BIOMAT Database Logo BIOMAT Database Logo

High-speed multiparameter photophysical analyses of fluorophore libraries. 2015

Kevin M Dean, and Lloyd M Davis, and Jennifer L Lubbeck, and Premashis Manna, and Pia Friis, and Amy E Palmer, and Ralph Jimenez
†BioFrontiers Institute, University of Colorado, Boulder, Colorado 80309, United States.

There is a critical need for high-speed multiparameter photophysical measurements of large libraries of fluorescent probe variants for imaging and biosensor development. We present a microfluidic flow cytometer that rapidly assays 10(4)-10(5) member cell-based fluorophore libraries, simultaneously measuring fluorescence lifetime and photobleaching. Together, these photophysical characteristics determine imaging performance. We demonstrate the ability to resolve the diverse photophysical characteristics of different library types and the ability to identify rare populations.

UI MeSH Term Description Entries
D005434 Flow Cytometry Technique using an instrument system for making, processing, and displaying one or more measurements on individual cells obtained from a cell suspension. Cells are usually stained with one or more fluorescent dyes specific to cell components of interest, e.g., DNA, and fluorescence of each cell is measured as it rapidly transverses the excitation beam (laser or mercury arc lamp). Fluorescence provides a quantitative measure of various biochemical and biophysical properties of the cell, as well as a basis for cell sorting. Other measurable optical parameters include light absorption and light scattering, the latter being applicable to the measurement of cell size, shape, density, granularity, and stain uptake. Cytofluorometry, Flow,Cytometry, Flow,Flow Microfluorimetry,Fluorescence-Activated Cell Sorting,Microfluorometry, Flow,Cell Sorting, Fluorescence-Activated,Cell Sortings, Fluorescence-Activated,Cytofluorometries, Flow,Cytometries, Flow,Flow Cytofluorometries,Flow Cytofluorometry,Flow Cytometries,Flow Microfluorometries,Flow Microfluorometry,Fluorescence Activated Cell Sorting,Fluorescence-Activated Cell Sortings,Microfluorimetry, Flow,Microfluorometries, Flow,Sorting, Fluorescence-Activated Cell,Sortings, Fluorescence-Activated Cell
D005456 Fluorescent Dyes Chemicals that emit light after excitation by light. The wave length of the emitted light is usually longer than that of the incident light. Fluorochromes are substances that cause fluorescence in other substances, i.e., dyes used to mark or label other compounds with fluorescent tags. Flourescent Agent,Fluorescent Dye,Fluorescent Probe,Fluorescent Probes,Fluorochrome,Fluorochromes,Fluorogenic Substrates,Fluorescence Agents,Fluorescent Agents,Fluorogenic Substrate,Agents, Fluorescence,Agents, Fluorescent,Dyes, Fluorescent,Probes, Fluorescent,Substrates, Fluorogenic
D006367 HeLa Cells The first continuously cultured human malignant CELL LINE, derived from the cervical carcinoma of Henrietta Lacks. These cells are used for, among other things, VIRUS CULTIVATION and PRECLINICAL DRUG EVALUATION assays. Cell, HeLa,Cells, HeLa,HeLa Cell
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D013050 Spectrometry, Fluorescence Measurement of the intensity and quality of fluorescence. Fluorescence Spectrophotometry,Fluorescence Spectroscopy,Spectrofluorometry,Fluorescence Spectrometry,Spectrophotometry, Fluorescence,Spectroscopy, Fluorescence
D056656 Lab-On-A-Chip Devices Microdevices that combine microfluidics technology with electrical and/or mechanical functions for analyzing very small fluid volumes. They consist of microchannels etched into substrates made of silicon, glass, or polymer using processes similar to photolithography. The test fluids in the channels can then interact with different elements such as electrodes, photodetectors, chemical sensors, pumps, and valves. Microchip Analytical Devices,Microfluidic Devices,Microfluidic Lab-On-A-Chip,Microfluidic Microchips,Nanochip Analytical Devices,Analytical Device, Microchip,Analytical Device, Nanochip,Analytical Devices, Microchip,Analytical Devices, Nanochip,Device, Lab-On-A-Chip,Device, Microchip Analytical,Device, Microfluidic,Device, Nanochip Analytical,Devices, Lab-On-A-Chip,Devices, Microchip Analytical,Devices, Microfluidic,Devices, Nanochip Analytical,Lab On A Chip Devices,Lab-On-A-Chip Device,Lab-On-A-Chip, Microfluidic,Lab-On-A-Chips, Microfluidic,Microchip Analytical Device,Microchip, Microfluidic,Microchips, Microfluidic,Microfluidic Device,Microfluidic Lab On A Chip,Microfluidic Lab-On-A-Chips,Microfluidic Microchip,Nanochip Analytical Device
D038761 Photobleaching Light-induced change in a chromophore, resulting in the loss of its absorption of light of a particular wave length. The photon energy causes a conformational change in the photoreceptor proteins affecting PHOTOTRANSDUCTION. This occurs naturally in the retina (ADAPTATION, OCULAR) on long exposure to bright light. Photobleaching presents problems when occurring in PHOTODYNAMIC THERAPY, and in FLUORESCENCE MICROSCOPY. On the other hand, this phenomenon is exploited in the technique, FLUORESCENCE RECOVERY AFTER PHOTOBLEACHING, allowing measurement of the movements of proteins and LIPIDS in the CELL MEMBRANE.

