Kinetics of virus adsorption to single cells using fluorescent membrane probes and multiparameter flow cytometry

1982 ◽  
Vol 4 (1) ◽  
pp. 63-75 ◽  
Author(s):  
J. F. Leary ◽  
M. F. D. Notter
Keyword(s):  
Flow Cytometry ◽  
Single Cells ◽  
Multiparameter Flow Cytometry ◽  
Virus Adsorption ◽  
Kinetics Of ◽  
Membrane Probes

scholarly journals Clinical impact of panel-based error-corrected next generation sequencing versus flow cytometry to detect measurable residual disease (MRD) in acute myeloid leukemia (AML)

Leukemia ◽  
2021 ◽  
Author(s):  
Nikhil Patkar ◽  
Chinmayee Kakirde ◽  
Anam Fatima Shaikh ◽  
Rakhi Salve ◽  
Prasanna Bhanshe ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Next Generation Sequencing ◽  
Residual Disease ◽  
Multiparameter Flow Cytometry ◽  
Next Generation ◽  
Free Survival ◽  
Additional Information ◽  
Next Generation Sequencing Ngs ◽  
Kinetics Of ◽  
Generation Sequencing

AbstractWe accrued 201 patients of adult AML treated with conventional therapy, in morphological remission, and evaluated MRD using sensitive error-corrected next generation sequencing (NGS-MRD) and multiparameter flow cytometry (FCM-MRD) at the end of induction (PI) and consolidation (PC). Nearly 71% of patients were PI NGS-MRD+ and 40.9% PC NGS-MRD+ (median VAF 0.76%). NGS-MRD+ patients had a significantly higher cumulative incidence of relapse (p = 0.003), inferior overall survival (p = 0.001) and relapse free survival (p < 0.001) as compared to NGS-MRD− patients. NGS-MRD was predictive of inferior outcome in intermediate cytogenetic risk and demonstrated potential in favorable cytogenetic risk AML. PI NGS-MRD− patients had a significantly improved survival as compared to patients who became NGS-MRD− subsequently indicating that kinetics of NGS-MRD clearance was of paramount importance. NGS-MRD identified over 80% of cases identified by flow cytometry at PI time point whereas FCM identified 49.3% identified by NGS. Only a fraction of cases were NGS-MRD− but FCM-MRD+. NGS-MRD provided additional information of the risk of relapse when compared to FCM-MRD. We demonstrate a widely applicable, scalable NGS-MRD approach that is clinically informative and synergistic to FCM-MRD in AML treated with conventional therapies. Maximum clinical utility may be leveraged by combining FCM and NGS-MRD modalities.

scholarly journals Endosome pH measured in single cells by dual fluorescence flow cytometry: rapid acidification of insulin to pH 6.

1984 ◽  
Vol 98 (5) ◽  
pp. 1757-1762 ◽  
Author(s):  
R F Murphy ◽  
S Powers ◽  
C R Cantor
Keyword(s):  
Flow Cytometry ◽  
Single Cells ◽  
Cell Suspensions ◽  
Dual Fluorescence ◽  
3T3 Cells ◽  
Specific Receptor ◽  
Cell Basis ◽  
A Cell ◽  
Kinetics Of ◽  
Cell Versus

The acidification of various ligands was measured on a cell by cell basis for cell suspensions by correlated dual fluorescence flow cytometry. Mouse 3T3 cells were incubated with a mixture of fluorescein- and rhodamine-conjugated ligands, and the ratio of fluorescein and rhodamine fluorescence was used as a measure of endosome pH. The calibration of this ratio by both fluorometry and flow cytometry is described. Dual parameter histograms of average endosome pH per cell versus amount of internalization were calculated from this data, for samples in the absence and presence of chloroquine added to neutralize acidic cellular vesicles. The kinetics of acidification of insulin were measured and compared with previous results obtained with the chloroquine ratio technique. Rapid acidification of internalized ligand was observed both for insulin, which was mostly internalized via nonspecific pathways, and for alpha 2-macroglobulin, which was mainly internalized by specific receptor-mediated endocytosis. The average pH observed for internalized insulin was less than pH 6 within 10 min after addition of insulin. At 30 min, the average pH began to decrease to approximately pH 5, presumably because of fusion of endosomes with lysosomes.

