Abstract
This article describes a procedure for performing routine three-color flow cytometric analysis for acute leukemia on lysed whole bone marrow preparations. This technique uses the combination of CD45 intensity and right-angle light scatter (RALS) to distinguish leukemic cells from normal lymphocytes, monocytes, neutrophils, eosinophils, and nucleated red blood cells. On this display, leukemic cells occupy a unique blast region characterized by intermediate CD45 density and low RALS, which, in normal marrows, contains less than 5% of the total cells. This approach was applied to 39 cases of acute leukemia and 8 cases of myelodysplasia or myeloproliferative disorders. The estimate of blasts by flow cytometric analysis was correlated highly with morphologic leukemic cell counts over a wide range. Moreover, the pattern seen on the CD45-RALS display was different for different French-American-British subtypes of leukemia, suggesting that this pattern might be useful for categorization. When CD45-peridin chlorophyll α protein was combined with other pairs of fluorescein isothiocyanate- and phycoerythrin-conjugated reagents, it was possible to set an analysis window on the leukemic blasts and display dual-parameter (ie, green vs. red fluorescence) data regarding expression of two additional markers on the leukemic population. This gating strategy was superior to traditional forward-angle versus RALS displays in that it did a better job of isolating the leukemic cells analytically.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 534-540 |
| Number of pages | 7 |
| Journal | American Journal of Clinical Pathology |
| Volume | 100 |
| Issue number | 5 |
| State | Published - 1993 |
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Keywords
- Acute leukemia
- CD45
- Flow cytometry
- Immunophenotyping
ASJC Scopus subject areas
- Pathology and Forensic Medicine
Cite this
Immunophenotyping of acute leukemia by flow cytometric analysis : Use of CD45 and right-angle light scatter to gate on leukemic blasts in three-color analysis. / Borowitz, Michael J; Guenther, K. L.; Shults, K. E.; Stelzer, G. T.
In: American Journal of Clinical Pathology, Vol. 100, No. 5, 1993, p. 534-540.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Immunophenotyping of acute leukemia by flow cytometric analysis
T2 - Use of CD45 and right-angle light scatter to gate on leukemic blasts in three-color analysis
AU - Borowitz, Michael J
AU - Guenther, K. L.
AU - Shults, K. E.
AU - Stelzer, G. T.
PY - 1993
Y1 - 1993
N2 - This article describes a procedure for performing routine three-color flow cytometric analysis for acute leukemia on lysed whole bone marrow preparations. This technique uses the combination of CD45 intensity and right-angle light scatter (RALS) to distinguish leukemic cells from normal lymphocytes, monocytes, neutrophils, eosinophils, and nucleated red blood cells. On this display, leukemic cells occupy a unique blast region characterized by intermediate CD45 density and low RALS, which, in normal marrows, contains less than 5% of the total cells. This approach was applied to 39 cases of acute leukemia and 8 cases of myelodysplasia or myeloproliferative disorders. The estimate of blasts by flow cytometric analysis was correlated highly with morphologic leukemic cell counts over a wide range. Moreover, the pattern seen on the CD45-RALS display was different for different French-American-British subtypes of leukemia, suggesting that this pattern might be useful for categorization. When CD45-peridin chlorophyll α protein was combined with other pairs of fluorescein isothiocyanate- and phycoerythrin-conjugated reagents, it was possible to set an analysis window on the leukemic blasts and display dual-parameter (ie, green vs. red fluorescence) data regarding expression of two additional markers on the leukemic population. This gating strategy was superior to traditional forward-angle versus RALS displays in that it did a better job of isolating the leukemic cells analytically.
AB - This article describes a procedure for performing routine three-color flow cytometric analysis for acute leukemia on lysed whole bone marrow preparations. This technique uses the combination of CD45 intensity and right-angle light scatter (RALS) to distinguish leukemic cells from normal lymphocytes, monocytes, neutrophils, eosinophils, and nucleated red blood cells. On this display, leukemic cells occupy a unique blast region characterized by intermediate CD45 density and low RALS, which, in normal marrows, contains less than 5% of the total cells. This approach was applied to 39 cases of acute leukemia and 8 cases of myelodysplasia or myeloproliferative disorders. The estimate of blasts by flow cytometric analysis was correlated highly with morphologic leukemic cell counts over a wide range. Moreover, the pattern seen on the CD45-RALS display was different for different French-American-British subtypes of leukemia, suggesting that this pattern might be useful for categorization. When CD45-peridin chlorophyll α protein was combined with other pairs of fluorescein isothiocyanate- and phycoerythrin-conjugated reagents, it was possible to set an analysis window on the leukemic blasts and display dual-parameter (ie, green vs. red fluorescence) data regarding expression of two additional markers on the leukemic population. This gating strategy was superior to traditional forward-angle versus RALS displays in that it did a better job of isolating the leukemic cells analytically.
KW - Acute leukemia
KW - CD45
KW - Flow cytometry
KW - Immunophenotyping
UR - http://www.scopus.com/inward/record.url?scp=0027421006&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0027421006&partnerID=8YFLogxK
M3 - Article
C2 - 8249893
AN - SCOPUS:0027421006
VL - 100
SP - 534
EP - 540
JO - American Journal of Clinical Pathology
JF - American Journal of Clinical Pathology
SN - 0002-9173
IS - 5
ER -