Comparison of three staining techniques for the morphometric study of rainbow trout ( Oncorhynchus mykiss) spermatozoa

• Published in: Theriogenology Vol. 69; no. 8; pp. 1033 - 1038
• Main Authors: Tuset, V.M., Dietrich, G.J., Wojtczak, M., Słowińska, M., de Monserrat, J., Ciereszko, A.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.05.2008; [Oxford]: Butterworth-Heinemann; [New York]: Elsevier Science
• Subjects: analytical kits; Animals; automation; Coloring Agents; differential staining; Histocytochemistry - methods; Histocytochemistry - veterinary; image analysis; Image Processing, Computer-Assisted; integrated semen analysis system; ISAS; Male; methodology; milt; Morphometry; Oncorhynchus mykiss; Rainbow trout; sperm head; Sperm Head - chemistry; sperm measurements; sperm morphology; sperm tail; Sperm Tail - chemistry; Spermatozoa; Spermatozoa - chemistry; Spermatozoa - cytology; Spermatozoa - ultrastructure; Staining and Labeling - methods; Staining and Labeling - veterinary; Staining Techniques; Trout; Spermatozoa; Staining Techniques; Morphometry; ISAS; Rainbow trout
• ISSN: 0093-691X; 1879-3231
• DOI: 10.1016/j.theriogenology.2008.01.012

This study was designed to compare the performance of the kits Diff-Quick, Hemacolor and Spermac for staining the spermatozoa of rainbow trout. Automated sperm morphology analysis (ASMA) was performed using two image analysis programs to determine the sperm measurements: head size (length, width, area and perimeter), shape (ellipticity, rugosity, elongation and regularity) and tail length. Diff-Quick was found to be the best procedure for staining the trout spermatozoa. The use of this method rendered the highest number of cells correctly analyzed, and provided good colour intensity and contrast of the sperm head. No differences among the methods were detected in terms of tail length measurements. Mean values established using Diff-Quick for the main morphometric variables were: head length 2.93 ± 0.13 μm; head width 2.33 ± 0.15 μm and tail length 34.16 ± 1.66 μm. Based on these findings, we recommend the Diff-Quick staining kit for its accurate and reproducible morphometric results. Notwithstanding, when analyzing the sperm tail of the rainbow trout, the Spermac method offers improved contrast.