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FCA
Flow cytometry menjadi metode yang
lebih disukai dalam menentukan
lineage dan maturasi dari sel leukemia.
Advantage :
fast
objective
quantitative method
FCA
The scope of FCA :
Menentukan lineage dari sel leukemia
Analisa klonal, biasanya kombinasi dengan
pemeriksaan molekuler dan sitogenetika.
Analisa maturasi seluler dan heterogenitas diantara
populasi sel ganas/leukemia
Aplikasi observasi pada kontrol, monitoring terapi
dan deteksi residual minimal dari penyakit.
Penggunaan FCA
Diagnostik
Leukemia akut
Krisus blast pada CML atau transformasi leukemia pada MDS
Kronik lymphoproliferative disorder (T,B,NK)
Flow cytometer
B or T Cell
marker
B or T cell
specific Ab
B or T cell
specific Ab
B or T Cell
marker
Architecture Of A FacsCalibur
Instrument
Gedung Radioputro LT 5
Jln Kesehatan Sekip Bulaksumur
Yogyakarta telp. 0274-710 3748
R2
Pada
Pada populasi
populasi Blast
Blast ditemukan
ditemukan CD
CD
CD10
CD10
Quad Events % Gated % Total
UL
2237
34.95
22.37
UR
1
0.02
0.01
LL
4142
64.71
41.42
LR
21
0.33
0.21
KESAN :
Pada POPULASI SEL BLAST DITEMUKAN CD34 dan CD10
jogjakarta, 28 AGUSTUS 20
Laboratory doctor
Selection of antigens
A "minimal" primary panel may be used for lineage
assignment of the predominant blast population,
followed by a secondary panel of mAbs characterizing
the definite phenotype and maturational stage of the
blast population depending on the results of the primary
panel.
This sequential immunophenotyping of blasts is
associated with some savings in reagent costs but
requires more time and planning.
Selection of antigens
Lineage derivation can be precisely determined by the
analysis of functionally important cytoplasmic antigens
such as
myeloperoxidase for myeloid cells
CD79 and CD22 for B-cell lineage
CD3 for T-cell lineage
CD33 and CD13 for myeloid
CD19 for B-cell and CD7 for T-cell lineage are also
diagnostically helpful, but they are not always as precise as
the detection by cytoplasmic markers.
The additional analysis of CD65 and CD117 may increase the
sensitivity for the detection of myeloid cells.
Selection of antigens
Taken together:
myeloperoxidase plus CD13/CD33 (myeloid)
CD79 plus CD19 (B-cells)
CD3 plus CD7 (T-cells)
AML subtype
.
B-cell ALL
The further goal in B-lineage ALL is the maturational
analysis of B-cell precursor ALL subtypes, i.e. pre-pre-B
ALL (or pro-B-ALL), common B-ALL, pre-B ALL and surface
Ig-positive B-ALL.
The secondary panel should include the surface
membrane markers CD22 and CD24 as additional B-cell
associated markers, as well as CD5 for the identification
of subtypes of mature B-lymphatic neoplasias
T-ALL subtypes
The goal is the discrimination of T-ALL and mature T-cell
malignancies.
The secondary panel should include CD1a, CD2, CD5 as
well as mAbs to T-cell receptor (TcR) a/b and g/d chains.
Furthermore the determination of CD4 and CD8 and their
coexpression may be helpful in the analysis of T cell
(im)maturity