scholarly journals Flow cytometric analysis of cellular endogenous fluorescence simultaneously with emission from exogenous fluorochromes, light scatter and absorption

Cytometry ◽  
1981 ◽  
Vol 2 (1) ◽  
pp. 39-43 ◽  
Author(s):  
Bo Thorell
Keyword(s):  
Flow Cytometric Analysis ◽  
Light Scatter ◽  
Flow Cytometric ◽  
Endogenous Fluorescence

scholarly journals The malignant cells in a Lennert's lymphoma are T lymphocytes with a mature helper surface phenotype. A multiparameter flow cytometric analysis

Blood ◽  
1986 ◽  
Vol 68 (2) ◽  
pp. 426-429 ◽  
Author(s):  
KJ Stonesifer ◽  
NA Benson ◽  
SE Ryden ◽  
DF Pawliger ◽  
RC Braylan
Keyword(s):  
Flow Cytometry ◽  
Flow Cytometric Analysis ◽  
Malignant Neoplasm ◽  
Surface Antigens ◽  
Light Scatter ◽  
Malignant Cells ◽  
T Helper ◽  
T Helper Lymphocytes ◽  
Flow Cytometric ◽  
Multiparameter Analysis

Abstract The flow cytometric analysis of DNA content in cells obtained from a case of Lennert's lymphoma demonstrated the presence of a discrete hypotetraploid cell population. Correlated multiparameter analysis of DNA, light scatter, and surface antigens by flow cytometry showed that the hypotetraploid cells were intermediate to large cells expressing T11, T3, and T4 antigens and lacking B1 and T8 antigens. These findings suggest that Lennert's lymphoma represents a malignant neoplasm of T- helper lymphocytes.

scholarly journals Flow cytometric analysis of hairy cell leukemia using right-angle light scatter

Cytometry ◽  
1986 ◽  
Vol 7 (2) ◽  
pp. 217-220 ◽  
Author(s):  
Dirk R. Van Bockstaele ◽  
Zwi N. Berneman ◽  
Marc E. Peetermans
Keyword(s):  
Hairy Cell Leukemia ◽  
Flow Cytometric Analysis ◽  
Light Scatter ◽  
Hairy Cell ◽  
Cell Leukemia ◽  
Flow Cytometric

scholarly journals The malignant cells in a Lennert's lymphoma are T lymphocytes with a mature helper surface phenotype. A multiparameter flow cytometric analysis

Blood ◽  
1986 ◽  
Vol 68 (2) ◽  
pp. 426-429 ◽  
Author(s):  
KJ Stonesifer ◽  
NA Benson ◽  
SE Ryden ◽  
DF Pawliger ◽  
RC Braylan
Keyword(s):  
Flow Cytometry ◽  
Flow Cytometric Analysis ◽  
Malignant Neoplasm ◽  
Surface Antigens ◽  
Light Scatter ◽  
Malignant Cells ◽  
T Helper ◽  
T Helper Lymphocytes ◽  
Flow Cytometric ◽  
Multiparameter Analysis

The flow cytometric analysis of DNA content in cells obtained from a case of Lennert's lymphoma demonstrated the presence of a discrete hypotetraploid cell population. Correlated multiparameter analysis of DNA, light scatter, and surface antigens by flow cytometry showed that the hypotetraploid cells were intermediate to large cells expressing T11, T3, and T4 antigens and lacking B1 and T8 antigens. These findings suggest that Lennert's lymphoma represents a malignant neoplasm of T- helper lymphocytes.

scholarly journals Actinobacillus actinomycetemcomitansLeukotoxin Induces Apoptosis in HL-60 Cells

1998 ◽  
Vol 66 (9) ◽  
pp. 4474-4483 ◽  
Author(s):  
Jonathan Korostoff ◽  
Jian Fei Wang ◽  
Irene Kieba ◽  
Mark Miller ◽  
Bruce J. Shenker ◽  
...  
Keyword(s):  
Apoptotic Cell ◽  
Critical Role ◽  
Flow Cytometric Analysis ◽  
Inflammatory Cells ◽  
Target Cells ◽  
Light Scatter ◽  
Flow Cytometric ◽  
Final Population ◽  
Cellular Debris ◽  
Phosphatidylserine Translocation

ABSTRACT Actinobacillus actinomycetemcomitans leukotoxin (Ltx) is a member of the repeats-in-toxin (RTX) family of pore-forming toxins and kills human immune cells. Currently, it remains unclear whether toxin-mediated killing of target cells involves the induction of necrosis or apoptosis. Therefore, the goal of this investigation was to determine whether Ltx is capable of causing apoptotic cell death in toxin-sensitive promyelocytic HL-60 cells. Multiparameter flow cytometric analysis of toxin-treated cells stained with Hoechst 33258 (or 33342) and 7-aminoactinomycin D allowed us to identify four populations: viable cells, early apoptotic cells, late apoptotic and/or secondarily necrotic cells, and a final population that was composed of cellular debris. Compared with control cells, HL-60 cells treated with Ltx exhibited a gradual decrease in forward light scatter with a coincident increase in side light scatter, indicative of a decrease in cell size and organelle condensation, respectively. Additional experiments demonstrated that Ltx-treated cells showed evidence of internucleosomal DNA fragmentation and phosphatidylserine translocation. The results of our studies clearly demonstrate that Ltx can kill HL-60 cells by inducing apoptosis. We hypothesize that elimination of acute inflammatory cells via this mechanism plays a critical role in the pathogenesis of diseases caused by A. actinomycetemeomitans.

