Related Experiment Videos
A fast perfusion system for single cell physiology optimized for microscopes with water immersion objectives.
F Kirchhoff1, C Ohlemeyer, H Kettenmann
1Department of Neurobiology, University of Heidelberg, Federal Republic of Germany.
Pflugers Archiv : European Journal of Physiology
|April 1, 1992
Summary
This study introduces a novel perfusion system for rapid solute application under microscopy. The system enables fast, precise solution exchange, ideal for calcium imaging in tissue slices.
Area of Science:
- Microscopy and Imaging Technology
- Physiology and Neuroscience
Background:
- Accurate and rapid application of solutions is crucial for physiological studies, especially in live tissue.
- Existing perfusion methods can be slow or require complex setups, limiting experimental efficiency.
Purpose of the Study:
- To develop and validate a novel perfusion system for rapid, precise, and reproducible solute delivery to specimens under a water immersion objective.
- To optimize the system for calcium ([Ca2+]) measurements in tissue slices using fluorescence microscopy.
Main Methods:
- Construction of a five-channel pipette integrated into a water immersion objective lens holder.
- Utilizing computer-controlled magnetic valves for solution reservoir gating.
- Employing fluorescence measurements to determine solution exchange rates.
Main Results:
- The perfusion system allows for the rapid application of up to five different solutions.
- Complete solution exchange between the objective and specimen occurs within 1-2 seconds.
- The system eliminates the need for a micromanipulator, simplifying setup.
Conclusions:
- The developed perfusion system offers a significant advancement for time-sensitive experiments, particularly calcium imaging in upright microscopy.
- It provides a fast, reproducible, and precise method for solution exchange, enhancing experimental capabilities.