Until now, the accuracy of automated sperm analyzers was pushed to the limit when counting low density or high detritus samples (such as analyzing sperm in egg yolk extenders or homogenized tissue). Using fluorescent strobe illumination, IDENT breaks through this barrier and reduces counting errors to as little as ± 2%.
The IDENT fluorescent system is carefully engineered for seamless integration into the internal optics of the IVOS II. Only minimal adjustments are required between standard and fluorescent analysis modes. Because all analysis parameters are set and stored in software, switching to IDENT illumination may be easily accomplished with one mouse click.
Wuh the new CASA II software, setting the illumination is easy with the Interactive Illumination. Simply select the IDENT Fluorescence setup and set the Minimum Had Brightness so that sperm heads appear blue on the screen.
The strobed illumination capability of the IVOS is highly beneficial when using the IDENT fluorescent source. By limiting exposure to the UV light, the strobed illumination dramatically reduces phototoxic effects on the stained sperm. Not only does fluorescent strobe minimize bleaching of the stain but it also makes it possible to analyze fluorescently-stained, motile sperm (ideal for post-thaw motility studies).
Accurate identification of sperm is easy with use of the IDENT Stain - a specialized, DNA-specific, fluorescent dye. Since sperm heads consist mainly of condensed DNA, the dye is readily absorbed and sperm are easily visible under the fluorescent lighting. In contrast, most detritus does not contain significant amounts of DNA and therefore does not absorb enough dye to be visible under the same lighting conditions.
The two screens below show the same field viewed with and without the use of the IDENT fluorescence. The non-sperm debris virtually disappears from the analysis field.
Sample preparation and staining require only a few steps. Simply place an aliquot of semen and culture medium (for motile samples) or distilled water (for static counts) into the reaction vial and vortex. Within two minutes the sperm are stained and ready to be analyzed. The prepackaged IDENT Stain is available in kits of 200 tests.