The Brain Slice Keeper has been designed to pre-incubate brain slices prior to transfer into recording chambers. It consists of a pair of closely fitting acrylic rings, upper and lower, between which its wedged a sheet of fine nylon netting, designed to be easily replaced when necessary.
- Slices maintained for many hours prior to transfer to recording chamber
- Hooks into purpose designed holding vessel (BSKH) or standard Buchner funnel for bubbling carbogen into ACSF
- Slices supported on a quick change nylon net
- Modular design simple to set up and maintain
In operation, the BSK1 is totally immersed in an ACSF-filled purpose-designed holding vessel (BSKH) into which a 95 % O2, 5 % CO2 gas mixture is supplied with a ceramic bubbler. The bubbles rising from the base of the BSKH saturate the ACSF and provide constant circulation of medium to the slices which rest on the nylon net. Since the lower ring of the BSK1 carries a deflecting shield, bubbles are prevented from being trapped under the slices and ensure continuous circulation of medium. It is important to adjust the flow rate of the gas mixture to the bubbler with the needle valve supplied with the BSKH. A fast bubbling rate will cause turbulence above the slices and may cause them to move and float away. The BSK1 together with the BSKH holding vessel can easily be placed into a water bath for regulating the incubating temperature as desired. Alternatively a standard Buchner funnel with 40 µm to 60 µm porosity sintered glass membrane can be used instead of the BSKH. When first placing the BSK1 into the BSKH, any large air pockets are removed from under the net by means of a teat pipette. When in place, the fluid level is maintained at least 5mm above the upper ring of the BSK1. This ensures circulation of fluid over the top and downwards towards the slices. The typical fluid volume with BSK1 with a BSKH is 220 ml and with a Buchner funnel it is 320 ml.
Brain Slice Keeper arrangement in purpose designed holder (BSKH)
Schematic arrangement of BSK1 with Buchner funnel and approximate dimensions of BSK1