[PROTOCOLS>PLATELET-RELATED>PLATELET COUNT]

OUTLINE
PROTOCOL
SOLUTIONS
ADDITIONAL INFO
REFERENCES

---

OUTLINE

We consider manual counts as less quick but more precize than authomatic (electronic) since in the latter case the counts are based on the size of particles, so everything that has the same size as platelet will be counted as an event. Anyway both give logical results- thrombocytopenic probe in comparison with normal one remains thrombocytopenic no matter what type of counts performed. In our lab we use as a routine procedure manual counts and sometimes we use automatic cell counter in addition.

PROTOCOL

1. Obtain blood from plexus retroorbitalis by using sliconized microcapillaries (20ul)
2. Immediately dilute 1:100 in Unopette kits (Becton Dickinson)
3. Let the probe stay for 1.5 hrs RT for complete erythrocyte lysis
4. Count platelets under a phase contrast microscope at x400 magnification using Neubauer hemocytometer

SOLUTIONS

The Unopette Reservoir contains 1.98ml of diluent mixture: Ammonium oxalate 11.45gm, Sorensen's phosphate buffer 1.0gm , Thimerosal 0.1gm, Purified water qs to 1L

ADDITIONAL INFO

For automatic counts you can use :

H-6000 or H*2 flow autoanalyzer (Bayer Co)

electronic cell counter model 2F Channelyser model 256 (Coulter Electronics)

REFERENCES

1. Structural and functional characterization of the mouse von Willebrand factor receptor GPIb-IX with novel monoclonal antibodies. Wolfgang Bergmeier, Kirsten Rackebrandt, Werner Schröder, Hubert Zirngibl, and Bernhard Nieswandt. Blood 2000 95: 886-893
2. Coexistence of Erythrocyte Agglutination and EDTA-Dependent Platelet Clumping in a Patient With Thymoma and Plasmocytoma. Nicola Bizzaro,Franceso Fiorin; Archives of Pathology and Laboratory Medicine: Vol.123 No2 p159-162
3. Therapeutic efficacy of intravenous immunoglobulin preparations depends on the immunoglobulin G dimers: studies in experimental immune thrombocytopenia; Jessica L. Teeling et al. Blood 2001 98: 1095-1099
   

created by Andrei Musaji  updated : 2003