dear following steps used:

1. Turn on the Z1 by flipping the switch on the top left of the instrument to the “ON” position

2. Open the door to the instrument and carefully lower the black platform to the bottom position

• The platform has a safety latch that must be pressed to allow the platform to move

3. Remove the Z1 cup containing the blue Coulter Clenz solution

• Save this cup and the Coulter Clenz solution for when you are finished

• Periodically, this solution should be replaced with fresh Coulter Clenz solution from the jug next to the sink

4. Fill the “FLUSH” Z1 cup with 10 ml Isoton II Diluent

• The pump on the Z-Pak fill dispenser automatically dispenses 10 ml

• Be sure to remove the screw cap on the spigot before trying to dispense

5. Put the “FLUSH” Z1 cup on the black platform of the instrument and raise the platform to the top position

• The platform will stop before the glass probe hits the bottom of the Z1 cup, so do not worry about smashing the probe

6. On the instrument keypad, press “Function” and scroll down until “Prime Aperture” is displayed

7. Press “Start” twice to begin priming the aperture

• Aperture priming replaces the fill solution in the probe with fresh Isoton II Diluent

• The priming procedure takes ~5 min

8. While the instrument is priming, prepare another Z1 cup with 10 ml Isoton II Diluent and 50 μl of the cell suspension to be counted

• This 200-fold dilution is coded into the instrument and is factored into the cell concentrations shown

• If another dilution is used, then the concentrations returned will not be accurate

9. After the priming has completed, lower the black platform of the instrument to the bottom position and discard the remaining Isoton II Diluent from the “FLUSH” Z1 cup

• If measuring more than one cell suspension, refill the “FLUSH” Z1 cup with fresh Isoton II Diluent (the priming procedure leaves some residual Coulter Clenz in the “FLUSH” Z1 cup)

10. Put the Z1 cup containing the diluted cell suspension on the black platform of the instrument and raise the platform to the top position

• To ensure that the cells have not settled, invert the cup once or twice before removing the cap and putting the cup on the platform

11. On the instrument keypad, press “Output” and then “Start” to begin counting

• Be sure to inspect the particle counts when the instrument is counting

• The most-accurate readings occur when the number of counted particles is between 500–5000, which corresponds to cell concentrations between 2 × 105 – 2 × 106 per ml (each count draws 0.5 ml of the Isoton solution)

• If your cell concentration is much lower or higher, the cell suspension should be diluted or concentrated and then recounted

• The counting procedure should take ~15 sec

12. The concentration shown on the instrument is the concentration of the original cell suspension

13. If measuring additional cell samples, lower the black platform of the instrument about halfway to expose

the probe, rinse the probe with a squirt bottle, and remove the Z1 cup containing the counted cell suspension

14. Put the “FLUSH” Z1 cup (containing fresh Isoton II Diluent) on the black platform and raise the platform to the top position

15. On the instrument keypad, press “Function” and scroll down until “Flush Aperture” is displayed

16. Press “Start” once to begin flushing the aperture

• Aperture flushing draws a volume of clean Isoton II Diluent through the aperture of the probe

• The flushing procedure should take ~15 sec

17. Repeat steps 10–16 for each additional cell suspension to be counted

• Once the “FLUSH” Z1 cup has been replaced with fresh Isoton II Diluent after the priming step, this solution can be reused for all subsequent aperture flushes without replacement of the solution.

18. When finished for the day, Prime the Aperture (complete cycle) using a non-lytic cleaner (e.g. Beckman Coulter CLENZ) and leave the cleaning agent around the aperture overnight. Ensure that the

Aperature and the electrode are submerged in the cleaning agent before setting the Power Switch to O (off).

19. Used Z1 cups should be rinsed with DI water, sprayed with 70% EtOH, and left to dry for the next user

20. The Counter should be disinfected periodically following Appendix 7 in the User’s Manual.

Thank you

I followed basically what you described, I just have an additional question.

How do you convert your counts to cells/mL 

I have counts of 500  what is the # of cells/mL

Thanks again