Multiparametric flow cytometry of

normal human bone marrow:

analysis and display strategies

GEIL-GTLLF

2008

Part three : Monocytic maturation

III.1

Backgating

Display

CD45

SS

C

FLn

SS

C

CD45

SS

C

Strategy

FLx

FL

y

FLx

FL

y

FLx

FL

y

CD45

SS

C

CD45

SS

C

CD45

SS

C

Monober

III.2

Crossgating

Display

CD45

SS

C

FLn

SS

C

CD45

SS

C

Strategy

III.3

CD45

SS

C

CD45

SS

C

FL1

FL

2

CD45

SS

C

FL1

FL

3

FL2

FL

3

FL3+ FL2+ FL1+

Tube 1 : CD11b/CD14/CD16/CD45 Tube 5 : CD38/DR/CD34/CD45

CD11b/CD14 p. III.5 CD38/HLADR p.

III.21

CD11b/CD16 p. III.6 CD38/CD34 p. III.22

CD14/CD16 p. III.7 HLADR/CD34 p. III.23

Crossgating p. III-8 Crossgating p. III.24

Tube 2 : CD11b/CD117/CD34/CD45 Tube 7 : DR/CD117/CD34/CD45

CD11b/CD117 p. III-9 HLADR/CD117 p. III.25

CD11b/CD34 p. III-10 HLADR/CD34 p.

