Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Microbial Growth Measurement: Direct Methods

Microbial Growth Measurement: Direct Methods

Direct methods for measuring microbial populations in a culture are essential tools in microbiology, providing quantitative data for various applications. Among these, microscopic counts, plate counts, and serial dilution are widely used techniques, each with unique principles and applications.Microscopic CountsMicroscopic counting involves the use of a Petroff-Hausser chamber, a specialized microscope slide with a grid and defined depth. By observing a liquid culture under a microscope,...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

AI driven 3D subcellular RPE map discovers cell state transitions in establishment of apical-basal polarity.

NPJ artificial intelligence·2026

Same author

VOC injection into a house reveals large surface reservoir sizes in an indoor environment.

Proceedings of the National Academy of Sciences of the United States of America·2025

Same author

Measurement quality metrics to improve absolute microbial cell counting.

Frontiers in microbiology·2025

Same author

Nondestructive, longitudinal, 3D oxygen imaging of cells in a multi-well plate using pulse electron paramagnetic resonance imaging.

Npj imaging..·2025

Same author

Author Correction: Nondestructive, longitudinal, 3D oxygen imaging of cells in a multi-well plate using pulse electron paramagnetic resonance imaging.

Npj imaging..·2025

Same author

Analysis of the measurements used as potency tests for the 31 US FDA-approved cell therapy products.

Journal of translational medicine·2025

Same journal

Thymidylate synthase inhibitory drugs induce p53-dependent pathways differently.

PloS one·2026

Same journal

Top-down and bottom-up attention for joint pattern classification and reconstruction.

PloS one·2026

Same journal

Short- and long-term scaling behavior of blood pressure and pulse arrival time during sleep in healthy controls and patients with obstructive sleep apnea.

PloS one·2026

Same journal

Double DQN-based secrecy energy efficiency and fairness performance in IRS-assisted NOMA systems with friendly jamming.

PloS one·2026

Same journal

10 recommendations for strengthening citizen science for improved societal and ecological outcomes: A co-produced analysis of challenges and opportunities in the 21st century.

PloS one·2026

Same journal

Paying in public: Peer effects, impression management, and willingness to pay on digital payment platforms.

PloS one·2026

See all related articles

Search research articles

Related Experiment Video

Updated: Oct 6, 2025

Viability Assays for Cells in Culture

Viability Assays for Cells in Culture

Published on: January 20, 2014

Quantitative, traceable determination of cell viability using absorbance microscopy.

Greta Babakhanova¹, Stephen M Zimmerman², Laura T Pierce¹

¹Biosystems and Biomaterials Division, Material Measurement Laboratory, National Institute of Standards and Technology, Gaithersburg, MD, United States of America.

|January 19, 2022

Summary

This summary is machine-generated.

A new traceable absorbance microscopy method quantifies intracellular trypan blue uptake for accurate cell viability assessment. This method enables reliable live-dead cell classification, crucial for cell therapy and biomanufacturing quality control.

More Related Videos

Internalization and Observation of Fluorescent Biomolecules in Living Microorganisms via Electroporation

Internalization and Observation of Fluorescent Biomolecules in Living Microorganisms via Electroporation

Published on: February 8, 2015

Fluorescence Microscopy Methods for Determining the Viability of Bacteria in Association with Mammalian Cells

Fluorescence Microscopy Methods for Determining the Viability of Bacteria in Association with Mammalian Cells

Published on: September 5, 2013

Related Experiment Videos

Last Updated: Oct 6, 2025

Viability Assays for Cells in Culture

Viability Assays for Cells in Culture

Published on: January 20, 2014

Internalization and Observation of Fluorescent Biomolecules in Living Microorganisms via Electroporation

Internalization and Observation of Fluorescent Biomolecules in Living Microorganisms via Electroporation

Published on: February 8, 2015

Fluorescence Microscopy Methods for Determining the Viability of Bacteria in Association with Mammalian Cells

Fluorescence Microscopy Methods for Determining the Viability of Bacteria in Association with Mammalian Cells

Published on: September 5, 2013

Area of Science:

Biotechnology
Cell Biology
Microscopy

Background:

Cell viability assays are critical for cell therapy products but often lack traceability.
Current trypan blue dye exclusion methods rely on subjective intensity-based classifications, hindering result comparison.
Variability in pixel intensity measurements complicates accurate live-dead cell differentiation.

Purpose of the Study:

To develop a traceable absorbance microscopy method for quantifying intracellular trypan blue uptake.
To enable accurate and reproducible cell viability measurements for biomanufacturing.
To establish a standardized method for live-dead cell classification.

Main Methods:

Developed an open-source AbsorbanceQ application to convert brightfield images to quantitative absorbance images.
Quantified moles of trypan blue per cell by analyzing absorbance values.
Validated the absorbance microscopy method using neutral density filters and across multiple microscopes.

Main Results:

Absorbance microscopy demonstrated a mean absolute deviation of 3% from expected optical density values across four microscopes.
Quantified differences in trypan blue uptake between cells killed by different methods (heat shock vs. formaldehyde).
Accurately identified dead cells in a 50/50 mixture (53% dead ±6% SD) and assessed viability in stressed cell populations.

Conclusions:

Absorbance microscopy provides traceable units (moles of trypan blue) for cell viability assessment, overcoming limitations of arbitrary intensity values.
This method enhances the reliability of live-dead cell classification, essential for quality assurance in cell manufacturing.
The developed technique offers a standardized and quantitative approach for cell viability testing in biopharmaceutical applications.