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viral immunity & pathogenesis group
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OUTLINE

Weconsidermanualcountsaslessquickbutmoreprecizethanauthomatic(electronic)sinceinthelattercasethecountsarebasedonthesizeofparticles,soeverythingthathasthesamesizeasplateletwillbecountedasanevent.Anywaybothgivecomparableresults-thrombocytopenicprobeincomparisonwithnormaloneremainsthrombocytopenicnomatterwhattypeofcountsperformed.Inourlabweuseasaroutineproceduremanualcountsandsometimesweuseautomaticcellcounterinaddition.

PROTOCOL

  1. Obtainbloodfromplexusretroorbitalisbyusingsliconizedmicrocapillaries(20ul)
  2. Immediatelydilute1:100inUnopettekits(BectonDickinson)
  3. Lettheprobestayfor1.5hrsRTforcompleteerythrocytelysis
  4. Countplateletsunderaphasecontrastmicroscopeatx400magnificationusingNeubauerhemocytometer

SOLUTIONS

    TheUnopetteReservoircontains1.98mlofdiluentmixture:Ammoniumoxalate11.45gm,Sorensen`sphosphatebuffer1.0gm,Thimerosal0.1gm,Purifiedwaterqsto1L
ADDITIONALINFO
Forautomaticcountsyoucanuse:
  • H-6000orH*2flowautoanalyzer(BayerCo)
  • electroniccellcountermodel2FChannelysermodel256(CoulterElectronics)
  • REFERENCES
    1. Bergmeier,W.,Rackebrandt,K.,Schroder,W.,ZirngIBL,H.andNieswandt,B.(2000).StructuralandfunctionalcharacterizationofthemousevonWillebrandfactorreceptorGPIb-IXwithnovelmonoclonalantibodies.Blood95(3):886-93.
    2. Bizzaro,N.andFiorin,F.(1999).CoexistenceoferythrocyteagglutinationandEDTA-dependentplateletclumpinginapatientwiththymomaandplasmocytoma.ArchPatholLabMed123(2):159-62.
    3. Teeling,J.L.,Jansen-Hendriks,T.,Kuijpers,T.W.,deHaas,M.,vandeWinkel,J.G.,Hack,C.E.andBleeker,W.K.(2001).TherapeuticefficacyofintravenousimmunoglobulinpreparationsdependsontheimmunoglobulinGdimers:studiesinexperimentalimmunethrombocytopenia.Blood98(4):1095-9.

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