visual platelet count experiment

Summary

Knowledge of platelet counting principles and techniques

Operation method

visual platelet count experiment

Principle

Urea solution lyses red blood cells and white blood cells while preserving platelets in their intact form, which are diluted and counted directly in the blood cell counting place to find the number of platelets per liter of blood volume.

Materials and Instruments

Platelet Diluent Urea

Move

I. Experimental reagents: platelet diluent urea (uren) 10g / sodium citrate 0.5g / 40% formaldehyde 0.1ml / distilled water added to 100mL completely dissolved over iha , stored in the refrigerator . IV. Experimental methods:

l, clean small test tube add platelet diluent 0.38ml.

2, first let the finger warm filled straight, skin disinfection to dry after a deep stab so that the blood flow naturally wipe off the small amount of blood just appeared, (the best use of venous anticoagulant blood).

3, with a micropipette quickly and accurately suck the first natural flow of 20 microliters of blood, blood to avoid bubbles or many times up and down sucking, take a good and then immediately gently inject the blood into the bottom of the test tube of the diluent, suck the supernatant rinsing pipette three times, and gently mixed.

4, wipe and put the counting plate and coverslip, the test tube will be gently shaken for 1 minute, use the pipette to suck a drop of suspension, injected into the counting pool, to avoid bubbles or overflow, and leave it for 10-15 minutes to wait for the platelets to sink.

5、First, use low magnification to find the central large square, change high magnification to count the platelets in the central large square, the platelet difference in each middle frame shall not exceed 10, such as the number of platelets in each middle frame is less than 1, you should count the whole frame or another drop of counting plate, and then count a large frame as a control.

6、Calculation: platelets obtained in the 5 middle ^{squares×109/L}

Requirements for reporting platelet counts are ^{ΔΔΔ×109/L}.

Caveat

1, all utensils and diluent to be particularly clean, microscope carefully wiped clean, without any spot glass instruments clean and smooth, because the microscope eyepieces, objectives, counting pool and the cover piece of the dust on the platelet can be mistaken for platelets.

2， stabbing blood must be deep (3mm).

3、Platelets are easy to focus, before dropping into the counting cell to mix thoroughly, too much force will destroy the platelets, too small will not be evenly distributed, so we must grasp the appropriate.

4， must wait for the platelets to sink completely before counting, counting fast and carefully, do not regard the contamination particles as platelets. Platelet structure and leukocyte cytoplasm is similar, in the light and focus adjusted appropriately translucent with a slight refractive, edge and center of the structure of the same, with a certain form. The difference in size is generally not very large, while the size of the contaminated particles vary greatly, without a certain form, when rotating the regulator, sometimes very dark, sometimes very bright. Reference value: 1OO-300×109/L

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