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?