The chamber consists of a robust acid resistant polycarbonate frame whose floor is formed by a standard 22 × 40 mm coverslip attached by vacuum grease or glue. The use of #1 or #1.5 thickness coverslips in the chamber floor enables use with high N.A. objectives on inverted microscopes. The 14.5 mm circular central working surface of the main bath accommodates 12 mm coverslips and anchors for culture work or a full rat brain slice and harp. This wide working area combined with beveled 3 mm high edges is ideally suited to upright microscopes employed in brain slice electrophysiology. This design permits low angle (down to 25°) access for multiple electrodes approaching the bath over 240° of its circumference. Perfusion input and output ports in the chamber allow direct attachment of tubing without the need for space-occupying magnets and tube holders. Standard 1.57 mm outer diameter/1.14 mm inner diameter tubing is recommended for high perfusion rates (2 - 10 ml/min) or narrow bore 1.52 mm outer diameter/0.86 mm inner diameter tubing is recommended for slower perfusion rates (0.5 - 3 ml/min). The level of the bath can be adjusted with the level of the outflow port tube which sucks from an outflow ramp. A 5.5 × 6 mm bath between the main bath and the outflow port allows placement of a variety of reference electrodes. The eye shaped main bath is optimized for laminar flow perfusion.
The chamber can be fitted to existing chamber holders from Warner (series 20) or Bioscience Tools (PLD-A) or can sold together with a 3 mm thick 80 mm diameter circular polycarbonate holder fitting Olympus stages (other diameter on request). A 10 mm high ring around the holder’s outer circumference guards against flooding.
This chamber was developed and tested in cooperation with the Interdisciplinary Center for Neurosciences (IZN), Heidelberg.