Techniques for blood film staining for Differential White Blood Cell Count – Manually

أقرأ الترجمة بالبلدى

Techniques for blood film staining for Differential White Cell Count Manually

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blood film staining

The Index

Introduction ——————————————————————————- Page 1

1) Value of the test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 1

2) The principle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 1

Preparation of Thin Blood Film ——————————————————— Page 2

1) Materials and Reagents. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 2

2) Specimen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 2

3) Method for Preparations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 3

4) Drying of the film . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .Page 4

5) Writing the Name and number of the patient: . . . . . . . . . . . . . . . . . . . . . . . . Page 4

6) Fixation of the film . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  Page 4

Staining of The Blood Film ————————————————————— Page 5

Method One: Leishman staining technique . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 5

Method Two: Writ’s stain technique . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .Page 6

Method Three: May-Grunwald and Giemsa stains technique . . . . . . . . . . . .Page 7

Method Four: MField stain Technique ….. (Rapid) . . . . . . . . . . . . . . . . . . . . . Page 8

How to remedy poor results ————————————————————— Page 9

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Introduction

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Deferential White Cell Count performed through 4 main stages:

  1. Preparation of thin blood film
  2. Staining of the thin film
  3. Microscoping examination
  4. Reporting the abnormal findings

Actually in this test we not only Count the number of each type of the WBCs but also examine the morphology of RBCs and WBCs and check the presence of any parasites

Therefore we can name the test as Preparation and examination of Blood film tests not only Deferential white cell count.

But in this test we will show the staining techniques of the blood films but the microscope examination will be shown in the Complete blood count (CBC) ((((coming soon))))


1) Value of the test

Examination of thin blood films is important in the investigation and management of anaemia, infections, and other conditions which produce changes in the appearance of blood cells and differential white cell count.

A blood film report can provide rapidly and at low cost, useful information about a patient’s condition. 

Important: Reliable blood film reporting is only possible when laboratory staff are trained adequately in the recognition of blood cells and follow standardized procedures for preparing and staining blood films, reporting morphological changes and performing a differential white cell count.

2) Principle:

A thin blood film is prepared by spreading a small drop of blood evenly on a slide so that there is only one layer of cells.

Thin blood films are stained with Romanowsky stains, which contain essential azure B and eosin dyes .

The Romanowsky stains most widely used include:

  • Leishman stain, which is used alone.
  • May–Grünwald stain, which is used with Giemsa stain.
  • Giemsa stain, which can be used alone or together with May–Grünwald or Jenner stain.
  • Field stains A and B, which are prepared in water unlike the above-mentioned stains, which are made up in methanol. Field stains are used for both thin and thick blood films.

The Romanowsky stains prepared in methanol can be used to fix thin films before being diluted on the slides to stain the films. Better results are obtained by fixing first with methanol, then staining with pre-prepared diluted stains, as described below.

After staining, blood films are used for:

  • — determining leukocyte type number fractions
  • — detecting abnormal erythrocytes
  • — identifying certain parasites
  • — estimating the number of thrombocytes

Preparation of Thin Blood Film ===> Page 2

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