Correcting Flow Cytometric Blast Counts for Blood Contamination in Bone Marrow Aspirates.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4613-4613
Author(s):  
Michael R. Loken ◽  
Sung-Chao Chu ◽  
Wayne K. Fritschle ◽  
Dian-Kun Li ◽  
Denise A. Wells
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Flow Cytometric Analysis ◽  
Myeloid Cells ◽  
Light Scatter ◽  
Blood Contamination ◽  
Simple Method ◽  
Flow Cytometric ◽  
Biopsy Specimens ◽  
Blast Count

Abstract An accurate blast count is pivotal in the diagnosis, classification and prognosis of patients with myelodysplasia. Blast counts in all previous classification schemes are based on morphologic assessment of marrow aspirates with a poor correlation to blast counts determined by flow cytometry. A significant problem in blast enumeration by flow cytometry is the variable hemodilution of the marrow during collection for flow cytometric analysis. Blast counts can vary depending on which aspirate tube is used for flow analysis, e.g., 2.4%, 1st 5ml tube; 0.62%, 2nd 5ml tube; 0.58% 3rd 5ml tube. Morphologists circumvent this problem by selecting a region for assessment close to a spicule with minimal blood dilution. Cell surface antigens can be used to distinguish mature cells found in blood as distinct from immature cells identified in marrow. CD16 intensity on neutrophils reaches a maximum at the band/segmented stage of development with a low coefficient of variation, thereby becoming a marker for mature myeloid forms. A simple method to distinguish immature from mature myeloid cells was developed to assess extent of blood contamination in marrow aspirates using a combination of CD16, CD13, and CD45. The average mature neutrophil content of a marrow was determined from phenotypically normal bone marrow biopsy specimens, assumed to have minimal blood contamination. The proportion of dimCD16 cells gated on the myeloid cells based on CD45 and right angle light scatter in 31 biopsy specimens was 82% (range 69–93, SD=6.2) (Figure 1). A value of 80% (rather than 82%) was used for the subsequent calculations to correct for the excess mature neutrophils found in an aspirate as compared to the biopsies (Corrected Blasts = [80 / % dim CD16 myeloid] x determined blast count). To test this hypothesis bone marrow aspirates were diluted with blood at different ratios to mimic blood marrow hemodilution. Blasts (defined as CD45 dim, low right angle light scatter, HLA-DR positive, CD11b negative) were determined for the various dilutions, then corrected based solely on the proportion of dim CD16 myeloid cells (Figure 2). A marrow from an MDS case was also diluted (1:5 v/v) with blood for comparison. The original marrow contained 80% dim CD16 myeloid cells with a blast count of 9.2%. After dilution, only 12% dim CD16 cells were detected with 1.1% blasts, however upon correction (6.67), the blast count was 7.3%, close to the original determination. This approach may provide for more standardization and consistency in the determination of blast counts in MDS marrow specimens using flow cytometric analysis. Figure 1, CD16 of marrow myeloid cells. Figure 1,. CD16 of marrow myeloid cells. Figure 2, Uncorrected/Corrected Blast Count Figure 2,. Uncorrected/Corrected Blast Count

scholarly journals Flow Cytometric Analysis of Microsporidia Belonging to the Genus Encephalitozoon

1999 ◽  
Vol 37 (2) ◽  
pp. 371-375 ◽  
Author(s):  
Delynn M. Moss ◽  
Gian P. Croppo ◽  
Sara Wallace ◽  
Govinda S. Visvesvara
Keyword(s):  
Monoclonal Antibody ◽  
Flow Cytometry ◽  
Polyclonal Antibodies ◽  
Flow Cytometric Analysis ◽  
Rabbit Antiserum ◽  
Fluorescein Isothiocyanate ◽  
Epidemiologic Studies ◽  
Cross Reactivity ◽  
Light Scatter ◽  
Flow Cytometric

Flow cytometry was used in the identification of human microsporidia belonging to the genus Encephalitozoon. Microsporidian spores of Encephalitozoon hellem, E. cuniculi, and E. intestinalis were propagated in axenic cultures of monkey kidney E6 cells, purified with Percoll, and exposed to homologous and heterologous rabbit antiserum and monoclonal antibody prepared against E. hellem spores. After reaction to goat anti-rabbit immunoglobulin G (IgG) or goat anti-mouse IgG conjugated to fluorescein isothiocyanate, fluorescence histograms from gated data on light-scatter profiles showed that rabbit anti-E. hellem serum was reactive to E. hellem spores but also had cross-reactivity to spores of E. cuniculi andE. intestinalis. On the other hand, fluorescence histograms showed that rabbit anti-E. cuniculi and rabbit anti-E. intestinalis sera were reactive with homologous spores only. Monoclonal antibody prepared against E. hellemreacted only with spores of E. hellem. Neither the polyclonal antibodies nor the monoclonal antibodies reacted withCryptosporidium parvum oocysts. Fluorescence histograms of spores treated with 10% formalin also showed reactivity, but the number of events in the most intense peaks of fluorescence was fewer (7 to 42%, depending on species) than the number of events in the most intense peaks of fluorescence for nontreated spores. By flow cytometry, formalin-treated and nontreated spores of Encephalitozoonwere identified to the species level by using gated data on light-scatter profiles and analyzing the fluorescence histograms from the indirect immunofluorescence of the spores. Once a procedure is established for the isolation of Encephalitozoon spores from clinical specimens, identification of spores by flow cytometry may be useful not only for diagnosis but also for epidemiologic studies.

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