Reduced Fluorescence versus Forward Scatter Time-of-Flight and Increased Peak versus Integral Fluorescence Ratios Indicate Receptor Clustering in Flow Cytometry

AimsIn recent years, multiparameter flow cytometry has been increasingly recognised as an important tool in diagnosis of myelodysplastic syndrome and acute myeloid leukaemia (AML). Assessment of myeloid and monocytic ‘immunophenotypic’ dysplasia by flow cytometry in de novo AML has not been evaluated.Methods97 cases of de novo AML cases were identified and reviewed by three hematopathologists. ‘Immunophenotypic’ dysplasia was assessed on blasts, monocytes and granulocytes by mean fluorescence intensity.ResultsUsing the 2008 WHO classification criteria, there were 53 AML-not otherwise specified (NOS) (55%) and 28 AML with myelodysplasia-related changes (AML-MRC) (29%), while 16 cases were ambiguous as to AML-MRC status due to limited maturing cells for morphologic but adequate events number for immunophenotypic evaluation (AML-not evaluable, 16%). Compared with AML-NOS, granulocytic cells in AML-MRC had higher CD33 expression but lower CD45, CD11b and CD15. Monocytes in AML-MRC had lower expression of CD14, CD56 and CD45. Morphologic dysplasia was associated with significantly lower granulocytic forward scatter, side scatter and CD10 but higher CD33 expression.ConclusionsOur results suggest that the workup of AML cases should include flow cytometric assessment of granulocytes and monocytes. This analysis can aid a morphologic impression of multilineage dysplasia in distinguishing AML-MRC from AML-NOS, especially in cases with limited maturing myeloid cells.

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Assessment of deformation of human red blood cells in flow cytometry: measurement and simulation of bimodal forward scatter distributions

Biomedical Optics Express ◽

10.1364/boe.10.004531 ◽

2019 ◽

Vol 10 (9) ◽

pp. 4531

Author(s):

Jonas Gienger ◽

Hermann Gross ◽

Volker Ost ◽

Markus Bär ◽

Jörg Neukammer

Keyword(s):

Flow Cytometry ◽

Red Blood Cells ◽

Blood Cells ◽

Forward Scatter ◽

Human Red Blood Cells

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Imaging flow cytometry reveals a subset of TdT negative T‐ALL blasts with very low forward scatter on conventional flow cytometry

Cytometry Part B Clinical Cytometry ◽

10.1002/cyto.b.22035 ◽

2021 ◽

Author(s):

Carina A. Rosenberg ◽

Marie Bill ◽

Orla Maguire ◽

Marianne A. Petersen ◽

Eigil Kjeldsen ◽

...

Keyword(s):

Flow Cytometry ◽

Forward Scatter ◽

Imaging Flow Cytometry

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Comparison of electronic volume and forward scatter principles of cell selection using flow cytometry for the evaluation of acrosome and plasma membrane integrity of bull spermatozoa

Cytometry Part A ◽

10.1002/cyto.a.22474 ◽

2014 ◽

Vol 85 (8) ◽

pp. 719-728 ◽

Cited By ~ 9

Author(s):

Fride Berg Standerholen ◽

Frøydis Deinboll Myromslien ◽

Elisabeth Kommisrud ◽

Erik Ropstad ◽

Karin Elisabeth Waterhouse

Keyword(s):

Flow Cytometry ◽

Plasma Membrane ◽

Membrane Integrity ◽

Cell Selection ◽

Forward Scatter ◽

Plasma Membrane Integrity

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Forward-scatter and side-scatter dataset for epithelial cells from touch samples analyzed by flow cytometry

Data in Brief ◽

10.1016/j.dib.2015.12.027 ◽

2016 ◽

Vol 6 ◽

pp. 416-418 ◽

Cited By ~ 2

Author(s):

Cristina E. Stanciu ◽

Ye Jin Kwon ◽

Christopher J. Ehrhardt

Keyword(s):

Flow Cytometry ◽

Epithelial Cells ◽

Forward Scatter

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COVID-19 infection induces readily detectable morphological and inflammation-related phenotypic changes in peripheral blood monocytes, the severity of which correlate with patient outcome

10.1101/2020.03.24.20042655 ◽

2020 ◽

Cited By ~ 41

Author(s):

Dan Zhang ◽

Rui Guo ◽

Lei Lei ◽

Hongjuan Liu ◽

Yawen Wang ◽

...

