HEMA 2 LAB - PRELIMS - BLOOD SMEAR
• platelet clumping: (direct drop or heparinized) • limited amount of blood can be made
Capillary blood Disadvantage
Eosin
▪ Acid dye ▪ Stains basic (eosinophilic) ▪ Stains hemoglobin and eosinophilic granules
Methylene Blue
▪ Basic Dye ▪ Stains acidic (basophilic) ▪ Stains Nuclei and ribonucleic acid
E. spread F. Dirt or grease, elevated lipids G. Uneven H. dry
E. Drop of blood not allowed to ___ across the width of slide F. ___ on the slide; may also due to ___ in the blood specimen G. ___ pressure on the spreader slide H. Time delay; drop of blood began to ___
Blood Artifacts RBC (Spherocytes, necrobiotic leukocytes and vacuolated neutrophils) Platelets satellitosis
EDTA Tube Disadvantages
PANOPTIC
Ex. Jenner + May-Grunwald + Giemsa. To better observe the cytoplasm
Pseuothrombocytopenia
False decrease of platelet count
The film is __ to the __ of the length of the slide
Features of a well-made wedge peripheral blood film #1: 2/3 to the ¾
The film is ___ shaped
Features of a well-made wedge peripheral blood film #2: finger
The ___ edges of the film is visible
Features of a well-made wedge peripheral blood film #3: lateral
The film is smooth without ___
Features of a well-made wedge peripheral blood film #4: irregularities, holes, or streak
When the slide is held up to the ___, the thin portion of the film has a ___ appearance.
Features of a well-made wedge peripheral blood film #5: light, "rainbow"
The __ is picked up and spread.
Features of a well-made wedge peripheral blood film #6: whole blood
Monocytes
Gray ground-glass cytoplasm w/ many tiny red purple granules
BLOOD SMEAR
Important in CBC and View cellular components
o Film Slide (3' x 1', 75 x 25 mm) o Spreader o Syringe o Lancet - for capillary blood sample o EDTA tube - for venous blood
MANUAL WEDGE TECHNIQUE (MATERIALS NEEDED)
○ Too large drop of blood ○ Too fast ○ Too high angle of spreader
MANUAL WEDGE TECHNIQUE: THICKNESS
○ Too thin drop of blood ○ Too slow ○ Too low angle of the spreader
MANUAL WEDGE TECHNIQUE: THINNESS
BLOOD SMEAR: Staining
Make the cells more visible and to allow their morphology to be evaluated
• Venous Blood • Fresh Blood
OTHER SOURCE OF SPECIMEN
Eosinophils
Orange granules
Neutrophils
Pinkish-tan cytoplasm
EDTA Tube
Prevents platelets from clumping on the glass slide. Can be stored within 4 hours. Can still produce adequate blood smears
WBC nuclei
Purple-blue to lilac
Salmon pink
RBCs
- Wright or Wright Giemsa - Leishman - May-Grundwald - Jenner
ROMANOWSKY STAIN Modifications:
COVERSLIP TECHNIQUE OR EHRLICH
Rare method for peripheral blood films
cytoplasm
Red-purple granules
Flood the slide completely. Stain should remain on the slide at least __ to fix the cells to the glass
STAINING OF PERIPHERAL BLOOD FILMS STEP #1; 1 to 3 minutes
Then an approximately equal amount of buffer is added to the slide. -A ___ (or green "scum") should appear on the slide if mixing is correct.
STAINING OF PERIPHERAL BLOOD FILMS STEP #2; metallic sheen
The mixture is allowed to remain on the slide for ___ (bone marrow smears take longer to stain than peripheral blood films).
STAINING OF PERIPHERAL BLOOD FILMS STEP #3; 3 minutes or more
MANUAL WEDGE TECHNIQUE
Should be dried immediately to avoid drying artifacts (water droplets).
Reddish-brown; Rodak: reddish to purple)
Smear
SUPRAVITAL STAIN
Stain acidic cellular components. NEW METHYLENE BLUE OR BRILLIANT CRESYL BLUE
MANUAL WEDGE TECHNIQUE COVERSLIP TECHNIQUE OR EHRLICH
TECHNIQUES OF BLOOD SMEAR
SUPRAVITAL STAIN
The Cytoplasmic RNA is precipitated into a reticulum-like network ("reticulo"-cyte). Acidophilic in Nature
COVERSLIP TECHNIQUE OR EHRLICH
Used for bone marrow aspirates smears
ROMANOWSKY STAIN
Used for: - Peripheral blood smears & bone marrow smears - Differential WBC count - Study of RBC morphology
Too thin drop of blood
Will result to a short and thin blood smear which is not acceptable because the smear should be ⅔ long
QUICK STAINS
are fast and easy. The whole process takes about 1 minute. Stain is purchased in a bottle as a modified wright or Wright + Giemsa stain.
Capillary blood
best specimen
PANOPTIC
"Pappenheim's Stain" Improves the staining of cytoplasmic granules and other bodies.
Too fast
= low pressure applied by the hands = hence, the blood will not be spread properly. = thick smear
A. Chipped or rough B. Hesitation C. too quickly D. too small
A. ___ edge on spreader slide B. ___ in forward motion of spreader slide C. Spreader slide pushed __ D. Drop of blood ___
COVERSLIP TECHNIQUE OR EHRLICH
Advantages: Excellent leukocyte distribution
Blood Artifacts
if smear is stored in RT for 5 hours
BECKMAN COULTER
An automated slide making a staining system
MANUAL WEDGE TECHNIQUE
Angle: 30-45 degrees
MANUAL WEDGE TECHNIQUE
Avoid blowing on slides to prevent moisture accumulation which leads to echinocytic (crenated) red blood cells
▪ Numbers - for RBC/WBC counting ▪ Size ▪ Maturity - an RBC is mature if it does not have a nucleus, biconcave, and is smaller ▪ Morphology - an RBC is biconcave ▪ Structure - it is important to know the structure of RBC to know if it has abnormalities
Blood Smear evaluates?