TheStainingprocedureiscarriedoutat4^oCanddark.Appropriatecontrolsareneededtogetcorrectresults,suchasunstainedcontrol,isotypeantibodystainedcontrolandpositivecontroletc.

1. MakesinglecellsUSPensioninL-15ataconcentrationof1-2x10⁷cells/ml
2. Add50ulcellsuspensionto5mlpolystyrenetube(or96wellV-bottompalte)
3. Add50ulofappropriatelydilutedlabeledantibodytothecellsandmixgentlely
4. Incubateat4^oCindarkfor30minutes
5. Washcellsbyadding2mlL-15andcentrifuge5minutesat1000rpm(300xg)For96wellplate,washcellsthreetimeswith150ulL-15
6. Resuspendstainedcellpelletsin400ulL-15at4^oCforflowcytomery
7. Cellscanberesuspendedinfixationsolutionandstoredfor1week(optional)
8. Add10ulpropidiumiodidepriortoanalysistodetectdeadcells(optional)
9. CalibratetheFACScanbystandardfluorescentbeads(optional)

Flowcytometersarecomplexinstrumentsthatrequireawell-trainedoperator.PriortousetheBectonDickinsonFACScan,takeatrainingcoursewithanexperienceduser.Followtherulespostedintheworkingareaforappropriatemaintenanceoftheinstrument.

1. TurnontheFACScanpowerandwaituntiltheindicatorlightchangesfromNOTREADYtoSTANDBY.RestartthecomputertoconnectwiththeFACScan.
2. Openyouraccountonthedesktopandusetheappropriatetemplateforaquiringdata(seereferenceformakingcorrectsetting).
3. TurnfluidcontrolvalvetoRUNandplacesampletubeonFACScan
4. AnalysisdatawithCELLQuestSoftware

References

1. BectonDickinsonImmunocytometrySystems
2. CurrentProtocolsinImmunology

================  蚂蚁淘在线  ================

免责声明：本文仅代表作者个人观点，与本网无关。其创作性以及文中陈述文字和内容未经本站证实，对本文以及其中全部或者部分内容、文字的真实性、完整性、及时性本站不做任何保证或承诺，请读者仅作参考，并请自行核实相关内容

版权声明：未经蚂蚁淘在线授权不得转载、摘编或利用其他方式使用上述作品。已经经本网授权使用作品的，应该授权范围内使用，并注明“来源：蚂蚁淘在线”。违反上述声明者，本网将追究其相关法律责任。