Would the temperature fluctuations from agarose embedding cause damage to the cells?
It is true that there is some temperature fluctuation during the embedding process. But what we found is that the embedding allows us to dramatically slow, and sometimes eliminate the knife’s vibrating frequency, greatly reducing the mechanical stress on the slices. In addition, we will be providing you with the tools to make this temperature fluctuation as mild and as quick as possible. Here is how we do that. a. We provide a brass block with a hole in the middle that fits the embedding tube snugly. You first place this block on ice to chill it. b. Melt the agarose (with gel point at 37 degrees Celsius), and then bring it down to around 39 to 40 degrees in a water bath. c. Pipette the agarose into the specimen tube. d. Put brass block around the tube to chill the agarose and the tissue down to the pre-embedding temperature within a few seconds.
