Archivos de medicina veterinaria
versión impresa ISSN 0301-732X
HARTEL, S et al. Di- and triploid erythrocyte identification by multi-parameter image analysis: A new method for the quantification of triploidization rates in rainbow trout (Oncorhynchus mykiss). Arch. med. vet. [online]. 2005, vol.37, n.2, pp. 147-154. ISSN 0301-732X. http://dx.doi.org/10.4067/S0301-732X2005000200009.
Growing international competition is forcing salmon farmers to incorporate innovative techniques into the production process. The use of triploid, all-female breeding populations offers multiple advantages over diploid populations. Currently, an exact, simple, and non- hazardous method for the quantification of diploid- and triploid salmon erythrocytes does not exist. We present a method that combines a standard microscopic bright field technique (contrast staining with GIEMSA) with multi-parameter image analysis and termed it quantitative morphologic microscopy (QMM). We used flow cytometry (FC) as the reference method to determine the DNA content of di- and triplod erythrocytes from immature rainbow trout (Oncorhynchus mykiss). Additionally, we applied quantitative fluorescence microscopy (QFM), using the DNA stains 4',6-diamidino-2-phenylindole (DAPI), propidium iodide (PI), and acridine orange (AO). Our data show that QMM possess comparable or even superior discriminating capacities than FC or QFM. The developed method opens a perspective for the classification of microscopic objects with many possible applications
Palabras llave : triploidization; cytometry; microscopy; image processing.