Rapid, Precise, and Accurate Counts of Symbiodinium Cells Using the Guava Flow Cytometer, and a Comparison to Other Methods

In studies of both the establishment and breakdown of cnidarian-dinoflagellate symbiosis, it is often necessary to determine the number of Symbiodinium cells relative to the quantity of host tissue. Ideally, the methods used should be rapid, precise, and accurate. In this study, we systematically ev...

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Bibliographic Details
Published in:PloS one 2015-08, Vol.10 (8), p.e0135725-e0135725
Main Authors: Krediet, Cory J, DeNofrio, Jan C, Caruso, Carlo, Burriesci, Matthew S, Cella, Kristen, Pringle, John R
Format: Article
Language:English
Subjects:
Accuracy
Algae
Animals
Automation
Cnidaria - cytology
Counting
Dinoflagellida - cytology
Electrolytes
Flow Cytometry - methods
Fluorescence
Gene expression
Guava
Image processing
Medicine
Microorganisms
Proteins
Sample preparation
Sampling methods
Symbiodinium
Symbiosis
Symbiosis - physiology
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Summary:In studies of both the establishment and breakdown of cnidarian-dinoflagellate symbiosis, it is often necessary to determine the number of Symbiodinium cells relative to the quantity of host tissue. Ideally, the methods used should be rapid, precise, and accurate. In this study, we systematically evaluated methods for sample preparation and storage and the counting of algal cells using the hemocytometer, a custom image-analysis program for automated counting of the fluorescent algal cells, the Coulter Counter, or the Millipore Guava flow-cytometer. We found that although other methods may have value in particular applications, for most purposes, the Guava flow cytometer provided by far the best combination of precision, accuracy, and efficient use of investigator time (due to the instrument's automated sample handling), while also allowing counts of algal numbers over a wide range and in small volumes of tissue homogenate. We also found that either of two assays of total homogenate protein provided a precise and seemingly accurate basis for normalization of algal counts to the total amount of holobiont tissue.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0135725