P064: RECYCLING DISCARDED ANESTHESIA AND OTHER EQUIPMENT IN A PATCH CLAMP SETUP FOR ELECTROPHYSIOLOGICAL RECORDINGS IN NEURONAL CELLS
Maksym Doroshenko, MD, MS; Danielle Horn; Dennis Patin; Roy Levitt; Konstantinos Sarantopoulos; University of Miami / Jackson Memorial Hospital
Introduction: Conducting basic science research, and in particular electrophysiological research, can be very challenging and costly these days. Preparing a functional patch clamp setup requires not only basic equipment, such as an inverted microscope, amplifier, digitizer and computer with software, but the manufacturing of a functional and low-noise perfusion system, recording bath chamber and suction for draining the solutions used in the experiments. Some of the components of the latter parts can be easily available in the operating room setting or the procedure rooms in the pain clinics. These include clean, non-contaminated material such as IV and arterial line tubing, stopcocks, manifolds and syringes. Other parts may not be available but can be purchased at very low cost.
Methods: We have hereby presented the use of recycled anesthesia equipment, as well as other gear used in hospitals for various purposes, but utilized in completing a functional patch clamp set up for electrophysiological recordings.
The used anesthesia equipment includes:
- Unused syringes (clean, non contaminated) obtained from procedure trays, and utilized instead of being discarded.
- Three-way stop-cocks, obtained from lines that were not used.
- Plastic IV or arterial line tubing (clean, uncontaminated) obtained from opened packages, but not used.
- Iodine applicator sponges and gauzes were used to clean the recording chamber after experiments.
- Removable mounting putty and dental or museum wax (purchased from pharmacies or other stores) was used to secure the recording chamber and stage in place.
- Damage-free hanging strips were used to secure tubing and cables in place.
- Aluminum foil was used to cover the Faraday cage for improved electromagnetic isolation
Suction system is necessary to allow drainage of the perfusion solutions from the recording chamber. However, not all laboratory rooms are equipped with a negative pressure vacuum system in the wall. On the other hand, portable vacuum pumps can generate vibrations and electrical noise that is unacceptable for electrophysiological recordings.
We used a simple and inexpensive aquarium air flow pump, that was modified into a vacuum suction pump after we reversed its internal valve. This was positioned onto a few plastic gloves to absorb any vibrations. This pump costs less than $10, and is essentially noiseless while it does not generate any electrical interference, but strong enough to provide adequate suction.
Finally, small Cuban coffee cups, offered for free in most local coffee shops were used to place solutions or to cover vials containing solutions.
Conclusions: Utilizing above parts made possible the completion of a functional patch clamp set up, saving money that would have otherwise been spent in purchasing specialized tubing or establishing a costly perfusion and vacuum suction system. This set-up has already being used in obtaining patch clamp recordings from neuronal cells in the current clamp and voltage clamp configurations.