Use of Coulter Counter

 

Materials

Cell suspension

Isoton

Coulter Counter (CC) vials

 

Safety

1.      Wear disposable latex gloves at all times

2.      Wear safety glasses at all times.

 

Machine Set-Up (accomplished by Instructor)

1.      To start-up, turn machine on.  Replace Coulter Clenz solution with Isoton.  Run “fill system” protocol.

2.      To shut down, set Coulter Clenz solution on platform.  Run a standard measurement.  Turn off machine. 

 

Set-Up/Preparation

1.      Confirm that Coulter Counter is turned on.  Do not adjust either of knobs on front panel.

2.      Prepare samples.

·        CC vial with 20 ml Isoton (blank)

·        Cell samples prepared by diluting 500 mL cells in 9.5 mL Isoton (or, 1 mL cells in 9 mL Isoton).

NOTE:  You may need to adjust the volume on the Isoton dispensers.  Unscrew red knob to loosen;  change volume;  screw knob to tighten.  The red cap on the end of the dispenser must be removed for Isoton to flow.

 

Sample Placement

1.      See instructions on Z1 or Z2 Routine Counting.

2.      Gently push black lever in and pull down on platform.

3.      Gently mix the solution vial two or three times by inversion.

4.      Place CC vial on platform.

5.      Gently push black lever in and push up on platform.  (Light should come on.)

6.      Check black box in upper right corner of Coulter Counter to ensure that aperture hole is clear and not blocked with debris.

 

General Operation

1.      Run Isoton blank to ensure that particle count is low (<50).

2.      Gently mix the sample 2-3 times by inversion before placing it on the stand.

3.      Run cell samples as needed.  See charts on following pages for specific steps.  Samples can be run once or in triplicate. 

4.      Rinse the aperture with DI water between samples to avoid cross contamination.  To gain access to the aperture, open the front panel.  Use a squirt bottle of DI water to rinse the aperture and mixer blade.  Be careful not to touch/push/bend the aperture or mixer blade.  Use an empty CC vial to catch drips of water.

5.      If aperture is clogged, hit “Unblock” button and/or use pink brush to wipe aperture clean.

6.      After every 5 samples, run Isoton blank to make sure background count is low (<50).

7.      Ensure that samples do not “run dry” of fluid.  (As general rule, vials should contain 7 mL - 20 mL of fluid.)

8.      At end of operation, rinse the aperture with DI water. Run Isoton blank to make sure background count is low (<50).  Leave aperture in Isoton blank for next user.  Do NOT leave the aperature exposed to air or in a sample with cells;  this may damage the aperature.

9.      Clean area around Coulter Counter.

10.  Decontaminate cell samples and dispose of biohazardous waste appropriately.