Cleaner Line.

1. Cleaner reagent is used to:

• Deproteinize the counting apertures at each analysis cycle avoiding clogging.

• Deproteinize the counting chambers and the counting apertures, when shut down selected.

2. Cleaner Line.

3. Cleaner line is composed specifically with 1 valves 2/2.

V11: <OFF> Cleaner line is closed.

<ON> Cleaner line is opened.

1
 4. Cleaner prime.

NOTE: Because there is no dedicated syringe for cleaner, system will use valves #8 and #10 and waste syringe to
prime the cleaner solution.

• Valves #11, #8, #10 <ON>.

• Syringe motor moves pistons down.
• Cleaner is aspirated inside the waste syringe.

2
 5. Cleaner into apertures (backflush)

• Cleaner is already primed at the back of the apertures.

• Syringe motor moves pistons up.
• Valves #4, #12 <ON>
• Thanks to diluent line, cleaner is pushed through apertures.

NOTE: Typically, this case is used at the end of each analysis cycle to perform a backflush into the apertures.

3
6. Cleaner into counting chambers.

6.1 Cleaner Prime.

• Valves #11, #8, #10 <ON>.

• Syringe motor moves pistons down.
• Cleaner is aspirated inside the waste syringe.

4
 6.2 WBC bath filled with Cleaner

• Syringe motor moves pistons up.

• Valves #1 <ON>

5
 6.3 Cleaner Prime.

• Valves #11, #8, #10 <ON>.

• Syringe motor moves pistons down.
• Cleaner is aspirated inside the waste syringe.

6
 6.4 RBC bath filled with Cleaner

• Syringe motor moves pistons up.

• Valves #2 is ON

7
 Fluidic Diluent Line.
1. Diluent reagent is used to:

• Perform the dilutions WBC and RBC.

• Make the sheath in the optic flow cell.
• Wash the needle internally and externally.
• Rinse the needle externally.
• Be present at the back of the apertures during the counting.

2. General Diluent Line.

3. Diluent line is composed with 6 valves 3/3.

V4: Aspiration of the diluent into the syringe. V18: Dispense diluent to V12.
Dispense diluent to V5. Dispense diluent to RBC bath

V5: Dispense diluent to V6. V12: Dispense diluent to rinse head.

Dispense diluent to internal needle. Dispense diluent to V15

V6: Dispense diluent to V18. V15: Dispense diluent to apertures.

Dispense diluent to WBC bath Dispense diluent to flow cell (sheath)

1
4. Diluent aspiration in diluent syringe.

• All valves in normal state, as shown in the drawing.

• Syringe motor moves pistons down.
• Due to the normal state of valve #4, diluent is aspirated inside the diluent syringe.

2
 5. Diluent to the rinse head.

• Valve #4 is ON.
• Syringe motor moves pistons up.
• Diluent is pushed through V4/V5/V6/V18/V12/Rinse head.

NOTE: Typically, this case is used to wash and to rinse the needle externally.

Needle washing: (1) The diluent is pushed to the rinse head (2) at the same time vacuum is applied at the opposite
side of the rinse head (3) the needle moves from down to up.

Needle rinsing: (1) The diluent is pushed to the rinse head without vacuum applied on the opposite side.
 Diluent fall along the needle and rinse it

3
 6. Diluent to the internal of the needle.

• Valve #4 ON
• Valve #5 ON
• Syringe motor moves pistons up.
• Diluent is pushed through V4/V5/Sampling syringe/Internal needle.

NOTE: Typically, this case is used to dispense the blood into the counting chambers, to perform 1/3 of the WBC
and RBC dilution and to rinse the needle internally.

4
 7. Diluent to the WBC bath.

• Valve #4 ON
• Valve #6 ON
• Syringe motor moves pistons up.
• Diluent is pushed through V4/V5/V6/heating system/WBC bath.

NOTE: Typically, this case is used to perform 2/3 of the WBC dilution.

5
 8. Diluent to the RBC bath.