III.26

CD117/CD34 p. III-11 CD117/CD34 p. III.27

Crossgating p. III-12 Crossgating p. III.28

Tube 3 : CD36/CD71/CD11c/CD45 Tube 9 : CD64/CD24/CD10/CD45

CD36/CD71 p. III-13 CD64/CD24 p. III.29

CD36/CD11c p. III.14 CD64/CD10 p. III.30

CD71/CD11c p. III.15 CD24/CD10 p. III.31

Crossgating p. III.16 Crossgating p. III.32

Tube 4 : CD34/CD13/CD33/CD45

CD34/CD13 p. III.17

CD34/CD33 p. III.18

CD13/CD33 p. III.19

Crossgating p. III.20 IIII.4

Informative combinations

CD11b/CD14

Monober Gating CD11b Gating CD14

III.5

FLx

FL

y

FLx

FL

y

FLx

FL

y

CD45

SS

C

CD45

SS

C

CD45

SS

C

CD45ECDCD45ECD CD45ECD

CD11bFITCCD11bFITC CD11bFITC

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

CD

14

PE

CD

14

PE

CD

14

PE

CD11b/CD16

Gating CD11b Gating CD16Monober

III.6

FLx

FL

y

FLx

FL

y

FLx

FL

y

CD45

SS

C

CD45

SS

C

CD45

SS

C

CD45ECDCD45ECD CD45ECD

CD11bFITCCD11bFITC CD11bFITC

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

CD

16

PC

5

CD

16

PC

5

CD

16

PC

5

CD14/CD16

Monober Gating CD14 Gating CD16

III.7

FLx

FL

y

FLx

FL

y

FLx

FL

y

CD45

SS

C

CD45

SS

C

CD45

SS

C

CD45ECDCD45ECD CD45ECD

CD14PECD14PE CD14PE

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

CD

16

PC

5

CD

16

PC

5

CD

16

PC

5

Crossgating CD11b/CD14/CD16

Gating CD 16 Gating CD14 Gating CD11b

CD45

SS

C

CD45

SS

C

FL1

FL

2

CD45

SS

C

FL1

FL

3

FL2

FL

3

FL3+ FL2+ FL1+

III.8

CD45ECDCD45ECD CD45ECD

CD11bFITCCD11bFITC CD14PE

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

CD

14

PE

CD

16

PC

5

CD

16

PC

5

CD11b/CD117

III.9

Gating CD11b Gating CD117Monober

FLx

FL

y

FLx

FL

y

FLx

FL

y

CD45

SS

C

CD45

SS

C

CD45

SS

C

CD45APCCD45APC CD45APC

CD11bFITCCD11bFITC CD11bFITC

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

CD

11

7P

E

CD

11

7P

E

CD

11

7P

E

CD11b/CD34

III.10

Gating CD11b Gating CD34Monober

FLx

FL

y

FLx

FL

y

FLx

FL

y

CD45

SS

C

CD45

SS

C

CD45

SS

C

CD45APCCD45APC CD45APC

CD11bFITCCD11bFITC CD11bFITC

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

CD

34

PC

7

CD

34

PC

7

CD

34

PC

7

CD117/CD34

III.11

Gating CD117 Gating CD34Monober

FLx

FL

y

FLx

FL

y

FLx

FL

y

CD45

SS

C

CD45

SS

C

CD45

SS

C

CD45APCCD45APC CD45APC

CD117PECD117PE CD117PE

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

CD

34

PC

7

CD

34

PC

7

CD

34

PC

7

Gating CD117 Gating CD11bGating CD 34

Crossgating CD11b/CD117/CD34

III.12

CD45

SS

C

CD45

SS

C

FL1

FL

2

CD45

SS

C

FL1

FL

3

FL2

FL

3

FL3+ FL2+ FL1+

CD45APCCD45APC CD45APC

CD11bFITCCD11bFITC CD117PE

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

CD

11

7P

E

CD

34

PC

7

CD

34

PC

7

CD36/CD71

III.13

Monober Gating CD36 Gating CD71

FLx

FL

y

FLx

FL

y

FLx

FL

y

CD45

SS

C

CD45

SS

C

CD45

SS

C

CD45ECDCD45ECD CD45ECD

CD36FITCCD36FITC CD36FITC

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

CD

71

PE

CD

71

PE

CD

71

PE

CD36/CD11c

III.14

Monober Gating CD36 Gating CD11c

FLx

FL

y

FLx

FL

y

FLx

FL

y

CD45

SS

C

CD45

SS

C

CD45

SS

C

CD45ECDCD45ECD CD45ECD

CD36FITCCD36FITC CD36FITC

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

CD

11

cP

C5

CD

11

cP

C5

CD

11

cP

C5

CD71/CD11c

III.15

Monober Gating CD71 Gating CD11c

FLx

FL

y

FLx

FL

y

FLx

FL

y

CD45

SS

C

CD45

SS

C

CD45

SS

C

CD45ECDCD45ECD CD45ECD

CD71PECD71PE CD71PE

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

CD

11

cP

C5

CD

11

cP

C5

CD

11

cP

C5

Crossgating CD36/CD71/CD11c

III.16

Gating CD11c Gating CD71 Gating CD36

CD45

SS

C

CD45

SS

C

FL1

FL

2

CD45

SS

C

FL1

FL

3

FL2

FL

3

FL3+ FL2+ FL1+

CD45ECDCD45ECD CD45ECD

CD36FITCCD36FITC CD71PE

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

CD

71

PE

CD

11

cP

C5

CD

11

cP

C5

CD34/CD13

III.17

Monober Gating CD34 Gating CD13

FLx

FL

y

FLx

FL

y

FLx

FL

y

CD45

SS

C

CD45

SS

C

CD45

SS

C

CD45PC7CD45PC7 CD45PC7

CD34FITCCD34FITC CD34FITC

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

CD

13

PE

CD

13

PE

CD

13

PE

CD34/CD33

III.18

Monober Gating CD34 Gating CD33

FLx

FL

y