Related Publications

Kevin M Dean, and Lloyd M Davis, and Jennifer L Lubbeck, and Premashis Manna, and Pia Friis, and Amy E Palmer, and Ralph Jimenez
High-speed telefacsimile in libraries.
January 1983, Library technology reports,
Kevin M Dean, and Lloyd M Davis, and Jennifer L Lubbeck, and Premashis Manna, and Pia Friis, and Amy E Palmer, and Ralph Jimenez
Linear absorption tomography with velocimetry (LATV) for multiparameter measurements in high-speed flows.
October 2020, Optics express,
Kevin M Dean, and Lloyd M Davis, and Jennifer L Lubbeck, and Premashis Manna, and Pia Friis, and Amy E Palmer, and Ralph Jimenez
Optimizing fluorophore density for single virus counting: a photophysical approach.
January 2021, Methods and applications in fluorescence,
Kevin M Dean, and Lloyd M Davis, and Jennifer L Lubbeck, and Premashis Manna, and Pia Friis, and Amy E Palmer, and Ralph Jimenez
Synthesis and Photophysical Properties of λ5-Phosphinines as a Tunable Fluorophore.
February 2018, Journal of the American Chemical Society,
Kevin M Dean, and Lloyd M Davis, and Jennifer L Lubbeck, and Premashis Manna, and Pia Friis, and Amy E Palmer, and Ralph Jimenez
Capsule chemistry technology for high-speed clinical chemistry analyses.
September 1985, Clinical chemistry,
Kevin M Dean, and Lloyd M Davis, and Jennifer L Lubbeck, and Premashis Manna, and Pia Friis, and Amy E Palmer, and Ralph Jimenez
High-speed screening of combinatorial libraries by gradient packed-column supercritical fluid chromatography.
July 2000, Journal of biochemical and biophysical methods,
Kevin M Dean, and Lloyd M Davis, and Jennifer L Lubbeck, and Premashis Manna, and Pia Friis, and Amy E Palmer, and Ralph Jimenez
Hybridization analyses of arrayed cDNA libraries.
October 1991, Trends in genetics : TIG,
Kevin M Dean, and Lloyd M Davis, and Jennifer L Lubbeck, and Premashis Manna, and Pia Friis, and Amy E Palmer, and Ralph Jimenez
The Effect of Fluorophore Conjugation on Antibody Affinity and the Photophysical Properties of Dyes.
February 2018, Biophysical journal,
Kevin M Dean, and Lloyd M Davis, and Jennifer L Lubbeck, and Premashis Manna, and Pia Friis, and Amy E Palmer, and Ralph Jimenez
Speed analyses of stimulus equivalence.
May 1996, Journal of the experimental analysis of behavior,
Kevin M Dean, and Lloyd M Davis, and Jennifer L Lubbeck, and Premashis Manna, and Pia Friis, and Amy E Palmer, and Ralph Jimenez
Selection of Antibodies with Tailored Properties by Application of High-Throughput Multiparameter Fluorescence-Activated Cell Sorting of Yeast-Displayed Immune Libraries.
October 2018, Molecular biotechnology,
Copied contents to your clipboard!