Experiments on the use of a microfrozen sectioning machine and the manipulation of brain tissue sections in large (small) mice
Experiments on the use of a microfrozen sectioning machine and the manipulation of brain tissue sections in large (small) mice
Frozen slicers can be used to: (2) cut thin, uniform slices of tissue; (2) support the tissue with hard paraffin or other substances, and automatically advance it the desired distance forward (in the direction of the knife) for each cut by means of a borrowed section thicknesser, which has a gradient of usually 1 micrometer.
Operation method
microtome
Principle
When paraffin-embedded tissue is cut, a slice strip is made for multiple slices due to adhesion to the wax edge of the previous slice.
Materials and Instruments
Mouse Brain Move 1. Install the cutter and move the safety lever. Adjust the temperature of the slicer to -20 ℃ ((right hand side up and down two buttons to adjust, up to adjust the temperature, down, reduce the temperature). 2. Remove the brain tissue from the -80 ℃ refrigerator and place it in the slicer or -20 ℃ refrigerator for about 1 hour in order to equalize the tissue temperature, which is too cold and may cause the brain slices to rupture during slicing. 3. Secure the tissue to the carrier tray (Blocker) with embedding agent (glue instead), making sure to place the brain in the right position. 4. Back off the specimen holder, secure the tray, and adjust the rocker so that it is correctly positioned from side to side. 7. When the time is right, apply a few brain slices to determine whether the top, bottom, left and right sides are correct, and to find out the approximate position in comparison with the atlas. Note: Place your hands on both ends of the slide and press down gently on one end against the end of the slicer blade. 8. Locate the nuclei to be found against the atlas and leave typical brain slices with SON, PVN, and hippocampus (CA1, CA2, CA3) in each and mark them. Caveat When slicing, the cut slice passes smoothly through the channel between the knife guards in the first instance and lies flat on the iron plate of the knife holder. At this point, the anti-curl plate can be lifted, a slide taken and attached. Common Problems How does the optic bundle change during slicing? And draw the shape of the optic tract when SON, PVN and hippocampus appear. For more product details, please visit Aladdin Scientific website.
Surgical Instruments Microscopic Frozen Section Machine Embedding Agent
Mouse Stereogram
5. Open the safety lever. Advance the tray to the proper position (not too close to avoid accidental damage to the knife, and not too far away).
6. Adjust the knife spacing (section thickness and trim thickness), crank the handwheel and start slicing. At the beginning, you can choose a larger knife distance (trim thickness, 10-500 um), to a certain position when the use of adjusted to the desired final section thickness (section thickness, 1-20 um).
After determining the SON, calculate the number of cuts left according to the atlas, cut off a few cuts, patch, stain with Evans blue for about a minute, pour off the staining solution, and observe under the microscope.
9. Store the brain slices in the refrigerator at -20 ℃ or -80 ℃ for later use.
10. Clean the slicer and recover.
Source "Neurobiology Laboratory Handout" edited by Du Jizeng and Chen Xuequn.