scholarly journals Clinical Impact of Panel Based Error Corrected Next Generation Sequencing versus Flow Cytometry to Detect Measurable Residual Disease (MRD) in Acute Myeloid Leukemia (AML)

2020 ◽  
Author(s):  
Nikhil Patkar ◽  
Chinmayee Kakirde ◽  
Anam Fatima Shaikh ◽  
Rakhi Salve ◽  
Prasanna Bhanshe ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Next Generation Sequencing ◽  
Myeloid Leukemia ◽  
Residual Disease ◽  
Multiparameter Flow Cytometry ◽  
Next Generation ◽  
Free Survival ◽  
Next Generation Sequencing Ngs ◽  
Kinetics Of ◽  
Generation Sequencing

We accrued 201 patients of adult AML treated with conventional therapy, in morphological remission and evaluated MRD using sensitive error corrected next generation sequencing (NGS-MRD) and multiparameter flow cytometry (FCM-MRD) at the end of induction (PI) and consolidation (PC). Nearly 71% of patients harbored PI NGS-MRD and 40.9% harbored PC NGS-MRD (median VAF 0.76%). Patients harboring NGS-MRD had a significantly higher cumulative incidence of relapse (p=0.003), inferior overall survival (p=0.001) and relapse free survival (p<0.001) as compared to NGS-MRD negative patients. NGS-MRD was predictive of inferior outcome in intermediate cytogenetic risk and demonstrated potential in favorable cytogenetic risk AML. Patients who cleared PI NGS-MRD had a significantly improved survival as compared to patients who became negative subsequently indicating that kinetics of NGS-MRD clearance was of paramount importance. NGS-MRD identified over 80% of cases identified by flow cytometry at PI time point whereas FCM identified 49.3% identified by NGS. Only a fraction of cases were truly missed by NGS as compared to FCM-MRD. NGS-MRD emerged as the most important independent prognostic factor predictive of inferior outcome (p<0.001). We demonstrate a widely applicable, scalable NGS-MRD approach that is clinically informative and advantageous when compared to FCM-MRD in AML treated with conventional therapies.

scholarly journals KD determination from time-resolved experiments on live cells with LigandTracer and reconciliation with end-point flow cytometry measurements

2021 ◽  
Author(s):  
Diana Spiegelberg ◽  
Jonas Stenberg ◽  
Pascale Richalet ◽  
Marc Vanhove
Keyword(s):  
Flow Cytometry ◽  
Live Cells ◽  
Time Resolved ◽  
Surface Receptors ◽  
Full Characterization ◽  
End Point ◽  
Titration Curves ◽  
Kinetics Of

AbstractDesign of next-generation therapeutics comes with new challenges and emulates technology and methods to meet them. Characterizing the binding of either natural ligands or therapeutic proteins to cell-surface receptors, for which relevant recombinant versions may not exist, represents one of these challenges. Here we report the characterization of the interaction of five different antibody therapeutics (Trastuzumab, Rituximab, Panitumumab, Pertuzumab, and Cetuximab) with their cognate target receptors using LigandTracer. The method offers the advantage of being performed on live cells, alleviating the need for a recombinant source of the receptor. Furthermore, time-resolved measurements, in addition to allowing the determination of the affinity of the studied drug to its target, give access to the binding kinetics thereby providing a full characterization of the system. In this study, we also compared time-resolved LigandTracer data with end-point KD determination from flow cytometry experiments and hypothesize that discrepancies between these two approaches, when they exist, generally come from flow cytometry titration curves being acquired prior to full equilibration of the system. Our data, however, show that knowledge of the kinetics of the interaction allows to reconcile the data obtained by flow cytometry and LigandTracer and demonstrate the complementarity of these two methods.

