Cycle description:

Underlined Valves are active Valve 4= active

None underline Valve is normal state. Valve 4= normal state like in drawing
All Valves in the pathway during process are mentioned.
Probe is deployed

1.) Move syringe block out, to create vacuum for external probe rinse and prime diluent

Sample Aspiration:

1.) Move syringe block out, to aspirate 12µl of sample, via sample syringe. No Valve is used.
2.) Retract probe while moving syringe block in, pushing 0,766 ml of diluent via external probe
connection 21, to rinse probe externally, Rinse liquid is drained directly, using the vacuum
generated before. Valve 3, 4, 5, 6 and 12 is used.
3.) Move probe to WBC bath
4.) Move syringe block in to push drained rinse liquid to waste. Valve 7 is used.
5.) Venting syringe block. Valve 8 is used.

HGB blank measurement

Rinsing of WBC bath and probe

1.) Rinse WBC bath and probe, pushing 1,53ml of diluent via external probe connection 21. Rinsed
liquid ends up in WBC bath. Valve 4, 5, 6 and 12 is used.
2.) Move syringe block out, to drain WBC bath and to prime diluent. Valve 1 is used.
3.) Move syringe block in, discharging drained liquid. Valve 7 is used

1st WBC dilution in WBC bath

1.) Move syringe block in, to push 1000µl of diluent into WBC bath via top bath connection port 4.
Valve 4, 5, and 6 is used.
2.) Move syringe block in, to push 500µl of diluent into WBC bath via probe, via Sample Syringe,
dispensing 12µl of sample into WBC bath. Valve 4 and 5 is used.

WBC bath now contains 1st WBC dilution

3.) Move syringe block to home position, pushing air into WBC bath bottom connection 1, bubble
mixing 1st dilution. Valve 1 is used.

Transfer of 25µl of 1st WBC dilution to RBC bath

1.) Move syringe block out, to create vacuum for the external probe rinse. No Valve is used.
2.) Move syringe block out, to aspirate 25µl of 1st dilution via probe. No valve is used.
3.) Retract probe and rinse externally, pushing 0.5ml of diluent. Valve 3, 4, 5, 6 and 12 is used.
4.) Move probe into RBC bath.
2nd dilution in WBC bath

1.) Move syringe block in, pushing 0.66ml of Lyse reagent into WBC bath via bath top connection
11. Valve 9 is used.

WBC bath now contains 2nd WBC dilution of 1: (125+55) (blood: (diluent + Lyse)) = 1:180

2.) Move syringe block in, rinsing probe externally over RBC bath, pushing 1ml of diluent. Valve 4,
5, 6 and 12 is used.
3.) Move syringe block in, pushing air into WBC bath bottom connection 1, bubble mixing 2nd
dilution. Valve 1 used.

RBC dilution in RBC bath

1.) Move syringe block out, to drain RBC bath. Valve 2 is used.
2.) Move syringe block in, discharging all liquid. Valve 7 is used.
3.) Move syringe block in, pushing 1,5ml of diluent into RBC bath via external probe connection21.
Valve 4, 5, 6, and 12 is used.
4.) Move syringe block in, pushing 0,5ml of diluent into RBC bath via probe, via sample syringe,
push out 25µl of 1st WBC dilution. Valve 5 and 4 is used.

RBC bath now contains RBC dilution 25µl( 1:125) (Blood:Diluent) : (2000) diluent= 1:10000
( blood:diluent)

5.) Move syringe block in, bubble mixing of RBC dilution in RBC bath. Valve 2 is used.
6.) Move syringe block in, bubble mixing 2nd WBC Dilution in WBC bath. Valve 1 is used.

Measurement

1.) Move syringe block out, to create measurement vacuum. Valve 10 is used.
2.) 1st measurement sequence CBC for 3 seconds.
3.) Move syringe block in, to generate counter pressure and perform backflush. Valve 4, 5, 6 and 12
is used.
4.) Move syringe block out, to create measurement vacuum. Valve 10 is used.
5.) 2nd measurement sequence CBC and Diff for 7 seconds.

Cleanup, preparation of next measurement

1.) Rinsing of measurements heads using cleaner. Valve 11 is used.

2.) Move syringe block out, draining of both RBC and WBC bath. Valve 1 and 2 is used.
3.) Move syringe block in, discharging all liquids and pushing diluent to WBC bath in parallel. Valve
4, 6 and 7 is used.
4.) Move syringe block in, pushing diluent to RBC bath, via simple syringe. Valve 4 and 5 is used.
5.) Retract probe while performing external probe rinse. Valve 3, 4, 5, 6 and 12 is used.
6.) Move syringe block out, priming diluent line. Goes through Valve 4.
7.) Deploy probe for next cycle.