Flow cytometry in biodiversity surveys: methods, utility, and constraints

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Abstract
Flow cytometry of blood is a powerful tool for rapidly sorting individual specimens on the basis of cellular DNA content. During biodiversity surveys, the method enabled the early identification of both cryptic sympatric and allopatric species of Vietnamese ranid frogs. This method may be extremely valuable in sorting individuals from other taxa and geographic regions, especially when cellular DNA content is known to vary among closely related taxa, and in tropical situations where crypsis is a relatively common phenomenon. Protocols for preparation of freezing solution, field procedures, preparation of reference standards, and flow cytometric analysis are provided. The best method for field preservation of blood is freezing in liquid nitrogen; field fixation of blood in ethanol was less efficient and resulted in drastically increased coefficients of variation. Once samples have been transferred to freezer storage, they should not be returned to a lower storage temperature in liquid nitrogen.

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