【Packing size】20 Tests/Kit
【intended use】For staining detection of sperm nuclear integrity use only
【Main components】Reagent A 50ml , Reagent B, Reagent C1 90ml, Reagent C2 13ml.
【Storage conditions and expiry date】Store at 2-8℃ for 9 months
【applicable instrument】Flowcytometer
【sample requirements】Fresh or Frozen semen。
【Test Method】
Preparation of reagent before experiment:
1)Prepare reagent C : pipetting reagent C1 78ul to reagent C2 , totally mixing by pipette
tips ,(reagent C can store at 4°C for 2 weeks )
Procedure :
1．Ejaculate is allowed to liquefy at 37 °C for 30 min，The liquefied sample adjust by adding
on reagent A to
approximate concentration,1-2 x 106 cells/ml。
2． Calibrate the flow cytometer and set the flow chamber so that the sperm flow rate
reaches 200 / SEC
Flowcytometry measurement
1.Pipetting 100 ul sperm cells ( concentration of 1-2 x 106 cells/ml) into the flow
cytometer sample tube
2.Pipetting 200ul of reagent B ( Keep in cool condition by putting sample tube on ice)
3.After timing exactly 30Sec, Pipetting 600 ul of reagent C solution and mix well
4.Putting the mixed sample into sampling chamber of Flow Cytometer and start to run the
analyzing
5.Start 3 min. timing immediately for each sample running (if the flow speed is too fast ,
the cell flow rate is more than 300 / s, the sample should be diluted with reagent A again).
6.Repeat the steps for next sample testing .
7. Washing the sample pipe after testing.
【Reference Value】
Sperm DNA fragment index (DFI) <15% : sperm DNA integrity is normal
Sperm DNA fragment index (DFI) >15% < 30% sperm DNA integrity is general
Sperm DNA fragment index (DFI) >30% sperm DNA integrity is poor