Keyword(s):

Patient Outcome ◽

Flow Cytometry ◽

Peripheral Blood ◽

Flow Cytometric Analysis ◽

Blood Monocytes ◽

Forward Scatter ◽

Inflammatory Monocytes ◽

Flow Cytometric ◽

Functional Differences ◽

Inflammatory Phenotype

AbstractBackgroundExcessive monocyte/macrophage activation with the development of a cytokine storm and subsequent acute lung injury, leading to acute respiratory distress syndrome (ARDS) is a feared consequence of infection with COVID-19. The ability to recognize and potentially intervene early in those patients at greatest risk of developing this complication could be of great clinical utility.MethodsWe performed detailed flow cytometric analysis of peripheral blood samples from 28 COVID-19 patients treated at Xian No.8 Hospital and the First Affiliated Hospital of Xian Jiaotong University in early 2020 in an attempt to identify factors that could help predict severity of disease and patient outcome.FindingsWhile we did not detect significant differences in the number of monocytes between patients with COVID-19 and normal healthy individuals,we did identify significant morphological and functional differences, which are more pronounced in patients requiring prolonged hospitalization and ICU admission. Patients with COVID-19 have larger than normal monocytes, easily identified on forward scatter, side scatter analysis by routine flow cytometry,with the presence of a distinct population of monocytes with high forward scatter (FSC-high). On more detailed analysis, these FSC-high monocytes are CD11b+, CD14+, CD16+, CD68+, CD80+, CD163+, CD206+ and secrete IL-6, IL-10 and TNF-alpha, consistent with an inflammatory phenotype.ConclusionsThe detection and serial monitoring of this subset of inflammatory monocytes using flow cytometry could be of great help in guiding the prognostication and treatment of patients with COVID-19 and merits further evaluation.

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Flow cytometry dataset for cells collected from touched surfaces

F1000Research ◽

10.12688/f1000research.8338.2 ◽

2016 ◽

Vol 5 ◽

pp. 390

Author(s):

Ye Jin Kwon ◽

Cristina E. Stanciu ◽

M. Katherine Philpott ◽

Christopher J. Ehrhardt

Keyword(s):

Flow Cytometry ◽

Optical Properties ◽

Genetic Analysis ◽

Human Leukocyte ◽

Cell Populations ◽

Single Source ◽

Forward Scatter ◽

Physical Separation ◽

Leukocyte Antigen ◽

Genetic Profiles

‘Touch’ or trace cell mixtures submitted as evidence are a significant problem for forensic laboratories as they can render resulting genetic profiles difficult or even impossible to interpret. Optical signatures that distinguish epidermal cell populations from different contributors could facilitate the physical separation of mixture components prior to genetic analysis, and potentially the downstream production of single source profiles and/or simplified mixtures. This dataset comprises the results from antibody hybridization surveys using Human Leukocyte Antigen (HLA) and Cytokeratin (CK) probes, as well as surveys of optical properties of deposited cells, including forward scatter (FSC), side scatter (SSC), and fluorescence emissions in the Allophycocyanin (APC) channel. All analyses were performed on “touch” samples deposited by several different contributors on multiple days to assess inter- and intra-contributor variability.

