ChemoMetec SP-100 - Manual (Page 19)

1 Basics of the SP-100 system
1
1 Basics of the SP-100 system
This section describes the basic principles of the NucleoCounter SP-100 system. In the
section the attention is primarily paid to performing a
Total Count. With respect to
performing a Non-Viable Count, please refer to “ Viability Testing with NucleoCounter
SP-100, Addendum to User’ s Guide (part no. 991-0104).
1.1 NucleoCounter SP-100 system
The NucleoCounter SP-100 system is based on staining the DNA from mammalian sperm
cells using a fluorescent dye, propidium iodide (PI). When PI is added to a suspension of
chemically pre-treated sperm cells, the dye will specifically bind to the nuclei of the
cells. If the cell suspension subsequently is illuminated with green light, the dye bound
to the cell nuclei will emit red fluorescense light. When exposing an image using
microscopic technique each nucleus can be observed as dots on a dark background.
The NucleoCounter SP-100 system is comprised of the NucleoCounter SP-100 instrument,
a disposable SP1-Cassette and Reagent S100, which is a lysis/dilution buffer.
Figure 3
shows the system and the accessories and additional equipment needed to perform a
Total Count.
Figure 3.... NucleoCounter SP-100 system with accessories and equipment needed to
perform a Total Count on a Semen Ejaculate.
1.1.1 Lysis/dilution buffer - Reagent S100
A lysis/dilution buffer, Reagent S100 ( Figure 4 ), is always added to the semen sample
prior to performing a Total Count. The addition of Reagent S100 buffer has two
Sample Cup Sample
Sample Cup
Holder
Bottle-top Dispenser
Reagent S100
Container
Holder for
Reagent Container
Pipette SP1-Cassette
NucleoCounter SP-100