• Valve #4 ON
• Valve #18 ON
• Syringe motor moves pistons up.
• Diluent is pushed through V4/V5/V6/V18/RBC bath.

NOTE: Typically, this case is used to perform 2/3 of the RBC dilution.

6
 9. Diluent to flow cell.

• Valve #4 ON
• Valve #12 ON
• Valve #15 ON
• Syringe motor moves pistons up.
• Diluent is pushed through V4/V5/V6/V18/V12/V15/Flow cell.

NOTE: Typically, this case is used to make the diluent sheath in the flow cell during optic measurement.

7
 10. Diluent to the apertures.

• Valve #4 ON
• Valve #12 ON
• Syringe motor moves pistons up.
• Diluent is pushed through V4/V5/V6/V18/V12/V15/apertures.

NOTE: Typically, this case is used to supply the fluidic located at the back of the apertures with diluent.

8
 Lyse Line.
1. Lyse reagent is used to:

• Release the hemoglobin, exploding red blood cells.

• Get in shape the white blood cells to facilitate their discrimination during optical acquisition.
• Push the sample into the flow cell for optic measurement.

Lyse Line.

2. Lyse line is composed with 2 valves.

V9: <OFF> Aspiration of lyse into the lyse syringe.

<ON> Dispense lyse to V14.

V14: <OFF> Dispense lyse to flow cell and discharge tubing (33B).
<ON> Dispense lyse to the heating system and then to the WBC counting chamber.

1
3. Lyse aspiration in Lyse syringe.

• All valves in normal state, as shown in the drawing.

• Syringe motor moves pistons down.
• Lyse is aspirated inside the lyse syringe.

2
 4. Lyse to flow cell

• Valve #9 is ON.
• Syringe motor moves pistons up.
• Lyse is pushed through V9/V14/flow cell & discharge tubing (33B).

NOTE: Typically, this case is used to push the sample into the nozzle of the flow cell.
At this occasion and due to fluidic construction, the portion of the lyse pushed into discharge tubing returns in
tubing 3A.

3
 5. Lyse to WBC bath.

• Valve #9 ON
• Valve #14 ON
• Syringe motor moves pistons up.
• Lyse is pushed through V9/V14/Heating system/WBC bath.

NOTE: Typically, this case is used to dispense the lyse into WBC bath

4
 MYTHIC 60 ANALYSE CYCLE - FLUIDIC SEQUENCES DESCRIPTION

1. SAMPLE ASPIRATION AND SAMPLING PROBE CLEANING

STEP ACTION COMMENT

1 HGB led on HGB Green led located in WBC counting chamber is switched on
2 Read HGB Blank HGB led measure through diluent for HGB reference
3 Syringe DOWN Vacuum tank draining
4
Syringe DOWN Sample aspiration in the sampling probe
Sampling probe UP + Syringe UP +
5 Sampling probe outside rinsing with syringe and pump
Pump ON
Sampling module moves to home
6 Sampling module and sampling probe in home position
position
Sequence duration: 5 secs, cycle duration: 5 secs

2. SAMPLING PROBE MOVES TO WBC COUNTING CHAMBER AND WASTE CHAMBERS DRAINING

STEP ACTION COMMENT

1 Syringe UP Waste chambers draining
Sampling module moves to WBC
2 Sampling probe located above the WBC counting chamber
counting chamber
3 Sampling probe DOWN Sampling probe moves to rinsing position in WBC counting chamber
4 Syringe UP Sampling probe outside rinsing with syringe in WBC counting chamber
Sequence duration: 2 secs, cycle duration: 7secs

3. WBC COUNTING CHAMBER DRAINING AND SAMPLING PROBE POSITIONING

STEP ACTION COMMENT

1 Syringe DOWN + Pump ON WBC counting chamber draining + Transfer tubing draining
2 Sampling probe DOWN Sampling probe moves to dilution position in WBC counting bath
3 Syringe UP Waste chambers draining
Sequence duration: 3.25 secs, cycle duration: 10.25 secs

