A comparative analysis of traditional and fluorescent spermatozoa staining techniques

• Main Author: Manolakis, Alexis
• Language: en_US
• Published: Boston University 2015
• Online Access: https://hdl.handle.net/2144/12502

Thesis (M.S.)--Boston University PLEASE NOTE: Boston University Libraries did not receive an Authorization To Manage form for this thesis or dissertation. It is therefore not openly accessible, though it may be available by request. If you are the author or principal advisor of this work and would like to request open access for it, please contact us at [e-mail]. Thank you. === The purpose of this study was to compare the traditional KPIC method for staining sperm to modern fluorescent techniques employed by SPERM HY-LITERTM. Factors such as staining time, sperm recovery, and the ability to quickly and accurately count sperm were evaluated. Dilutions ranging from 1:2 to 1:10,000,000 of semen, post-coital swabs and sperm/epithelial cell suspensions were stained with SPERM HY-LITERTM, SPERM HY-LITERTM Express and KPIC. Case work samples were also analyzed with KPIC and SPERM HY-LITERTM. The sperm counts for the more concentrated dilutions of semen and sperm/e-cell samples were statistically similar in both KPIC and SPERM HY-LITERTM. However, post-coital samples stained with SPERM HY-LITERTM were statistically different than samples stained with KPIC, resulting in the observation of fewer cells. Similar results were observed with SPERM HY-LITERTM Express as well as with case work samples stained with SPERM HY-LITERTM. Although SPERM HY-LITERTM has the benefits of quicker and easier scanning due to the fluorescent filters, it is considerably more costly to employ and did not result in greater sperm recovery. Overall, KPIC provided the best results in the shortest amount of time and within a reasonable budget.