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Related Concept Videos

Simple Staining Technique01:24

Simple Staining Technique

OverviewStaining techniques in microscopy enhance the visualization of microorganisms by increasing contrast and allowing the differentiation of cellular structures. Simple staining is one of the fundamental methods used to observe the basic morphological characteristics of microorganisms, including their size, shape, and arrangement. This method relies on the application of a single dye to stain the entire cell, producing a clear contrast between the cell and the background.FixationFixation is...

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A Method of Targeted Cell Isolation via Glass Surface Functionalization
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A Method of Targeted Cell Isolation via Glass Surface Functionalization

Published on: September 20, 2016

Counting unstained, confluent cells by modified bright-field microscopy.

L Louis Drey1, Michael C Graber, Jan Bieschke

  • 1Department of Biomedical Engineering, Washington University in St. Louis, St. Louis, MO, USA.

Biotechniques
|July 10, 2013
PubMed
Summary

A simple bright-field microscopy technique provides high-contrast cell images for automatic counting. This method eliminates the need for cell staining or modification, offering accurate cell counts comparable to fluorescence imaging.

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Automated Quantification and Analysis of Cell Counting Procedures Using ImageJ Plugins
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Automated Quantification and Analysis of Cell Counting Procedures Using ImageJ Plugins

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Related Experiment Videos

Last Updated: May 9, 2026

A Method of Targeted Cell Isolation via Glass Surface Functionalization
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A Method of Targeted Cell Isolation via Glass Surface Functionalization

Published on: September 20, 2016

Visualizing the Actin and Microtubule Cytoskeletons at the B-cell Immune Synapse Using Stimulated Emission Depletion (STED) Microscopy
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Visualizing the Actin and Microtubule Cytoskeletons at the B-cell Immune Synapse Using Stimulated Emission Depletion (STED) Microscopy

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Automated Quantification and Analysis of Cell Counting Procedures Using ImageJ Plugins
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Automated Quantification and Analysis of Cell Counting Procedures Using ImageJ Plugins

Published on: November 17, 2016

Area of Science:

  • Cell Biology
  • Microscopy Imaging

Background:

  • Accurate cell counting is crucial for biological research.
  • Traditional methods often require specialized equipment or cell staining.

Purpose of the Study:

  • To develop a simple, cost-effective method for high-contrast cell imaging and counting.
  • To enable routine cell quantification using standard bright-field microscopy.

Main Methods:

  • Utilizing a color filter and pinhole aperture with bright-field microscopy to generate defocused, high-contrast cell images.
  • Employing free image-analysis software for robust, automatic cell counting.
  • Benchmarking against fluorescence-based cell nucleus counting.

Main Results:

  • The technique yields high-contrast images of adherent, confluent cells (e.g., human neuroblastoma SH-EP cells).
  • Automatic cell counting from single bright-field images demonstrated high accuracy, with less than 5% error compared to fluorescence methods.
  • Cell counting accuracy was consistent across various cell densities and focal positions.

Conclusions:

  • This novel bright-field microscopy approach offers a simple, efficient, and cost-effective alternative for routine cell counting.
  • The method requires no cell-line modification or staining, making it widely accessible for biological research.