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Loss of Zyx Impairs Neutrophil Production in Zebrafish Embryos

Blood ◽

10.1182/blood-2019-126644 ◽

2019 ◽

Vol 134 (Supplement_1) ◽

pp. 5001-5001

Author(s):

Katsuhiro Konno ◽

Kasem Kulkeaw ◽

Takenobu Nii ◽

Ayako Kaneyuki ◽

Daisuke Sugiyama

Keyword(s):

Flow Cytometry ◽

Functional Analysis ◽

Conflicts Of Interest ◽

Zebrafish Embryos ◽

Myeloperoxidase Activity ◽

Adult Zebrafish ◽

Forward Scatter ◽

Zebrafish Larvae ◽

Sudan Black B ◽

Adult Kidney

Zebrafish is a useful model to study vertebrate hematopoiesis, but lack of antibodies to zebrafish proteins has limited purification of hematopoietic cells. Here, we purified neutrophils from larval and adult zebrafish using the lectin Phaseolus vulgaris erythroagglutinin (PHA-E) and DRAQ5, a DNA-staining fluorescent dye. In adult kidney marrow, we purified neutrophil-like PHA-E4low DRAQ5low cells, which neutrophil-type granules. Specifically, at 96-hr post fertilization, we sorted large-sized cells from larvae using forward scatter and found that they consisted of PHA-Elow DRAQ5low populations. These cells had myeloperoxidase activity, were Sudan Black B-positive and expressed high levels of neutrophil-specific (csf3r and mpx) mRNAs, all neutrophil characteristics. Using this method, we conducted functional analysis suggesting that zyxin (zyx) plays a role in neutrophil generation in zebrafish larvae. Overall, PHA-E and DRAQ5-based flow cytometry may serve as a tool to purify zebrafish neutrophils. Disclosures No relevant conflicts of interest to declare.

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Loss of Histochemical Identity in Mast Cells Lacking Carboxypeptidase A

Molecular and Cellular Biology ◽

10.1128/mcb.25.14.6199-6210.2005 ◽

2005 ◽

Vol 25 (14) ◽

pp. 6199-6210 ◽

Cited By ~ 62

Author(s):

Thorsten B. Feyerabend ◽

Heinz Hausser ◽

Annette Tietz ◽

Carmen Blum ◽

Lars Hellman ◽

...

Keyword(s):

Flow Cytometry ◽

Mast Cell ◽

Mast Cells ◽

Alcian Blue ◽

Cell Phenotype ◽

Carboxypeptidase A ◽

Forward Scatter ◽

Peritoneal Mast Cells

ABSTRACT Mast cell carboxypeptidase A (Mc-cpa) is a highly conserved secretory granule protease. The onset of expression in mast cell progenitors and lineage specificity suggest an important role for Mc-cpa in mast cells. To address the function of Mc-cpa, we generated Mc-cpa-null mice. Mc-cpa − / − mast cells lacked carboxypeptidase activity, revealing that Mc-cpa is a nonredundant enzyme. While Mc-cpa − / − peritoneal mast cells were ultrastructurally normal and synthesized normal amounts of heparin, they displayed striking histochemical and biochemical hallmarks of immature mast cells. Wild-type peritoneal mast cells had a mature phenotype characterized by differential histochemical staining with proteoglycan-reactive dyes (cells do not stain with alcian blue but stain with safranin and with berberine) and a high side scatter to forward scatter ratio by flow cytometry and were detergent resistant. In contrast, Mc-cpa − / − peritoneal mast cells, like immature bone marrow-derived cultured mast cells, stained with alcian blue normally or weakly and either did not stain with safranin and berberine or stained weakly, had a low side scatter to forward scatter ratio, and were detergent sensitive. This phenotype was partially ameliorated with age. Thus, histochemistry and flow cytometry, commonly used to measure mast cell maturation, deviated from morphology in Mc-cpa − / − mice. The Mc-cpa − / − mast cell phenotype was not associated with defects in degranulation in vitro or passive cutaneous anaphylaxis in vivo. Collectively, Mc-cpa plays a crucial role for the generation of phenotypically mature mast cells.

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