scholarly journals Kinetics of in vitro natural killer activity against K562 cells as detected by flow cytometry

Cytometry ◽  
1998 ◽  
Vol 32 (4) ◽  
pp. 280-285 ◽  
Author(s):  
Loris Zamai ◽  
Adriana R. Mariani ◽  
Giorgio Zauli ◽  
Luigi Rodella ◽  
Rita Rezzani ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Natural Killer ◽  
Natural Killer Activity ◽  
K562 Cells ◽  
Killer Activity ◽  
Kinetics Of

scholarly journals Assessment of Immunotoxicity by Multiparameter Flow Cytometry

1997 ◽  
Vol 38 (1) ◽  
pp. 38-54
Author(s):  
Scott W. Burchiel ◽  
Nancy L. Kerkvliet ◽  
G. Frank Gerberick ◽  
David A. Lawrence ◽  
Gregory S. Ladics
Keyword(s):  
Flow Cytometry ◽  
Multiparameter Flow Cytometry

T2050 Immunophenotypic Characterization of the Cellular Infiltrate During Murine Experimental Colitis Using Multiparameter Flow Cytometry

2010 ◽  
Vol 138 (5) ◽  
pp. S-621
Author(s):  
Kimberly A. Zins ◽  
Tamas Ordog ◽  
Michael R. Bardsley ◽  
Gianrico Farrugia ◽  
Joseph H. Szurszewski ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Experimental Colitis ◽  
Multiparameter Flow Cytometry ◽  
Cellular Infiltrate

Immunophenotypic Study of Basophils by Multiparameter Flow Cytometry

2008 ◽  
Vol 132 (5) ◽  
pp. 813-819
Author(s):  
Xiaohong Han ◽  
Jeffrey L. Jorgensen ◽  
Archana Brahmandam ◽  
Ellen Schlette ◽  
Yang O. Huh ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Chronic Myelogenous Leukemia ◽  
Peripheral Blood ◽  
Myelogenous Leukemia ◽  
Multiparameter Flow Cytometry ◽  
Color Flow ◽  
Aberrant Expression ◽  
Flow Cytometric ◽  
Hla Dr

Abstract Context.—The immunophenotypic profile of basophils is not yet fully established, and the immunophenotypic changes in chronic myelogenous leukemia are not fully characterized. Objective.—To establish a comprehensive immunophenotypic spectrum of normal basophils and to assess the range of immunophenotypic aberrations of basophils in chronic myelogenous leukemia. Design.—Using 4-color flow cytometry, we compared the immunophenotypic profile of basophils in peripheral blood or bone marrow samples from 20 patients with no evidence of neoplasia to basophils from 15 patients with chronic myelogenous leukemia. Results.—Basophils in control cases were all positive for CD9, CD13, CD22, CD25 (dim), CD33, CD36, CD38 (bright), CD45 (dimmer than lymphocytes and brighter than myeloblasts), and CD123 (bright), and were negative for CD19, CD34, CD64, CD117, and HLA-DR. Basophils in all chronic myelogenous leukemia patients possessed 1 to 5 immunophenotypic aberrancies. The most common aberrancies were underexpression of CD38, followed by aberrant expression of CD64 and underexpression of CD123. CD34 and CD117 were present in cases with basophilic precursors. Myeloblasts showed a distinct immunophenotypic profile, as they typically expressed CD34 and CD117, showed dimmer expression (compared with basophils) of CD38, CD45, and CD123, and lacked expression of CD22. Conclusions.—Flow cytometric immunophenotyping can identify immunophenotypic aberrations of basophils in chronic myelogenous leukemia, and discriminate basophils from myeloblasts.

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