4. FIRST DILUTION AND BUBBLING IN WBC COUNTING CHAMBER

STEP ACTION COMMENT

Diluent (by entry on WBC counting chamber) in WBC counting
1 Syringe UP
chamber for first dilution
Sample + Diluent (through the sampling probe) dispense in WBC
2 Syringe UP
counting chamber for dilution
3 Syringe UP First sample and diluent bubbling in WBC counting chamber
Sequence duration: 5.25 secs, cycle duration: 15.5 secs

1
 5. FIRST DILUTION ASPIRATION, SAMPLING PROBE CLEANING

STEP ACTION COMMENT

RBC counting chamber partial draining and diluent removal on WBC
1 Syringe DOWN
counting chamber entry
Syringe DOWN and Sampling probe
2 First dilution aspiration in sampling probe
UP
Sampling probe UP + Syringe UP + Sampling probe moves to home position and sampling probe outside
3
Pump ON rinsing
Sampling module moves to RBC
4 Sampling module in RBC dilution position
counting chamber
5 Sampling probe DOWN Sampling probe moves to rinsing position in RBC counting chamber
Sequence duration: 5.25 secs, cycle duration: 20.75 secs

6. LYSE IN WBC COUNTING CHAMBER AND BUBBLING

STEP ACTION COMMENT

Lyse dispense in WBC counting chamber + Sampling probe outside
2 Syringe UP
rinsing in RBC counting chamber
3 Syringe UP End of lyse dispense in WBC counting chamber
4 Syringe UP Second bubbling in WBC counting chamber
Sequence duration: 2.5 secs, cycle duration: 23.25 secs

7. RBC COUNTING CHAMBER DRAINING

STEP ACTION COMMENT

RBC counting chamber draining and lyse removal on WBC counting
1 Syringe DOWN + Pump ON
chamber
2 Sampling probe DOWN Sampling probe moves to dilution position in RBC counting chamber
3 Syringe UP Waste chambers draining
Sequence duration: 2.25 secs, cycle duration: 25.5 secs

8. SECOND DILUTION AND BUBBLING IN RBC COUNTING CHAMBER

STEP ACTION COMMENT

Diluent (by entry on RBC counting chamber) in RBC counting chamber
1 Syringe UP
for RBC dilution
First dilution sample + Diluent (through the sampling probe) dispense
2 Syringe UP
in RBC counting chamber for dilution
3 Measurement current ON
4 Optical LED ON
Third bubbling in WBC counting chamber + Sampling probe in home
5 Syringe UP + Sampling probe UP
position

6 Syringe UP Bubbling in RBC counting chamber + counting heads rinsing

Sequence duration: 7 secs, cycle duration: 32.5 secs

2
 9. FIRST COUNTING

STEP ACTION COMMENT

Waste chambers at atmosphere Pressure control to “0” in waste chambers before vacuum creation for
1
pressure counting
WBC dilution transferred to the optical bench and diluent removal on
2 Syringe DOWN
RBC counting chamber
First and second dilutions (present in counting chambers) aspiration
3 Syringe DOWN
through WBC and RBC counting heads
4 Wait 500ms Resistive counting vacuum stabilization
5 Check pressure Pressure control before resistive counting
Cells counting acquisition during aspiration through WBC and RBC
6 Start first RES counting
counting heads
7 Stop first RES counting
Sequence duration: 8 secs, cycle duration: 40.5 secs

10. BACK FLUSH IN COUNTING HEADS AND SECOND COUNTING

STEP ACTION COMMENT

Back flushes with diluent in counting heads to make sure potential
1 Syringe UP
clogs did not happen during first counting sequence
First and second dilutions (present in baths) aspiration through WBC
2 Syringe DOWN
and RBC counting heads
Syringe UP and start optical WBC
3 Injection of transferred sample in optical bench, WBC differentiation
differentiation
Cells counting acquisition during aspiration through WBC and RBC
4 Start second RES counting
counting heads
5 Stop second RES counting
Sequence duration: 10.25 secs, cycle duration: 50.75 secs

