Ready-to-use slides pre-stained with standard dyes for time and cost-saving differential diagnostic microscopy.
Spermatozoal staining test principle.
The stain is composed of the two dyes new methylene blue N and cresyl violet acetate in chemically pure form.
Constant mixing ratio and even distribution ensure reliably good staining quality.
Reaction principle analogous to that of the panchromatic (panoptic) stain.
The amount of liquefied ejaculate needed is dependent on the spermatozoa concentration: 5 µl (spermatozoa concentration > 50 mill/ml) 10 µl (spermatozoa concentration >10–50 mill/ml) 15 µl (spermatozoa concentration < 10 mill/ml) Apply a drop of the liquefied ejaculate to the middle of a cover slip and place on the pre-stained area of the slide in such a way that the ejaculate spreads evenly in all directions.
To ensure even distribution of the sample it is useful to press on the middle of the cover slip with a pencil.
After 30 minutes a weak staining is attainable, however the preparation should be left to stand at room temperature for about 2 hours until the majority of the spermatozoa have become immobile and well stained. Then differentiation into normal and pathological forms is undertaken.
The preparation is stable for at least 4 hours at room temperature. If microscopic differentiation cannot be performed until after 4 to 24 hours the slide should be refrigerated immediately after preparation.
After 20–24 hours stored in a refrigerator at 4 °C the spermatozoa are still well stained. Fixation using Eukitt® is not advisable due to the subsequent destruction of the spermatozoa.
Evaluation is performed at a magnification of x800 to x1000 with oil immersion. Usually 200–500 spermatozoa per preparation are evaluated and documented in percent according to the criteria for normal and pathological forms.
Characterisation of the spermatozoa
The spermatozoa stain very well. The head (b), the neck / midpiece (c) and the tail (d) can all be clearly identified.
The nucleus lying within the head stains dark purple. The plasma appears orange to pale violet depending on the focus. The mid-piece also appears pale violet if it is pathologically enlarged. The tail is only lightly stained but is clearly visible.