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Microalgae Cell Counting Techniques

This document describes methods for counting microalgae cells using a haemacytometer to determine the density of microalgae cultures. A culture of Chlorella sp. and Nannochloropsis sp. was provided and 10 μl of each culture was placed on a haemacytometer and the cells in four quadrants were counted under 400x magnification. The cell densities were calculated using formulas that take into account dilution factors and chamber volumes. Triplicate counts were performed and average densities of 1.03 x 106 cells/mL for Chlorella sp. and 2.67 x 106 cells/mL for Nannochloropsis sp. were obtained. Direct cell counting using a haemacytometer allows for observing cell morphology

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123 views2 pages

Microalgae Cell Counting Techniques

This document describes methods for counting microalgae cells using a haemacytometer to determine the density of microalgae cultures. A culture of Chlorella sp. and Nannochloropsis sp. was provided and 10 μl of each culture was placed on a haemacytometer and the cells in four quadrants were counted under 400x magnification. The cell densities were calculated using formulas that take into account dilution factors and chamber volumes. Triplicate counts were performed and average densities of 1.03 x 106 cells/mL for Chlorella sp. and 2.67 x 106 cells/mL for Nannochloropsis sp. were obtained. Direct cell counting using a haemacytometer allows for observing cell morphology

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Rianie
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Introduction

Microalgae cell counting is one way to estimate the density of microalgae in a volume.
Cell number of microalgae can be determined using a haemacytometer. The
haemacytometer is suitable for cells in the range of blood cells (i.e. less than about 100
µm in diameter) and for cell densities less than 105/mL. Larger cells than 100 µm are
better counted using the Sedgwick-Rafter chamber (N.R. Moheimani, et all., 2013).

Objective :
To counting the density of microalgae culture.

Methods :
A. Estimation of Microalgae Cell Densities
1. a growing culture of Chlorella sp. and Nannochloropsis sp. will be provided.
2. a glass slide is placed on the top of the haemacytometer.
3. 10 µl of the microalgae culture is taken carefully using pipette.
4. the algal cells are counted under 400x (40x ocular) magnification.
5. algal cells suspension should be diluted enough, so that it can be counted between
100-300 algal cells per quadrant (<50 cells in one box).
6. the counts in the quadrants are taken the average and the algal density in the sample is
calculated, taking into account the dilution factor
7. for cells that overlap the quadrant borders, the cells on the top and right borders are
counted, and omit the cells on the bottom and left borders of the quadrants. (each
quadrant has an area of 1 mm2, is 0,1 mm deep and has a volume of 1 x 10-4 mL).

B. Calculation
1. the cells are counted in the 4 corner quadrants of the counting chamber.
2. the cell density (cell/mL) is calculated using the formula bellow :
i) (average no of cells below and above) x 10000 x df
ii) (total cell count) x 2500 x df
Result
Times of Microalgae Density (cell/mL)
Chlorella sp. Nannochloropsis sp.
Counting
1 1,55 x106 2,20 x 106
2 0,80 x 106 3,05 x 106
3 0,75 x 106 2,75 x 106
Average 1,03 x 106 2,67 x 106

Discussion
Counting cells of microalgae in this practical were done triplet of counting to get more
accurate number of cells and reduce human error. Sample of Chlorella sp. was diluted
for two times because it has high stocking density. By having the density of microalgae
that cultured, it can be used to calculate how many sample volume of microalgae will be
transferred into a new medium or even given to the fish, and also to determine the cells
growth of microalgae.

Direct cell counting using haemacytometer has the advantage over other methods (e.g.
optical density or using a particle counter) in that one is also closely observing the cells
and any anomalies such as changes in cell morphology and/or the presence of
contaminans will be detected easily; however it is also time-consuming (N.R.
Moheimani, et all., 2013).

Conclusion
Microalgae density is one of important parameter in culturing microalgae. It can get by
counting the cells of microalgae.

References :
N.R. Moheimani, M.A. Borowitzka, A. Isdepsky, S.F. Sing, 2013, Algae for Biofuels
and Energy; Standard methods for measuring growth of algae and their composition,
Developments in Applied Phycology 5, Springer Science and Business Media
Dordrecht.

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