Establishment of an optimal hypo-osmotic swelling test by examining single spermatozoa in four different hypo-osmotic solutions

Yieh Loong Tsai, Jiaen Liu, Jairo E. Garcia, Eugene Katz, Gail Compton, Theodore A. Baramki

Research output: Contribution to journalArticlepeer-review

28 Scopus citations


In order to find an optimal hypo-osmotic swelling test (HOST) and to identify viable sperm cells from patients with asthenozoospermia for intracytoplasmic sperm injection (ICSI), we tested single motile and non-motile spermatozoa in four hypo-osmotic solutions by micromanipulation. The four solutions were: A, H2O; B, 50 mOsm NaCl; C, 150 mOsm NaCl and D, 150 mOsm sodium citrate and fructose. Eosin Y staining was then carried out for evaluation of viability of the spermatozoa after HOST. Using motile spermatozoa, no statistical difference was found in HOST-positive spermatozoa between these four solutions. There were more viable sperm cells after HOST in solutions C and D, as noted by Eosin Y staining. After non-motile spermatozoa were incubated for 1 min in the four solutions, HOST with solution C gave the best results for identification of viable sperm cells compared to the other three solutions. When motile spermatozoa were incubated in solution C or solution D for 30 min, the result of HOST with solution C (10.8% dead spermatozoa) was superior to that of solution D (49.1% dead spermatozoa). In conclusion, the HOST protocol using 150 mOsm NaCl (solution C) for 1 min yielded the best results for selection of viable spermatozoa. This procedure should be used for selection of viable spermatozoa for ICSI in patients with 100% non-motile spermatozoa.

Original languageEnglish (US)
Pages (from-to)1111-1113
Number of pages3
JournalHuman Reproduction
Issue number5
StatePublished - 1997
Externally publishedYes


  • Asthenozoospermia
  • Eosin Y staining
  • Hypo-osmotic swelling test
  • Intracytoplasmic sperm injection

ASJC Scopus subject areas

  • Reproductive Medicine
  • Obstetrics and Gynecology


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