scholarly journals Broad Applications of Multi-Colour Time-Resolved Flow Cytometry

10.5772/37241 ◽  
2012 ◽  
Author(s):  
Ben J. ◽  
James S.
Keyword(s):  
Flow Cytometry ◽  
Time Resolved

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 Subcellular localization-dependent changes in EGFP fluorescence lifetime measured by time-resolved flow cytometry

2013 ◽  
Vol 4 (8) ◽  
pp. 1390 ◽  
Author(s):  
Ali Vaziri Gohar ◽  
Ruofan Cao ◽  
Patrick Jenkins ◽  
Wenyan Li ◽  
Jessica P. Houston ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Subcellular Localization ◽  
Fluorescence Lifetime ◽  
Time Resolved ◽  
Egfp Fluorescence

scholarly journals Quantifying cell densities and biovolumes of phytoplankton communities and functional groups using scanning flow cytometry, machine learning and unsupervised clustering

2018 ◽  
Author(s):  
Mridul K. Thomas ◽  
Simone Fontana ◽  
Marta Reyes ◽  
Francesco Pomati
Keyword(s):  
Machine Learning ◽  
Flow Cytometry ◽  
Functional Groups ◽  
Clustering Algorithm ◽  
Unsupervised Clustering ◽  
Learning Tools ◽  
Time Resolved ◽  
Automated Method ◽  
Phytoplankton Communities ◽  
Cell Densities

AbstractScanning flow cytometry (SFCM) is characterized by the measurement of time-resolved pulses of fluorescence and scattering, enabling the high-throughput quantification of phytoplankton morphology and pigmentation. Quantifying variation at the single cell and colony level improves our ability to understand dynamics in natural communities. Automated high-frequency monitoring of these communities is presently limited by the absence of repeatable, rapid protocols to analyse SFCM datasets, where images of individual particles are not available. Here we demonstrate a repeatable, semi-automated method to (1) rapidly clean SFCM data from a phytoplankton community by removing signals that do not belong to live phytoplankton cells, (2) classify individual cells into trait clusters that correspond to functional groups, and (3) quantify the biovolumes of individual cells, the total biovolume of the whole community and the total biovolumes of the major functional groups. Our method involves the development of training datasets using lab cultures, the use of an unsupervised clustering algorithm to identify trait clusters, and machine learning tools (random forests) to (1) evaluate variable importance, (2) classify data points, and (3) estimate biovolumes of individual cells. We provide example datasets and R code for our analytical approach that can be adapted for analysis of datasets from other flow cytometers or scanning flow cytometers.

In vivo flow cytometry and time-resolved near-IR angiography and lymphography

2007 ◽  
Author(s):  
Ekaterina I. Galanzha ◽  
Valery V. Tuchin ◽  
Robert W. Brock ◽  
Vladimir P. Zharov
Keyword(s):  
Flow Cytometry ◽  
Time Resolved ◽  
Near Ir

High-speed flow cytometry using nonlinear Brillouin imaging/sensing via time-resolved optical (BISTRO) measurements

2016 ◽  
Author(s):  
Zhaokai Meng ◽  
Charles W. Ballman ◽  
Georgi I. Petrov ◽  
Vladislav V. Yakovlev
Keyword(s):  
Flow Cytometry ◽  
High Speed ◽  
High Speed Flow ◽  
Time Resolved ◽  
Speed Flow

scholarly journals Time-resolved flow cytometry for the measurement of lanthanide chelate fluorescence: I. Concept and theoretical evaluation

Cytometry ◽  
1994 ◽  
Vol 16 (3) ◽  
pp. 187-194 ◽  
Author(s):  
Marc A. Condrau ◽  
Reto A. Schwendener ◽  
Peter Niederer ◽  
Max Anliker
Keyword(s):  
Flow Cytometry ◽  
Theoretical Evaluation ◽  
Time Resolved ◽  
Lanthanide Chelate

scholarly journals Time-resolved flow cytometry for the measurement of lanthanide chelate fluorescence: II. Instrument design and experimental results

Cytometry ◽  
1994 ◽  
Vol 16 (3) ◽  
pp. 195-205 ◽  
Author(s):  
Marc A. Condrau ◽  
Reto A. Schwendener ◽  
Marcel Zimmermann ◽  
Markus H. Muser ◽  
Urs Graf ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Instrument Design ◽  
Experimental Results ◽  
Time Resolved ◽  
Lanthanide Chelate

scholarly journals Lifetime encoding in flow cytometry for bead-based sensing of biomolecular interaction

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Daniel Kage ◽  
Katrin Hoffmann ◽  
Heike Borcherding ◽  
Uwe Schedler ◽  
Ute Resch-Genger
Keyword(s):  
Flow Cytometry ◽  
Time Domain ◽  
High Throughput Screening ◽  
Photon Counting ◽  
Clinical Diagnostics ◽  
Biomolecular Interactions ◽  
Time Resolved ◽  
Excitation Light ◽  
Intensity Measurements ◽  
The Time Domain

Abstract To demonstrate the potential of time-resolved flow cytometry (FCM) for bioanalysis, clinical diagnostics, and optically encoded bead-based assays, we performed a proof-of-principle study to detect biomolecular interactions utilizing fluorescence lifetime (LT)-encoded micron-sized polymer beads bearing target-specific bioligands and a recently developed prototype lifetime flow cytometer (LT-FCM setup). This instrument is equipped with a single excitation light source and different fluorescence detectors, one operated in the photon-counting mode for time-resolved measurements of fluorescence decays and three detectors for conventional intensity measurements in different spectral windows. First, discrimination of bead-bound biomolecules was demonstrated in the time domain exemplarily for two targets, Streptavidin (SAv) and the tumor marker human chorionic gonadotropin (HCG). In a second step, the determination of biomolecule concentration levels was addressed representatively for the inflammation-related biomarker tumor necrosis factor (TNF-α) utilizing fluorescence intensity measurements in a second channel of the LT-FCM instrument. Our results underline the applicability of LT-FCM in the time domain for measurements of biomolecular interactions in suspension assays. In the future, the combination of spectral and LT encoding and multiplexing and the expansion of the time scale from the lower nanosecond range to the longer nanosecond and the microsecond region is expected to provide many distinguishable codes. This enables an increasing degree of multiplexing which could be attractive for high throughput screening applications.

A Ruthenium Probe for Cell Viability Measurement Using Flow Cytometry, Confocal Microscopy and Time-resolved Luminescence ¶

2007 ◽  
Vol 72 (1) ◽  
pp. 28-34 ◽  
Author(s):  
M. Emilia Jiménez-Hernández ◽  
Guillermo Orellana ◽  
Francisco Montero ◽  
M. Teresa Portolés
Keyword(s):  
Flow Cytometry ◽  
Confocal Microscopy ◽  
Cell Viability ◽  
Time Resolved
Sign in / Sign up

Export Citation Format

Share Document