This article details a method for collecting interstitial fluid (ISF) from the brain of an anesthetized mouse using a microdialysis probe. The process involves implanting a guide cannula and utilizing a specialized semi-permeable membrane to facilitate the collection of extracellular proteins for analysis.
Take an anesthetized mouse implanted with a guide cannula in its brain.
A dummy probe inside the cannula prevents the entry of tissue debris.
Place a collar around the mouse's neck.
Replace the dummy probe with a microdialysis probe containing a specialized semi-permeable membrane, which allows the flow of molecules with a high molecular weight.
The probe is connected to pumps through inlet and outlet ports to regulate the flow of a physiological buffer.
Tether the mouse inside a cage using the collar, allow it to recover, and initiate the flow.
The buffer enters the brain tissue through the inlet and mixes with the interstitial fluid, or ISF, in the extracellular space, which contains proteins involved in cellular communication.
The components diffuse across the membrane and are collected through the outlet in fractions.
The collected ISF, containing extracellular proteins, is ready for analysis.
Place the collar around the neck of the anesthetized guide cannula implanted animal, and remove the cap nut and dummy probe. Slowly insert the microdialysis probe through the guide cannula, and fasten the cap nut.
Then, place the mouse in a cage connected to a free moving system, and tether the mouse with the collar. After at least one hour, sequentially stop the roller pump and the syringe pump, restarting the pumps with the syringe pump set to 20% faster than the roller pump, and place the free end of the outlet tubing on a refrigerated fraction collector to collect the brain ISF. When the appropriate experimental volume has been obtained, remove the probe, and handle the mouse recovery as just demonstrated.
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