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Cell Viability Assays

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Cover of 'Cell Viability Assays'

Table of Contents

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    Book Overview
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    Chapter 1 Basic Colorimetric Proliferation Assays: MTT, WST, and Resazurin
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    Chapter 2 Assaying Cellular Viability Using the Neutral Red Uptake Assay
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    Chapter 3 Assessment of Cell Viability with Single-, Dual-, and Multi-Staining Methods Using Image Cytometry
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    Chapter 4 High-Throughput Spheroid Screens Using Volume, Resazurin Reduction, and Acid Phosphatase Activity
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    Chapter 5 A Protocol for In Vitro High-Throughput Chemical Susceptibility Screening in Differentiating NT2 Stem Cells
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    Chapter 6 Ferroptosis and Cell Death Analysis by Flow Cytometry
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    Chapter 7 Assaying Mitochondrial Respiration as an Indicator of Cellular Metabolism and Fitness
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    Chapter 8 An ATP-Based Luciferase Viability Assay for Animal African Trypanosomes Using a 96-Well Plate
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    Chapter 9 SYBR® Green I-Based Fluorescence Assay to Assess Cell Viability of Malaria Parasites for Routine Use in Compound Screening
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    Chapter 10 Screening Applications to Test Cellular Fitness in Transwell® Models After Nanoparticle Treatment
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    Chapter 11 Assays for Analyzing the Role of Transport Proteins in the Uptake and the Vectorial Transport of Substances Affecting Cell Viability
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    Chapter 12 Metabolite Profiling of Mammalian Cell Culture Processes to Evaluate Cellular Viability
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    Chapter 13 Assaying Spontaneous Network Activity and Cellular Viability Using Multi-well Microelectrode Arrays
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    Chapter 14 Quantitative Ratiometric Ca2+ Imaging to Assess Cell Viability
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    Chapter 15 Functional Viability: Measurement of Synaptic Vesicle Pool Sizes
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    Chapter 16 Phenotyping Cellular Viability by Functional Analysis of Ion Channels: GlyR-Targeted Screening in NT2-N Cells
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    Chapter 17 Systematic Cell-Based Phenotyping of Missense Alleles
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    Chapter 18 Second Harmonic Generation Microscopy of Muscle Cell Morphology and Dynamics
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    Chapter 19 Assessment of Population and ECM Production Using Multiphoton Microscopy as an Indicator of Cell Viability
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    Chapter 20 Average Rheological Quantities of Cells in Monolayers
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    Chapter 21 Measurement of Cellular Behavior by Electrochemical Impedance Sensing
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    Chapter 22 Nano-QSAR Model for Predicting Cell Viability of Human Embryonic Kidney Cells
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    Chapter 23 Erratum to: Functional Viability: Measurement of Synaptic Vesicle Pool Sizes
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    Chapter 24 Erratum to: Phenotyping Cellular Viability by Functional Analysis of Ion Channels: GlyR-Targeted Screening in NT2-N Cells
Attention for Chapter 3: Assessment of Cell Viability with Single-, Dual-, and Multi-Staining Methods Using Image Cytometry
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Chapter title
Assessment of Cell Viability with Single-, Dual-, and Multi-Staining Methods Using Image Cytometry
Chapter number 3
Book title
Cell Viability Assays
Published in
Methods in molecular biology, January 2017
DOI 10.1007/978-1-4939-6960-9_3
Pubmed ID
Book ISBNs
978-1-4939-6959-3, 978-1-4939-6960-9

Leo Li-Ying Chan, Kelsey J. McCulley, Sarah L. Kessel


Daniel F. Gilbert, Oliver Friedrich


The ability to accurately measure cell viability is important for any cell-based assay. Traditionally, viability measurements have been performed using the trypan blue exclusion method on a hemacytometer, which allows researchers to visually distinguish viable from nonviable cells. While the trypan blue method can work for cell lines or primary cells that have been rigorously purified, in more complex samples such as PBMCs, bone marrow, whole blood, or any sample with low viability, this method can lead to errors. In recent years, advances in optics and fluorescent dyes have led to the development of automated benchtop image-based cell counters for rapid cell concentration and viability measurement. In this work, we demonstrate the use of image-based cytometry for cell viability detection using single-, dual-, or multi-stain techniques. Single-staining methods using nucleic acid stains such as EB, PI, 7-AAD, DAPI, SYTOX Green, and SYTOX Red, and enzymatic stains such as CFDA and Calcein AM, were performed. Dual-staining methods using AO/PI, CFDA/PI, Calcein AM/PI, Hoechst/PI, Hoechst/DRAQ7, and DRAQ5/DAPI that enumerate viable and nonviable cells were also performed. Finally, Hoechst/Calcein AM/PI was used for a multi-staining method. Fluorescent viability staining allows exclusion of cellular debris and nonnucleated cells from analysis, which can eliminate the need to perform purification steps during sample preparation and improve efficiency. Image cytometers increase speed and throughput, capture images for visual confirmation of results, and can greatly simplify cell count and viability measurements.

Mendeley readers

The data shown below were compiled from readership statistics for 23 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 23 100%

Demographic breakdown

Readers by professional status Count As %
Researcher 5 22%
Student > Master 5 22%
Student > Bachelor 3 13%
Student > Ph. D. Student 1 4%
Student > Doctoral Student 1 4%
Other 0 0%
Unknown 8 35%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 9 39%
Immunology and Microbiology 2 9%
Agricultural and Biological Sciences 1 4%
Veterinary Science and Veterinary Medicine 1 4%
Materials Science 1 4%
Other 1 4%
Unknown 8 35%