Bibliographic Citation
| Document |  550 K |
|---|---|
| Title | Critical tonicity determination of sperm using fluorescent staining and flow cytometry |
| Creator/Author | Noiles, E.E. ; Ruffing, N.A. ; Kleinhans, F.W. ; Mark, L.A. ; Watson, P.F. ; Critser, J.K. (Methodist Hospital, Indianapolis, IN (USA)) ; Horstman, L. (Purdue Univ., Lafayette, IN (USA). School of Veterinary Medicine) ; Mazur, P. (Oak Ridge National Lab., TN (USA)) |
| Publication Date | 1990 Jan 01 |
| OSTI Identifier | OSTI ID: 6759774; Legacy ID: DE91000617 |
| Report Number(s) | CONF-9008158-1 |
| DOE Contract Number | AC05-84OR21400 |
| Other Number(s) | Other: ON: DE91000617 |
| Resource Type | Conference |
| Specific Type | Technical Report |
| Resource Relation | Conference: 2. international conference on the preservation of boar semen, Beltsville, MD (USA), 19-22 Aug 1990 |
| Research Org | Oak Ridge National Lab., TN (USA) |
| Sponsoring Org | DOE/ER |
| Subject | 59 BASIC BIOLOGICAL SCIENCES; CRYOGENICS; RESEARCH PROGRAMS; SPERMATOZOA; LYSIS; ACTIVATION ENERGY; CATTLE; CELL FLOW SYSTEMS; CELL KILLING; CELL MEMBRANES; CRYOBIOLOGY; EFFICIENCY; FLUORESCENCE; MAMMALS; MAN; PRODUCTIVITY; STAINS; SWELLING; TEMPERATURE EFFECTS; ANIMALS; BIOLOGY; CELL CONSTITUENTS; DOMESTIC ANIMALS; ENERGY; GAMETES; GERM CELLS; LUMINESCENCE; MEMBRANES; PRIMATES; RUMINANTS; VERTEBRATES |
| Description/Abstract | The use of cryopreserved, rather than fresh, mammalian semen for artificial insemination confers several important medical and/or economic advantages. However, current methods for cryopreservation of both human and bovine spermatozoa result in approximately only a 50% survival rate with thawing, obviously reducing the fertilizing capacity of the semen. A primary consideration during the cooling process is to avoid intracellular ice crystal formation with its lethal consequences to the cell. Current techniques achieve this by controlling the cooling rate. Computation of the time necessary for this dehydration, and hence, the cooling rate, is dependent upon knowledge of the water permeability coefficient (L{sub {rho}}) and its activation energy. The fluorophore, 6-carboxyfluoroscein diacetate (CFDA), which is nonfluorescent, readily crosses the intact plasma membrane. Intracellular esterases hydrolyze CFDA to 6-carboxyfluoroscein, a fluorescent, membrane-impermeable fluorophore. Consequently, spermatozoa with intact plasma membranes fluoresce bright green (Garner et. al., 1986), but those with disrupted membranes do not. Therefore, the purpose of this study was to use loss of CFDA fluorescence to determine the osmolality at which 50% of the spermatozoa will swell and lyse (critical tonicity, CT). These data will then be used to determine the L{sub {rho}} and its activation energy for sperm, thus increasing the knowledge available in cellular cryopreservation. 15 refs., 3 figs. |
| Country of Publication | United States |
| Language | English |
| Format | Medium: ED; Size: Pages: (6 p) |
| Availability | NTIS, PC A02/MF A01 - OSTI; GPO Dep. |
| System Entry Date | 2008 Feb 08 |
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