Is it possible to decontaminate a vibratome (slicer) that has previously been used on fixed tissue to be used to make fresh brain slices? Has anyone tried this? I've been told that even trace amounts of fixative will ruin fresh brain slices.
Hello Josepd,
for the machine itself probably yes, but it is better to use a different set of chamber, blade and blade holder (in our model of vibratome they can be removed easily for washing and I think most fo them do). It is true that even small traces of fixative can influence the fresh tissue, when I dissect I never use the same tools that I use for fixed tissue. On the other hand, I once forgot to change the set of tools at the vibratome and cut a non fixed sample with the one we use for fixed ones. The result of the experiment was not too different from the others, but it was only a Calcium imaging experiment done the same day, I don't know if this migth have affected the result on a longer time or on an electrophisiology experiment.
Good luck
Yes, I have successfully done so, but meticulous cleaning is required to remove all trace of fixative. I would suggest also using a spray on aldehyde scavenger.
Thanks so much Elia and Donald, this is incredibly useful. Donald would you be able to recommend an aldehyde scavenger? I hadn't heard of this sort of product before.
Hey Joe,
We use the same vibratome for fixed and fresh slices (acute slices for voltammetry recordings) but have a different chamber blade holder, tools etc. I also use a tray to keep any of my fixed stuff off the bench and then wipe the bench down with normal cleaning spray just to be sure. We have had instances where people have mistakenly used the bits for fixed tissue to prepare acute slices and there doesn't seem to have been any ill effects, although in our experience voltammetry recordings are a little more robust than e-phys ones, so e-phys recordings could be more susceptible. I wash the equipment and bench down thoroughly in between use with normal detergent and lots of water. Hope all's well with you!
Hello,
I can just follow the recommendations of other colleagues.
As an electrophysiologist I am extremely fussy regarding slices preparation. Although one might be able to clean the vibratome from fixative traces one can never be sure!
Since one of the major steps to insure cell recording in slices is actually the slice quality; I would personally recommend to use a full different set of vibratome tools.
Best
I know of one company that markets commercial aldehyde scavenger. www.fronine.com.au/files/msdss/ScavengerIII%20MSDS.pdf
Having said that I think the advice given by others to use separate instruments and chamber is probably the best solution.
Yes I have successfully done this but you will have to clean it very well.
Hello Joseph,
Yes, i have done it for uncaging glutamate and laser scanning photostimulation experiments (LSPS), but a detailed cleaning is required to remove all the fixative.As Elia mentioned already don't forget to use a different set of chamber, blade and blade holder and/or wash them very well.
Good luck with your experiments.
Yes, after adequate washing (EtOH and dist-water). You vcan also replace the blade.
However, whoever warned you about the fixative (even trace amount) was right. So, that cleaning should be very thorough. I have not used such vibratom, but used surgical tools that have been used to manipulate with fixed tissue previously. Thorough washing was fine. You may have to wash a little more because the plastic container of the vibratome likely absorbes the fixative stronger than metal parts of my surgical tools.
We've used 5% HCL to clean dissection tools and a vibratome that have been used for fixed tissue and that seems to have worked. This was mainly used on the plastic casing of the vibratome, not on the specimen chamber/blade holder. Wash HCL away with water afterwards to avoid corrosion of metal parts. For tools a few minutes soaking is sufficient, glassware can sit in HCL for at least 5 min. Also, you could buy a separate chamber and blade holder like others have suggested. Also, I wouldn't switch back and forth between living and fixed tissue frequently, only use the same vibratome if you really have to. Good luck!
Wow, I'm overwhelmed by the quantity and quality of responses. Thank you all very much. And here I thought being at the tip of Africa would isolate me from the global research community.
After all that has been said, I just have to emphasize to use a dissection kit esclusively for preparing fresh brain slices. Also if you only have one vibotome don´t use it on fived material as long as you are preparing living slices. Of course after thorough cleaning and aldehyde decontamination.
Yes,we are also performing fesh and fixed tisseu slicing on the same vibratome. Considering the volume of tissue and the volume of solution in the slicing chamber, the % of fixative is going to be at best at trace levels, especially after rincing the chamber. If you are seriously concerned about an eventual effect, you can use a different chamber for each type of use.
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