HARVARD CMA 11 User Manual

CMA 11 55 kDa & 2MDa Microdialysis Probe User's Manual
intlet
steel shaft
membrane
Note: The 500 kDa and 2 MDa Cut Off membrane has to be use in a Push-Pull system to avoid loss of fluid out to the
surrounding tissue (ultra-filtration). Here the use of a CMA syringe pump and REGLO ICC Pump is recommended.
Instructions for CMA 11 500 kDa & 2MDa Microdialysis Probes
Fill a microsyringe with perfusion fluid and mount it in the CMA Syringe Pump. The Perfusion Fluid must be clean, at
1.
room temperature and preferably degassed.
2. Run the pump to make sure that liquid leaves the tip of the syringe cannula.
3.
Load the peristaltic pump with the "FEP Tubing Connector Peristaltic Kit". Adjust the lengths of the FEP tubing if needed for a
lower dead volume; recalculate the outlet volume if needed (1.2 µl/100 mm length)
4. Place the inlet end of the "FEP Tubing Connector Peristaltic Kit" in a beaker with Perfusion Fluid and flush the pump to fill all
tubing with perfusion fluid. Make sure there is no air bubbles in the complete tubing set.
5.
S et the pump to the required perfusion flow, usually 1 – 5 µL/min.
6. Prepare a desired length of tubing with a Tubing Adapter on both ends and connect the tubing to the inlet of the probe.
Remove the protection tube from the probe carefully. Short cannula = inlet, long cannula = outlet. Tubing Adapters and FEP
tubing should be used for all connections. To facilitate the handling of Tubing Adapters, they should be soaked in Ethanol
70% for minimum 10 minutes.
7.
Mount the probe in a Probe/Guide Clip on the CMA 130 In Vitro Stand. Put the probe membrane into a vial filled with
perfusion fluid. Connect the inlet tubing of the probe to the syringe cannula by sliding the Tubing Adapter over the cannula.
8. Flush the probe with perfusion fluid at 8-10µL/min for 4-5 min to wash out air. Knock on the shaft of the clip to help the air to
flush away. At this process the Ultra High Cut Off membrane will look as leaking but this is due to ultra-filtration of fluid
through the membrane.
9. Set the pump to the required perfusion flow, usually 1 – 5 µL/min. Make sure both pumps have the same flow rate.
10.
Connect the inlet "FEP Tubing Connector Peristaltic Kit" at the peristaltic pump to the outlet of the probe
Lift up the probe from the vial and check that the membrane doesn't ultrafiltrate or dry out; it should look filled out but
11.
not sweating. Lower the probe into the vial and control the outlet volume for the complete system.
The system with the probe is now ready for use.
12.
When changing sample vials, remember to consider the internal volume in the system (see TECHNICAL INFORMATION).
13.
This causes a delay that must be calculated when using low perfusion rates and short sampling times
outlet
shaft 0.40
0.28

TECHNICAL INFORMATION

Membrane
Material
Molecular Cut-Off
Outer Diameter
Length
Probe Shaft
Material
Diameter
Length
Internal Volume
Inlet Volume
Outlet Volume
Polyethersulfone (PES)
500kDa, 2MDa
0.28 mm
1, 2, 3 and 4 mm
Stainless-steel
0.40 mm
14 mm
0.1 µL
1 µL