ChemoMetec SP-100 - Manual (Page 63)

9 Technical specifications
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9 Technical specifications
Specificity The NucleoCounter SP-100 counts sperm cell nuclei stained with the DNA specific
fluorescent dye, PI.
Total Count: Counts total number of sperms (using Reagent S100, a
lysing/dilution medium)
Non-Viable Counts: Counts only the “dead” sperms (using PBS or an extender as
the dilution medium).
Sample types Mammalian semen, including semen from boars, bulls, dogs, stallions, rams, he-
goats, bucks (rabbit) etc.
Semen doses used for Artificial Insemination can also be analyzed with the
NucleoCounter SP-100
Sample consumption Total Count: The sample is mixed, lysed and diluted with Reagent S100 prior to
analysis. It is recommended that at least 50 µl of sample is used for this dilution.
Non-Viable Counts: The sample is mixed and diluted with PBS or extender prior to
analysis. It is recommended that at least 50 µl of sample is used for this dilution.
Reagent consumption It is recommended that 10 ml of reagent (Reagent S100 for a Total Count or
PBS/Extender for a Non-Viable Count) is used for dilution of a 50 µl sample of a
boar ejaculate. This corresponds to a 201-fold dilution of the sample.
When analysing a boar semen dose which contains approximately 30 million
sperm cells per ml it is recommended to add 10 ml of reagent to 1 ml of semen
dose (dilution factor 11).
The sample consumption and reagent consumption will vary from species to
species, please refer to application notes.
Sample/reagent mixture
consumption At least 100 µl of sample/reagent mixture is recommended for SP1-Cassette
loading. Approximately 60 µl of the mixture is loaded into the cassette.
Analysis volume Approximately 1 µl of the sample/reagent mixture is being analyzed in the
NucleoCounter SP-100.
Measurement range in
Semen Samples In principle all samples (ejaculates, semen does etc.) containing > 0.5 million
sperms per ml can be analyzed.
Measurement range in
sample/reagent mixture
The sample should be diluted with reagent so that the sample/reagent mixture
contains a certain amount of cells in the mixture. The total range, the
recommended range and the optimal range is:
Total Range -
Recommended Range -
Optimal Range -
50.000 to 7.000.000 sperms per ml mixture
500.000 to 7.000.000 sperms per ml mixture
750.000 to 5.000.000 sperms per ml mixture
The sample should always be diluted so that the cell density of the resulting
sample/reagent mixture is within the recommended range.
Repeatability (CV%) The repeatability is typical < 4% for a Total Count, provided that the cell
concentration of the sample/reagent mixture is in the Optimal Range (se above).
The repeatability for a Non-Viable Count is normally higher than for the Total
Count. The repeatability for a
Non-Viable Count is typical < 8%.
Dilution Factor (DF) DF in the range from 1 - 9999 can be programmed in the NucleoCounter SP-100.
DF below 11 is not recommended without first consulting ChemoMetec.
Presentation of Result The sperm cell density in the sample/reagent mixture is multiplied with the
dilution factor giving the sperm cell density in the sample. The result of the