I am using a plastic wrap to pick up the section and transferring to a glass slide. Unfortunately, when I take off the plastic wrap some of the section attached to the plastic wrap.
Thanks for the help.
Thanks for your reply. I am actually working with sledge microtome for frozen sectioning of a whole mouse head. I am not using cryostate.
Amir, in my experience, the most effective way to manipulate sections from a freezing microtome is with a thin brush (I used to use 00).
You can collect the section from the blade with no problems at all. After collection I used to either leave sections in a vial with PBS (phosphate buffer, if I was going to work on them immediately) or in sucrose (30%) if I was going to freeze them and work on them in the future.
Mohamed, the freezing microtome has a freezing unit were the brain is placed (I also used to cut the whole brain Amir), no chamber is needed, the section defrosts has you cut them and stays in the blade, where you can collect it.
I hope this helps you.
You can put sections into the small water tank (some sort of small open box) right after the cutting and then collect the brain sections from the water directly - you need to put the glass slide under the surface of the water and then gently push the brain section and position it on the slide - no worries will not fall off.
Cheers,
Hi Mohamed,
The freezing microtome that Amir is talking about is different from a cryostat. The actual blade does not have to be in a chamber, not even frozen. The plate where you place the brain is the bit that gets frozen. With a freezing unit you can go quite down in temperature without the need for it to be in a chamber (under -50 degrees).
The MICROM HM 450 would be an example of it.
The tube you can see at the left is connected to the plate (grey metal square) and to the freezing unit (which you cannot see).
When you place the brain in the plate it gets completely frozen and you can cut it without problems by moving forwards and backwards the blade (placed in the black part).
I hope the image can help.
In my experience with PFA perfusion- and post-fixated brains, I agree with previous comments that collection with a paintbrush works very well. For the sake of efficiency, slicing and collecting all your tissue, and then working away at mounting them on pre-treated slides (e.g. gelatinized) may be a useful approach--although a steeper learning curve if less familiar with the anatomy.
A series of multiple sections can be accumulated in one container/section/well of a container (e.g. mesh baskets). The series of sections obtained, even if all mixed together can be re-ordered with ease with say 1 in 5 section series. Furthermore, prior to cutting, making an unobtrusive cut/mark somewhere in your tissue can help easily orient your sections (left/right).
Good luck!
I agree with Guillaume about the use of series and the marking to distinguish left and right hemispheres. I've used both strategies.
Amir, sorry about misspelling your name (I'm slightly mortified!). I've corrected it in my responses.
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