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Comparison of six different methods to calculate cell densities
Carolina Camacho-Fernández1, David Hervás2, Alba Rivas-Sendra1,3
11COMAV - Universitat Politècnica de València, CPI, Edificio 8E - Escalera I, Camino de Vera, s/n, 46022 Valencia, Spain.
Plant Methods
|April 25, 2018
Summary
Determining optimal cell density is crucial for in vitro cultures. This study compares six cell counting methods, finding hemacytometers most practical, automated counters a balance, and flow cytometry best for reproducibility but lacking accuracy.
Area of Science:
- Cell Biology
- Plant Biotechnology
- Biotechnology
Background:
- Accurate cell density determination is critical for successful in vitro plant and animal cell cultures.
- Traditional methods like hemacytometers have limitations, with less known about alternative techniques.
- Isolated microspore culture exemplifies the need for precise cell seeding densities.
Purpose of the Study:
- To comprehensively compare six distinct methods for calculating cell densities in vitro.
- To evaluate the technical advantages and disadvantages of each method for cell counting.
- To provide guidance for selecting the most appropriate cell counting technique for specific laboratory needs.
Main Methods:
- Comparison of six cell density calculation methods: Neubauer improved hemacytometer, automated cell counter, manual counting, and three flow cytometry techniques (autofluorescence, propidium iodide staining, side scattered light).
- Experiments utilized isolated eggplant microspore cultures and fluorescent beads (fluorospheres).
Main Results:
- Hemacytometers were found to be the most practical and widely applicable method for cell counting.
- Automated cell counters offered a balance of precision and cost but had limited accuracy.
- Flow cytometry methods demonstrated superior reproducibility and inter-method agreement but lacked accuracy and precision.
Conclusions:
- The study provides a thorough technical evaluation of various cell counting methods for in vitro cultures.
- Results offer clear criteria for laboratories to choose the most suitable counting technique.
- Findings are applicable to microspore culture, other plant cell cultures, and particle counting procedures.
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