13. 488 nm laser + - Fluorescence Activated Cell Sorting Charged Plates Single cells sorted into test tubes FALS Sensor Fluorescence detector
14. Hydrodynamic Systems Sample in Sheath Sheath in Laser beam Piezoelectric crystal oscillator Fluorescence Sensors Scatter Sensor Core Sheath Flow Chamber ++ -- Signal direction Signal direction
15. SMALL BEAD LARGE BEAD Frequency Histogram SMALL BEAD LARGE BEAD Sample in Sheath Sheath in Laser beam Stream Charge +2KV -2KV Waste SORT RIGHT SORT LEFT SORT DECISIONS Piezoelectric crystal oscillator Last attached droplet LEFT RIGHT Sensors Sensor
32. Acute Leukemia’s Acute leukemia's ALL AML T-ALL B-ALL pre-T-ALL Common-T-ALL early T-ALL mature T-ALL pro-B-ALL C-ALL pre-B-ALL B-ALL AUL M1 M2 M3 M4 M5 M6 M7
33.
34. Model of Leukemia/Lymphoma Maturation stop Proliferation Leukemia/ Lymphoma Morphology Lymphoblast Plasma cell Immunology Pro-B-cell Plasma cell Y Y Y Y
35. B Cell Leukemia's TdT TdT 24 24 CD19 24 CD34 Pro B-ALL (BI) TdT CD22 CD79a CD19 CD10 CD22 Common B-ALL (BII) CD79a Y IgM CD19 CD20 CD22 CD79a CD24 B-ALL (BIV) CD19 CD10 CD22 CD79a Pre B-ALL (BIII)
38. AUL M0 Blast resmble L2 but –ve for lymphoid marker CD3 - /CD19 - CD33+/MPO+ blasts of a AML FAB M0
39. Biphenotypic hybrid Definition Case with typical lymphoblast or myeloblast that unexpectedly express two or more Ags of the opposit lineage on the same cells MPO CD13 CD33 CD22 CD79a CD34
40. Biphenotypic hybrid It is nessesary to score more than 2 point from 2 separate lineage Point B.Lineage markers B.lineage markers Meyloid markers 2 CD 79a Cd22 µ chaine CD 3 MPO 1 CD 19 CD 10 CD2 CD5 CD 13 CD 33 0.5 TdT TdT CD 7 CD 11 b/c CD 14/15
41. Mixed or bilinear Definition The cells show separate population of lymphoblast and meyloblast . CD 13 CD19 CD22 CD79a CD34 TdT CD33 CD34
42. Antigen Pattern I Definition Cells are clearly defined by several pan-lineage antigens and co-express an antigen of a different lineage e.g. CD65 - myeloid antigen - expressed on a immature B-cell (CD19 + , CD34 + , cyCD22 + , cyCD79a + ) CD65 CD19 CD22 CD79a CD34 TdT aberrant
43. Antigen PatternII Definition The differentiation stage of the cells are clearly defined by several antigens e.g. antigens of mature B-cells, but co-express an antigen of immature B-cells like CD34. CD34 Y IgM CD19 CD20 CD22 CD79a CD24 asynchronous
57. Diploid : Only one G0/G1 peak is observed. Aneuploid : at least 2 separate G0/G1 peaks are demonstrated. - Hyperdiploid : abnormal peak above diploid peak. - Hypodiploid : abnormal peak below diploid peak. - Multiploid : Two or more abnormal peaks. - Tetraploid : Presence of abnormal peak at G2/M position.
61. Stem Cells - Pluripotent - Reconstitution of bone marrow - 2 % in bone marrow - CD34 + - Mobilization by chemo- and factor- therapy P eripheral B lood S tem C ell T ransplantation ( PBSCT )
85. HLA-Typing Rapid and easy Detection of HLA-B27 by flow cytometry $ Clinical relevance 1. T r ansplantation 2. Proof of paternity 3. Autoimmune diseases Correlation of HLA-subtypes and incidence classifying rheumatoid diseases