Detailed manual Red Blood Cell Count (RBCs count) – The Method Only

Detailed Manual Red blood cell count-The method only

Here we will talk about the method only and we don’t mention the condition in which the RBCs increased or decreased but we will mention it in anther posts.

Tools:

1) Counting chamber (hemocytometer)

The counting chamber recommended for cell counting is

–       A metallized surface (‘Bright-line’)

–       Double cell Improved Neubauer ruled chamber.

Non-metallized haemocytometers: While these are less expensive, they are not recommended because the background rulings and cells are not as easily seen which makes it is more difficult to count WBCs reliably using this type of chamber . Non-metallized chambers are also more difficult to fill.

Improved-Neubauer-ruled-counting-chamber-non-metalized2

Improved-Neubauer-ruled-counting-chamber-metalized3



cover glass for hemocytometer

2) Counting chamber cover glasses

Special optically plane cover glasses of defined thickness (designed for use with hemocytometers) are required.

Other cover glasses must not be used.

Manufacturers of counting chambers provide two cover glasses with each chamber.

The laboratory should always keep in stock spare cover glasses.

220px-Pasteur_PipetsPasteur-Pipette3) Pipettes/calibrated capillaries and safe filling device

We need 3 types of Pipettes

First: Graduated Pipette of 1 ml volume to get the volume needed from the diluting fluid.

Second: Pasteur pipette of a capillary pipette to mix the blood sample and with diluting fluid

Third: Calibrated pipette of 20 µl (0.02 ml, 20 cmm) to aspirate 20 µl accurately

 Sahli PipetteIn case of Calibrated Pipette:

==> You may buy a Calibrated pipette of 20 µl (0.02 ml, 20 cmm) e.g. Sahli is required to measure blood samples.

==> Or the calibrated pipette may be supplied with hemocytometer by some manufacturers and it is called a Bulb Pipette. And there is 2 Bulb pipettes supplied; one for Red blood cell count and one for WBCs count,>>>>>>  

BUT:

  • Bulb Pipettes should not be used for counting WBCs because it is not possible to obtain reliable mixing of the blood and diluting fluid inside the bulb of the pipette.

  • Bulb pipettes are also difficult to clean and expensive to replace

A safe pipette/capillary filler should be used to aspirate and dispense the blood. This can be a simple bulb filler or device.  (CLIKC HERE TO SEE THE Pipette filter)

blood-diluting-pipettes-for-white-corpuscles

blood-diluting-pipettes-for-red-corpuscles

They are differ in the volume

You can use Variable Micropipette instead of all the THREE types

micropipettes-a-volume-variable

4) Hand counter

hand-tally-counterTo count white cells accurately, a simple inexpensive mechanical hand tally counter or bead counter

Turning the knob on the side returns the counter to zero after each count.

Bead-Score-Counter

See the reagents and procedure in page 2

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