
Use EDTA microcollection tubesp
PERIPHERAL BLOOD FILMS SPECIMEN: finger and heel puncture Corrective Action:
finger and heel puncture
PERIPHERAL BLOOD FILMS SPECIMEN
disodium or tripotassium EDTA
PERIPHERAL BLOOD FILMS SPECIMEN ( tube )
1
Quick stains whole process takes about _ minute.
Microscopically
RBCs should appear orange to salmon pink
hematology analyzer
Reanalyze using
● Pure Wright stain ● Wright-Giemsa stain
Romanowsky Stain
Manual Technique
Slides are placed on the rack, film side facing upward.
1.1
The WBC and platelet counts are multiplied by
eosin and methylene blue
These are considered polychrome stains because they contain both
Blood is nine-tenths of the total tube volume (2.7 mL of and 0.3 mL of sodium citrate).
Use the corrected dilution of anticoagulant:
Oxidized methylene blue and eosin
___ form a thiazine-eosinate complex, which stains neutral components.
Macroscopically
a well-stained blood film should be pink to purple
Neutrophils
are so named because they have cytoplasmic granules that have a neutral pH and pick up some staining characteristics from both stains.
Methylene blue
basic and stains acidic cellular components, such as ribonucleic acid (RNA).
24 hours
distilled water placed in a glass bottle for at least
Methanol
fixes the cells to the slide.
Eosin
is acidic and stains basic components, such as hemoglobin and eosinophilic granules.
pH 6.4 to 6.8
pH of distilled water placed in a glass bottle for at least 24 hours
blood films and bone marrow smears
should be dried as quickly as possible to avoid drying artifact.
Automated Slide Stainers
▪ Staining takes about 5 to 10 minutes. ▪ The processes of fixing/staining and buffering are similar in practice to those of the manual method. ▪ The slides may be automatically dipped in stain and then in buffer and a series of rinses.
