ChemoMetec SP-100 - Manual (Page 20)

1 Basics of the SP-100 system
2
purposes. First, the Reagent S100 is used for disruption of the plasma membranes,
rendering the nuclei of the sperm cells susceptible to staining with PI. Secondly, the
reagent is used for dilution of the semen sample. The mixture of the semen sample and
Reagent S100 is called a
lyzate mixture or sample/reagent mixture .
In ejaculates from boars, the sperm cell density is normally within the range from 100
to 1400 million cells per ml. In order to obtain an accurate and precise sperm count with
the NucleoCounter SP-100 it is recommended to dilute the boar ejaculate 201 times (50
µl of ejaculate + 10 ml of Reagent S100). This corresponds to a sperm cell density in the
range from 0,5 to 7,0 million per ml in the lyzate mixture. Approximately 1 µl of the
lyzate mixture is counted in the SP1-Cassette, corresponding to a total of 500-7000
sperm cells per analysis.
Ejaculates from each animal species have its own optimal dilution factor depending on
the typical sperm cell concentrations for the species (refer to Application Notes).
Semen doses for AI can also be analyzed with the NucleoCounter SP-100. The sperm cell
concentration in these samples is most often reduced compared to the ejaculates.
Therefore, the dilution factor is lower for these samples than for the corresponding
ejaculates. The samples should be diluted so that the final cell concentration in the
lyzate mixture is in the range from 0,5 to 7,0 million per ml. Refer to Application Notes
for the optimal dilution factor.
An appropriate dispenser (e.g. Dispensette  III, Brand, Germany) is mounted on the top
of the Reagent S100 container. A sample cup holder is also mounted on the container.
For safe handling of the reagent, the container is placed in a holder.
Figure 4. Reagent S100 container placed in a holder and equipped with a bottle-top
dispenser and a sample cup holder.
Bottle-top
Dispenser
Sample Cup
Holder
Holder for reagent
Container
Reagent S100
Container