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Counting total cell numbers microscopically on stained films

C Jiang1, F Wang, T T Li

  • 1Department of Biological Science and Biotechnology, Hangzhou University, People's Republic of China.

Letters in Applied Microbiology
|April 1, 1995
PubMed
Summary
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This study presents a new method for accurate cell counting by eliminating errors in spreading and distribution. The improved technique ensures reliable results for yeast cell determinations, with coefficients of variation as low as 1.1%.

Area of Science:

  • Microbiology
  • Biotechnology
  • Cell Biology

Background:

  • Conventional stained film cell counting methods suffer from errors due to imperfect sample spreading and uneven cell distribution.
  • These inaccuracies can significantly impact the reliability of cell count results, particularly in microbiological and biotechnological applications.

Purpose of the Study:

  • To develop and validate a modified cell counting method that eliminates common sources of error in stained film preparations.
  • To improve the accuracy and reproducibility of yeast cell quantification.

Main Methods:

  • A novel technique involving the addition of staining solution and sodium alginate to cell suspensions prior to slide preparation.
  • Utilizing a microsyringe to spread precise volumes (20 microliters) of the suspension in defined bands (1.7 mm x 60 mm).

Related Experiment Videos

  • Counting all cells within the field of view at 100x magnification by systematically moving the slide.

    • Main Results:

    • The modified method successfully eliminated errors associated with imperfect spreading and uneven cell distribution.
    • High accuracy and reproducibility were demonstrated, with coefficients of variation of 2.6% and 1.1% in two separate yeast cell determinations.
    • The method allows for easy total cell counting under standard microscopy.

    Conclusions:

    • The developed cell counting technique offers a significant improvement over conventional methods for accurate and reliable cell quantification.
    • The method is particularly effective for yeast cell counting, providing precise results with minimal variation.
    • Further research into automated image analysis for higher magnifications is recommended to enhance the method's applicability.