1 Flow Cell Injector Tip Fluorescence signals Focused laser beam Sheath fluid Purdue University Cytometry Laboratories

2 Forward Angle Light Scatter FALS Sensor Laser Purdue University Cytometry Laboratories

3 90 Degree Light Scatter FALS Sensor 90LS Sensor Laser Purdue University Cytometry Laboratories

4 Antygeny najbardziej charakterystyczne dla : limfocytów T limfocytów B komórek NK monocytów granulocytów CD2 CD19 CD16 CD14 CD13 CD3 CD20 CD56 CD4 CD33 CD4 CD21 CD8 HLA-DR CD15 CD5 CD22 HLA-DR CD7 CD23 CD8 HLA-DR sIg(G,D,M,E,A) Antygen wspólny dla całej populacji leukocytów – CD45

5 Leukaemia panel G1/G1 G1/G1 CD14/CD45 CD14/CD45 CD2/CD19 CD2/CD19 CD3/CD22 CD3/CD22 CD10/CD19 CD10/CD19 CD5/CD20 CD5/CD20 CD4/CD8 CD4/CD8 HLA-DR/CD34 HLA-DR/CD34 CD7/CD13 CD7/CD13 CD7/CD33 CD7/CD33 CD15 CD15 cyt µ (IgM) surface immunoglobulins (if CD20+) IgM IgG IgA Kappa/lambda

6 Antygeny dominujące w różnych typach białaczki: Ostra białaczka mieloblastyczna – AML CD45, CD13, CD33, CD34, CD15, HLA-DR, cyt MPO Ostra białaczka limfoblastyczna z linii B – ALL B CD45, CD19, CD22, CD20, HLA-DR, CD34, cyt. łańcuch , Ig powierzchniowe cyt.CD22 Ostra białaczka limfoblastyczna z linii T – ALL T CD45, CD2, CD3, CD4, CD5, CD8, CD34, cyt.CD3

7

8 ALL-T

9 IgG/IgG

10 CD2/CD19

11 CD3/CD22

12 CD4/CD8

13 CD5/CD20

14 CD10/CD19

15 HLA-DR/CD34

16 CD7/CD13

17 cyt CD3

18 ALL-B Common ALL B coexpression of myeloid antigens

19 CD2/CD19

20 CD3/CD22

21 CD5/CD20

22 CD10/CD19

23 CD4/CD8

24 HLA-DR/CD34

25 CD15

26 CD7/CD13

27 CD7/CD33

28 CD19/CD33

29 CD19/CD13

30 ALL-B ALL – proB ALL – proB CD34 CD34 HLA-DR HLA-DR CD19 CD19 CD22 CD22 cyt cyt ALL common ALL common CD34 CD34 HLA-DR HLA-DR CD10 CD10 CD19 CD19 CD20 (var.) CD20 (var.) CD22 (var.) CD22 (var.) ALL – preB HLA-DR CD19, CD22 heavy chain µ (cyt IgM) ALL – B mature cells HLA-DR CD19 CD20 CD22 heavy chain µ (cyt IgM) 1 of light kappa or lambda chain