viral immunity & pathogenesis group
来自 : 蚂蚁淘
OUTLINEWeconsidermanualcountsaslessquickbutmoreprecizethanauthomatic(electronic)sinceinthelattercasethecountsarebasedonthesizeofparticles,soeverythingthathasthesamesizeasplateletwillbecountedasanevent.Anywaybothgivecomparableresults-thrombocytopenicprobeincomparisonwithnormaloneremainsthrombocytopenicnomatterwhattypeofcountsperformed.Inourlabweuseasaroutineproceduremanualcountsandsometimesweuseautomaticcellcounterinaddition.
PROTOCOL
- Obtainbloodfromplexusretroorbitalisbyusingsliconizedmicrocapillaries(20ul)
- Immediatelydilute1:100inUnopettekits(BectonDickinson)
- Lettheprobestayfor1.5hrsRTforcompleteerythrocytelysis
- Countplateletsunderaphasecontrastmicroscopeatx400magnificationusingNeubauerhemocytometer
SOLUTIONS
TheUnopetteReservoircontains1.98mlofdiluentmixture:Ammoniumoxalate11.45gm,Sorensen`sphosphatebuffer1.0gm,Thimerosal0.1gm,Purifiedwaterqsto1L
ADDITIONALINFOForautomaticcountsyoucanuse:H-6000orH*2flowautoanalyzer(BayerCo)electroniccellcountermodel2FChannelysermodel256(CoulterElectronics)
REFERENCES - Bergmeier,W.,Rackebrandt,K.,Schroder,W.,ZirngIBL,H.andNieswandt,B.(2000).StructuralandfunctionalcharacterizationofthemousevonWillebrandfactorreceptorGPIb-IXwithnovelmonoclonalantibodies.Blood95(3):886-93.
- Bizzaro,N.andFiorin,F.(1999).CoexistenceoferythrocyteagglutinationandEDTA-dependentplateletclumpinginapatientwiththymomaandplasmocytoma.ArchPatholLabMed123(2):159-62.
- 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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