Using the Coulter Counter

 

June 1999 Revision

 

Materials

Qty

Location

Order Info

Cuvettes

 

B-013

Coulter Accuvetters 200

Diluent

 

B-013

Coulter IsotonII PN 8546720

 

 

 

 

 

 

Usual sample dilution : 0.2 ml sample in 9.8 ml diluent

 

Typical settings:            Lower threshold:           10

                                    Upper threshold:           99.9 (out)

                                    Attenuation:                  Tissue culture cells        16

                                                                        Fresh Lymphocytes      4

                                    Manometer:                  500 ul

                                    Current :                       100

 

  1. Turn on machines.
  2. Allow vacuum to develop (~2 min).
  3. Put blank sample in place.
  4. Turn top dial to "Reset" (vertical alignment).
  5. Adjust attenuation so the majority of peaks lie in the middle of third of screen.
  6. When window lights up, turn top dial to "Count: (horizontal alignment).
  7. When background counts are less than ~100, turn lower dial to "Fill", count to three then turn dial to "Close". This releases any residual vacuum so that air is not drawn into the electrode.
  8. Remove blank and put diluted sample in place.
  9. Turn top dial to "Reset".
  10. When window lights up, turn top dial to "Count: (horizontal alignment).
  11. Cells/ml in original suspension = number of counts x 2 (for 500 ul) x 50 (200 ul in 10 ml)
  12. Run Coulter cleanse through before shutting down the machine.
  13. NOTE: Make sure the top and lower dials are in the Horizontal position before leaving the machine or else it will suck the Isoton dry from the reservoir and it may even aspirate the mercury :-(