11. COUNTING HEADS CLEANING

STEP ACTION COMMENT

1 Counting Heads cleaning Diluent behind counting heads is replaced by cleaner
2 Stop Optical WBC differentiation
3 Measurement current OFF
4 Optical LED OFF
Sequence duration: 1.75 secs, cycle duration: 52.5 secs

12. WBC COUNTING CHAMBER DRAINING AND FILLING

STEP ACTION COMMENT

Syringe DOWN + Pump ON + WBC counting chamber and transfer tubing draining, sampling probe
1
Sampling probe DOWN moves to rinsing position in RBC counting chamber
2 Syringe UP WBC counting chamber filling with diluent
Back flushes with cleaner for cleaning counting heads after counting
3 Syringe UP
sequences
Sequence duration: 2.5 secs, cycle duration: 55 secs

3
 13. RBC COUNTING CHAMBER DRAINING AND FILLING

STEP ACTION COMMENT

1 Syringe DOWN + Pump ON RBC counting chamber draining and optical transfer priming
Diluent (through Sampling probe outside) dispense in RBC counting
2 Syringe UP
chamber + waste chambers draining
3 Sampling probe UP Sampling probe moves to home position
4 Pump ON Sampling probe drying through rinsing head
5 Syringe UP Diluent dispense in rinsing head to rinse the Sampling probe
Sequence duration: 2.5 secs, cycle duration: 57.5 secs

14. SAMPLING MODULE MOVES TO WBC COUNTING CHAMBER AND SAMPLING PROBE
INITIALIZATION FOR NEXT SAMPLE

STEP ACTION COMMENT

Sampling module moves to home
1 Sampling module in sampling position
position
2 Syringe UP Syringe moves to home position
3 Sampling probe DOWN Sampling probe in sampling position, ready for next sampling
Waste chambers to atmosphere
4
pressure
5 End of analytic cycle
Sequence duration: 2.5 secs, cycle duration: 60 secs

4
Sampling

All valves in normal state, pistons move down.

When sampling piston moves down it allows the blood aspiration inside the needle
RBC bubbling

1. Valve #17 and #8 are switched on, pistons move down.

 Air is aspirated in waste syringe.
2. Valve #2 is switched on and piston move up.
 Air is pushed in RBC chamber making the bubbling.
WBC bubbling

1. Valve #17 and #8 are switched on, pistons move down.

 Air is aspirated in waste syringe.
2. Valve #1 is switched on and piston move up.
 Air is pushed in WBC chamber making the bubbling.
 Fluidic Waste Line.

1. Waste line is used to:

• Drain WBC counting chamber.

• Drain RBC counting chamber.
• Drain optical flowcell during measurement.
• Drain rinse head during needle cleaning.
• Drain waste syringe.

2. General Waste Line.

3. Waste line is composed with 4 valves.

V1: Open WBC bath for draining V3: Drain rinse head during needle cleaning

V2: Open RBC bath for draining V7: Drain waste syringe, when pistons move
up

1
 4. Drain WBC counting chamber.

• Valve #1 switched on.

• Syringe motor moves pistons down.

Thanks to the vacuum created in waste syringes when pistons go down and valve #1 opened, the liquid present in
WBC chamber will be drained to the waste syringes.

2
 5. Drain RBC counting chamber.

• Valve #2 switched on.

• Syringe motor moves pistons down.

Thanks to the vacuum created in waste syringes when pistons go down and valve #2 opened, the liquid present in
RBC chamber will be drained to the waste syringes.

3
6. Drain waste syringe

• Valve #7 switched on.

• Syringe motor moves pistons up.

Thanks to the pistons moving up and Valve#7 opened, the liquid present in waste syringe is evacuated to
the waste container.

4
 7. Drain optical flowcell during measurement.

During optic measurement diluent and sampling dilution are pushed through the nozzle and naturally evacuated
to the waste container.

5
 8. Drain rinse head during needle cleaning.

• Valve #3 in normal state.

• Pump is ON.

Thanks to the pump ON and valve #3 in normal state, liquid is aspirated from rinse head and evacuated to the
waste container.