Blood Smear Report

Blood Smear

Report

Chanaka Lakshan
8/6/2015

Table of Contents

Introduction…………………………………………………………………………………….3 Principle ………………………………………………………………………………………...3
Materials for Finger Pricks…………………………………………………………………...3

Finger-prick (capillary blood)…………………………………………………………….....3

Thin Films……………………………………………………………………………………….6
Thick Films……………………………………………………………………………………...7
Biologic causes of a poor smear………………………..................................................8
Staining smears……………………………………………………………………………….8

Preparing staining buffer ……………………………………………………………………9 Staining Reagents and Supplies…………………………………………………………....9 Staining Procedure………………………………………………………………………......10
Troubleshooting…………………………………………………………………………..….11

Reference………………………………………………………………………………………12

First prepare the buffer. The stock buffer should be kept in the refrigerator, but if not
Possible, can be stored at room temperature for several weeks. Make working buffer
Which can be stored at room temperature for a few days. Buffer should be pH 7.0 to
7.2. Although this is a higher pH than normally used to stain blood cells, the
Parasites will stain darker and be more visible under the microscope.

2. A high-quality Giemsa should be used. Not all Giemsa stains are equal in quality.
We place a layer of stain in the bottom of a glass coplin jar (about 3 mL), then add
Buffer to a level that will just cover the slides (except for frosted ends!) when they are in the jar. A little practice will tell the amount of buffer to add. Place the slides,
Back-to-back into the slots of the jar, and stain at room temperature for about 50
Minutes.

3. Remove slides, rinse by dipping a few times into plain buffer, then stand on end to dry. Some workers prefer to run a thin stream of tap water over the slide to remove all the remaining stain; we have not found this necessary. Be sure to wash out the coplin jars after each use. If not properly washed, stain builds up inside the jar and reduces the quality of

Determine if there is proper distribution of the cells on the smear.
a. Scan the edges and center of the slide to be sure there are no clumps of RBCs, WBCs or platelets. Note clumps of similar cells in the feather because they may be representative of an abnormal population. This is sometimes seen in lymphoproliferative and myeloproliferative disorders.
b. WBCs pulled to the feather end of the blood slide should NOT exceed 2-3X the number of WBCs present in the examination. Reject slides that do not fit this requirement because large cells are disproportionately dragged to the feather end which may affect differential accuracy.

High power (50x) scan
1. Find an optimal area for the detailed examination and enumerations of cells.
a) The area where approximately 50% of the RBCs show minimal overlapping and 50% are individually spaced.
b) There should be no area containing large amounts of broken cells or precipitated stain.
c) The RBCs should have central pallor.
d) Nuclei and cytoplasm of WBCs should be the proper color.
e) Platelets should be clearly visible.
Troubleshooting

1. Once the drop of blood is placed on the slide, there should be no delay in making the smear. Any delay will result in abnormal distribution of the white cells with many of these accumulating at the thin edge of the smear. Rouleaux formation and clumping of platelets may also