flow analysis by XWNj40k

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									Oxidative Burst
    DCFH-DA


              DCFH-DA


              DCFH      H 2O 2



              DCF
                     Fluorescent Dyes

                                 Wavelength
                      400 nm      500 nm      600 nm   700 nm

               Excitation
               Emission
Rla eIn nity
 e tiv te s




                            Fluorescein (FITC)
                         Fluorescent Dyes

                                 Wavelength
                       400 nm     500 nm      600 nm   700 nm
                Excitation
                Emission
Rla eIn n ity
 e tiv te s




                             Phycoerythrin (PE)
            Immunophenotyping


                                    CD2
            CD4

CD # = cluster designation number
                     Immunophenotyping
   5
   1




            0
  1
  1




            0
        7
Count


            0
        3



            0
            0




                .1      1     10       100   1000

                            Log FITC
           CD4/CD8 Quadstats
                           1                             2
100


                     45%                        2%
      10
      1




                     27% 3                      26%      4
           .1




                .1         1     10       100         1000
                      Log FITC Fluorescence (CD8)
         Normal Cell Cycle
     M                G0
G2                                DNA Analysis
         G1
                                  G0G1
s
              C
              o
              u
              n
              t
                                        s         G2 M
                  0        200    400       600    800   1000
                                 2N          4N
                                   DNA content
         300
         225
         150                  DNA Analysis
Counts

           75
                0




                    0   200         400   600        800   1000

                               2N               4N
                                PI Fluorescence
         300                  DNA Analysis

                                         DNA index 1.21
         225




                                      Aneuploid peak
         150
Counts

           75
                0




                    0   200     400     600       800   1000

                                PI Fluorescence
Chromosome Analysis
 (Bivariate Flow Karyotyping)
            chromosome 1


      chromosome 2
                      Reticulocyte Analysis




                                               15 0
 150




                                               112
 112




                          RMI = 34                                         RMI = 0



                                               C7t5
                                               on
Cut7 5




                                                u
 on




                      R4 R3 R2 R1                                   R4 R3 R2 R1
   37




                                                  37

                                                       0
         0




             .1   1     10    100       1000               .1   1     10      100    1000
                  log Thiazole Orange                           log Thiazole Orange
                     Calcium Flux - Indo-1


                        1000
                        800
RATIO [short/long]




                        600
                        400
                        200




                                                 Stimulation
                               0




                                   0   36   72    108        144   180
                                            Time (Seconds)
                         Light Scatter Gating
                                               Side Scatter Projection




                          1000
                                               Neutrophils




                                                                             Forward Scatter Projection
Scale
                          800
Forward Scatter




                  1000
                  200
                  100
                          600




                  50
                  40
                  30                                     Monocytes
                          400




                  20
                  15
                                                   Lymphocytes
                          200




                  8
                                 0




                                     0   200       400    600   800   1000

                                                  90 Degree Scatter
         Phagocytic Vacuole
NADPH Oxidase

NADPH
           O2
NADP          O 2-

        SOD
                              
                 H2O2


                        OH-

								
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