A microperfusion chamber for study of XXXX spermatozoa.

@entity1 @entity95 @entity1319
The design of a microperfusion chamber is presented for use with spermatozoa or other cell suspensions. This chamber allows perfusion of a small number of spermatozoa during simultaneous observation of cell behavior at the microscope. The chamber is made from a flat glass capillary tube that is fitted at both ends with a filter unit containing Millipore filter discs. The entire assembly is designed to fit the stage of an inverted microscope. A population containing as few as several hundred sperm cells may be observed in the chamber during successive changes of the suspending medium as controlled by a perfusion pump. Several experiments are presented demonstrating sperm survival in the sealed chamber and the response of @entity95 and @entity1 sperm @entity1319 after the washing process. For these manipulations, the percentage of motile cells, linear swimming speed and incidence of hyperactivated @entity1319 are reported. Simple incubation in the chamber for 1 hour was not deleterious to the @entity1319 of @entity95 spermatozoa. @entity1 seminal spermatozoa showed no decline in vigorous @entity1319 after the washing procedure. Compared with in vitro capacitated spermatozoa, however, washing of @entity95 seminal spermatozoa showed a variable response. Finally, partially capacitated @entity1 spermatozoa were examined for any alteration of @entity1319 during chamber incubation with a subsequent wash. When small numbers of spermatozoa or other cell types must be manipulated, the methodology can be effectively substituted for the standard washing procedure that uses repeated centrifugation and resuspension.
