流式细胞术 - chem17

1 82802389 82802437 828050342 Peking University Health Science Center Peking University Health Science Center4 Peking University Health Science Center 5 Peking University Health Science Center 6 Peking University Health Science Center Flow Cytometry, FCM DNA/RNA Flow Cytometer 7 Peking University Health Science Center / Cytometric Beads Array,CBA 8 Peking University Health Science Center 1970 Phywe FCM DNA Kamensky C 1972 Len Herzenberg FACS 1973 Becton Dikinson (BD)

2 FACS-I 20 FACSA nalyzer FACScan FACSC alibur FACSLSR-II FACSV antage FACSDiva FACSAria 1975 Coulter TPS-1 EPICS C EPICS-Profile EPICS-ELITE 1994 Cytomation MoFlo 2008 BD Influx MoFlo 9 Peking University Health Science Center 10BD FACS CaliburLSRIIFACS CountFACS Canto Peking University Health Science Center11 FACS Aria IIInflux FACSV antage & DivaBD Peking University Health Science Center12 Beckman EPICS XL/XL-MCLC ytomicsTM FC 500 Peking University Health Science CenterCyAn ADP13 MoFlow XDP IIMoFlow IEPICS AltraBeckman Peking University Health Science Center14 Peking University Health Science CenterFACS CaliburFACS LSRFACS Vantage SEFACS Aria 15 Peking University Health Science Center 16 Peking University Health Science Center17 Peking University Health Science Center SSC PMT FSC 18 Peking University Health Science Center 488nm 355nm 633nm 647nm 568nm Rhodamin 6G 550-650nm BD FACS Calibur 635nm 19 2-4 488nm 633nm 355nm 407nmUV 13 Peking University

3 Health Science Center20 Peking University Health Science Center21 Peking University Health Science Center22 Peking University Health Science Center Longpass Shortpass Bandpass460 500 540460 500 540460 500 540SP 500LP 500BP500/5023 Peking University Health Science Center + 24 Peking University Health Science Center25 50 Peking University Health Science Center26 Hi Low DNA Peking University Health Science Center27 12 l/min 60 l/min Peking University Health Science Center28 Peking University Health Science Center FSC PMT.

4 SSC FL1 FL2 FL3 FL4 29 Peking University Health Science Center PMT 30 Peking University Health Science Center31 Peking University Health Science Center 32 Peking University Health Science Center33 Peking University Health Science CenterBD FACS Calibur ( 34 Peking University Health Science CenterFluorescence Activated Cell Sorting488nmlaser+-Charged PlatesSingle cells sortedinto test tubesFALS SensorFluorescence detector 35 Peking University Health Science Center36 Peking University Health Science Center 37 Peking University Health Science Center 38 FSC SSC Peking University Health Science Center39 Peking University Health Science CenterSSC.)

5 40 Peking University Health Science Center FSC 1~6 SSC 90 SSCFSC 41 SSC SSC Peking University Health Science Center42 Peking University Health Science Center43 Peking University Health Science Center DNA RNA PMT SSCFL1FL2FL3 PMT 150 ~ 999 V FL3 FL2 FL1 SSC FACS Calibur E00E01E02E03E-144 A DNA DNA W BD FL2-W G2 DNA Peking University Health Science Center ( ) 045 Peking University Health Science CenterDDM (DoubletDiscrimination Module) DNA FL2-W FL2-A BD 1 2 48 (L1) FITCA lexa Fluro 488488 PE PE-Texas RedPE-Cy5 PerCP PE-Cy7 (L2) APCA lexa Fluro 647633 APC-Cy7 (L3)

6 Alexa Fluro 405 Pacific Blue405 Peking University Health Science Center 49 2 Peking University Health Science Center530/40 BP600700585/42 BP670 LPFITCPEPC5 Laser488nm500 PerCPWavelength nmEmissionFL1FL2FL350 Peking University Health Science Center 0 FL1-%FL2 FL2-%FL154 Peking University Health Science Center FACS Vantage APC PE PE-Cy5 FITC PerCPFACS Calibur PE APC PE-Cy5 PerCP FITC S/N signal noise PE APC 55 Peking University Health Science Center pH DNA 50% 56 Peking University Health Science Center 57 Peking University Health Science Center FCM BD 58 Peking University Health Science Center FCM FCM 600 FACSAria 125 MESF CV Coefficient of variation)

7 , FCM CV 2% FCM ~ m 6000 / / 300/ 50000/ 99%59 Peking University Health Science Center FACSC alibur CaliBRITE beads FACS comp FISHPCRW estern 60 FCM a. 1 106 20%DNA G0/G1 b. c. Fc d. PBS PH e. 2h 24 Peking University Health Science Center61 / FCM PI AO 2% 84 Peking University Health Science Center62 Peking University Health Science Center DNA RNA / / Ca2+ Mg2+ pH ROS 63 Peking University Health Science Center CD34+ CD44+ CD24+ CD133+ FISH PCR

8 Western 64 Peking University Health Science Center FCM BD 65 Peking University Health Science Center FCM 1. 2. 1 3. 66 Peking University Health Science Center FCM Histogram (Dot Plot) (Contour Plot) (Density Plot) (Pseudo 3D Plot) (3D Plot)67 Peking University Health Science Center Histogram68 Peking University Health Science Center Dot Plot69 Peking University Health Science Center Contour Plot Density Plot70 Peking University Health Science Center Pseudo 3D Plot 3D Plot71 Peking University Health Science Center FCM BD 72 Peking University Health Science CenterBD CellQuest: FACSComp.

9 1 SimulSET/MultiSET 2 ProCOUNT / CD34+ 3 HLA-B27 T HLA-B27 4 ReticCOUNT 1 ModFit DNA 2 QuantiCALC ABC 3 PAINT-A-GATE 4 ATTRACTOR FACS Diva73 Peking University Health Science CenterCellQUEST 1 FACSComp FACSComp 10000 - - Gate( ) Region( )- Marker( )- Quadrant Stats - 74 Peking University Health Science CenterModFit DNA 2 DNA G2MG1s01000 G2M200400600800G0G1sDNA 2N4N75 Peking University Health Science CenterDNA CV DNA G0/G1 CV FCM CV 2% CV= / 100% DNA DNA index, DI) G0/G1 2 G0/G1 Ploidy DNA DI=1 2c DI=2 CV 5% S SPF S PI 76 Peking University Health Science CenterDNA CV CV DNA CV CV CV 3 5% G1 CV 8 S DNA G2/M G0/G1 2 G2/M G0/G1 S 2 DDM Doublet Discrimination Module)

10 77 Peking University Health Science CenterDNA DNA QC Particles) A CEN PMT 4 1 2 3 4 B CTN DDM C 2 m CV CEN CTN CV D PI78 Peking University Health Science Center DDM (Doublet Discrimination Module) FL2-W FL2-A 1 2 3 1 2 79 Peking University Health Science CenterDDM 3 R180 Peking University Health Science CenterDNA CellQuest ModFit FACSComp FL2-W FL2-A FL2-A FL2-W DNA QC 3 2 +2 2 ModFit 81 Peking University Health Science CenterDNA B~D A 2 DebrisA ggregatesDip G1 Dip G2 Dip SDebrisA ggregatesDip G1 Dip G2 Dip SA n1 G1A n1 G2A n1 SDebrisA ggregatesDip G1 Dip G2 Dip SA n1 G1A n1 G2A n1 SABACD82 Peking University Health Science Center PI -FluorescenceEvents G0/G1 83 Peking University Health Science Center