FLx

FL

y

FLx

FL

y

CD45

SS

C

CD45

SS

C

CD45

SS

C

CD45PC7CD45PC7 CD45PC7

CD34FITCCD34FITC CD34FITC

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

CD

33

AP

C

CD

33

AP

C

CD

33

AP

C

CD13/CD33

III.19

Monober Gating CD13 Gating CD33

FLx

FL

y

FLx

FL

y

FLx

FL

y

CD45

SS

C

CD45

SS

C

CD45

SS

C

CD45PC7CD45PC7 CD45PC7

CD13PECD13PE CD13PE

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

CD

33

AP

C

CD

33

AP

C

CD

33

AP

C

Crossgating CD34/CD13/CD33

III.20

Gating CD33 Gating CD13 Gating CD34

CD45

SS

C

CD45

SS

C

FL1

FL

2

CD45

SS

C

FL1

FL

3

FL2

FL

3

FL3+ FL2+ FL1+

CD45PC7CD45PC7 CD45PC7

CD34FITCCD34FITC CD13PE

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

CD

13

PE

CD

33

AP

C

CD

33

AP

C

CD38/HLADR

III.21

Monober Gating CD38 Gating HLADR

FLx

FL

y

FLx

FL

y

FLx

FL

y

CD45

SS

C

CD45

SS

C

CD45

SS

C

CD45ECDCD45ECD CD45ECD

CD38FITCCD38FITC CD38FITC

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

HL

AD

RP

E

HL

AD

RP

E

HL

AD

RP

E

CD38/CD34

III.22

Monober Gating CD38 Gating CD34

FLx

FL

y

FLx

FL

y

FLx

FL

y

CD45

SS

C

CD45

SS

C

CD45

SS

C

CD45ECDCD45ECD CD45ECD

CD38FITCCD38FITC CD38FITC

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

CD

34

PC

5

CD

34

PC

5

CD

34

PC

5

HLADR/CD34

III.23

Monober Gating HLADR Gating CD34

FLx

FL

y

FLx

FL

y

FLx

FL

y

CD45

SS

C

CD45

SS

C

CD45

SS

C

CD45ECDCD45ECD CD45ECD

HLADRPEHLADRPE HLADRPE

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

CD

34

PC

5

CD

34

PC

5

CD

34

PC

5

Crossgating CD38/HLADR/CD34

III.24

CD45

SS

C

CD45

SS

C

FL1

FL

2

CD45

SS

C

FL1

FL

3

FL2

FL

3

FL3+ FL2+ FL1+

Gating CD 34 Gating DR Gating CD38

CD45ECDCD45ECD CD45ECD

CD38FITCCD38FITC HLADRPE

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

HL

AD

RP

E

CD

34

PC

5

CD

34

PC

5

HLADR/CD117

III.25

Gating CD117Gating HLADRMonober

FLx

FL

y

FLx

FL

y

FLx

FL

y

CD45

SS

C

CD45

SS

C

CD45

SS

C

CD45ECDCD45ECD CD45ECD

HLADRFITCHLADRFITC HLADRFITC

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

CD

11

7P

E

CD

11

7P

E

CD

11

7P

E

HLADR/CD34

III.26

Monober Gating DR Gating CD34

FLx

FL

y

FLx

FL

y

FLx

FL

y

CD45

SS

C

CD45

SS

C

CD45

SS

C

CD45ECDCD45ECD CD45ECD

HLADRFITCHLADRFITC HLADRFITC

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

CD

34

PC

5

CD

34

PC

5

CD

34

PC

5

CD117/CD34

III.27

Monober Gating CD117 Gating CD34

FLx

FL

y

FLx

FL

y

FLx

FL

y

CD45

SS

C

CD45

SS

C

CD45

SS

C

CD45ECDCD45ECD CD45ECD

CD117PECD117PE CD117PE

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

CD

34

PC

5

CD

34

PC

5

CD

34

PC

5

Crossgating HLADR/CD117/CD34

III.28

Gating CD117Gating CD 34

CD45

SS

C

CD45

SS

C

FL1

FL

2

CD45

SS

C

FL1

FL

3

FL2

FL

3

FL3+ FL2+ FL1+

Gating HLADR

CD45ECDCD45ECD CD45ECD

HLADRFITCHLADRFITC CD117PE

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

CD

11

7P

E

CD

34

PC

5

CD

34

PC

5

CD64/CD24

III.29

Monober Gating CD64 Gating CD24

FLx

FL

y

FLx

FL

y

FLx

FL

y

CD45

SS

C

CD45

SS

C

CD45

SS

C

CD45PC7CD45PC7 CD45PC7

CD64FITCCD64FITC CD64FITC

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

CD

24

PE

CD

24

PE

CD

24

PE

CD64/CD10

III.30

Gating CD64Monober Gating CD10

FLx

FL

y

FLx

FL

y

FLx

FL

y

CD45

SS

C

CD45

SS

C

CD45

SS

C

CD45PC7CD45PC7 CD45PC7

CD64FITCCD64FITC CD64FITC

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

CD

10

PC

5

CD

10

PC

5

CD

10

PC

5

CD24/CD10

III.31

Gating CD10Monober Gating CD24

FLx

FL

y

FLx

FL

y

FLx

FL

y

CD45

SS

C

CD45

SS

C

CD45

SS

C

CD45PC7CD45PC7 CD45PC7

CD24PECD24PE CD24PE

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

CD

10

PC

5

CD

10

PC

5

CD

10

PC

5

Crossgating CD64/CD24/CD10

III.32

Gating CD24 Gating CD64Gating CD10

CD45

SS

C

CD45

SS

C

FL1

FL

2

CD45

SS

C

FL1

FL

3

FL2

FL

3

FL3+ FL2+ FL1+

CD45PC7CD45PC7 CD45PC7

CD64FITCCD64FITC CD24PE

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

SS

C-

Hei

gh

t

CD

24

PE

CD

10

PC

5

CD

10

PC

5