TY - JOUR
T1 - Fluorescent Protein-Based Cellular Assays Analyzed by Laser-Scanning Microplate Cytometry in 1536-Well Plate Format
AU - Auld, Douglas S.
AU - Johnson, Ronald L.
AU - Zhang, Ya qin
AU - Veith, Henrike
AU - Jadhav, Ajit
AU - Yasgar, Adam
AU - Simeonov, Anton
AU - Zheng, Wei
AU - Martinez, Elisabeth D.
AU - Westwick, John K.
AU - Austin, Christopher P.
AU - Inglese, James
PY - 2006
Y1 - 2006
N2 - Microtiter plate readers have evolved from photomultiplier and charged-coupled device-based readers, where a population-averaged signal is detected from each well, to microscope-based imaging systems, where cellular characteristics from individual cells are measured. For these systems, speed and ease of data analysis are inversely proportional to the amount of data collected from each well. Microplate laser cytometry is a technology compatible with a 1536-well plate format and capable of population distribution analysis. Microplate cytometers such as the Acumen Explorer can monitor up to four fluorescent signals from single objects in microtiter plates with densities as high as 1536 wells. These instruments can measure changes in fluorescent protein expression, cell shape, or simple cellular redistribution events such as cytoplasmic to nuclear translocation. To develop high-throughput screening applications using laser-scanning microplate cytometry, we used green fluorescent protein- and yellow fluorescent protein-expressing cell lines designed to measure diverse biological functions such as nuclear translocation, epigenetic signaling, and G protein-coupled receptor activation. This chapter illustrates the application of microplate laser cytometry to these assays in a manner that is suitable for screening large compound collections in high throughput.
AB - Microtiter plate readers have evolved from photomultiplier and charged-coupled device-based readers, where a population-averaged signal is detected from each well, to microscope-based imaging systems, where cellular characteristics from individual cells are measured. For these systems, speed and ease of data analysis are inversely proportional to the amount of data collected from each well. Microplate laser cytometry is a technology compatible with a 1536-well plate format and capable of population distribution analysis. Microplate cytometers such as the Acumen Explorer can monitor up to four fluorescent signals from single objects in microtiter plates with densities as high as 1536 wells. These instruments can measure changes in fluorescent protein expression, cell shape, or simple cellular redistribution events such as cytoplasmic to nuclear translocation. To develop high-throughput screening applications using laser-scanning microplate cytometry, we used green fluorescent protein- and yellow fluorescent protein-expressing cell lines designed to measure diverse biological functions such as nuclear translocation, epigenetic signaling, and G protein-coupled receptor activation. This chapter illustrates the application of microplate laser cytometry to these assays in a manner that is suitable for screening large compound collections in high throughput.
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U2 - 10.1016/S0076-6879(06)14029-X
DO - 10.1016/S0076-6879(06)14029-X
M3 - Article
C2 - 17110211
AN - SCOPUS:33750843262
SN - 0076-6879
VL - 414
SP - 566
EP - 589
JO - Methods in Enzymology
JF - Methods in Enzymology
ER -