Hello everyone,
I have recently started using the microtome device for sectioning of paraffin-embedded mice lung. While I had some success in sectioning and observing proper ribbons, some of my samples show light chatter or tears in the region of the tissue.
I have used a properly sharp blade and optimized the angles. I also made sure to use a cold paraffin block but the issue persists. Any advice to prevent/troubleshoot this would be highly appreciated.
Not all specimen blocks are created equally, even from the same processing session. Sometimes, for some blocks, you may have to change the angles (knife or clearance), the speed of sectioning, or the temperature of the blocks.
Hello Brent,
Thank you so much! I realized that a sharp blade along with its angle and rehydrating the block helped me fix it, enabling me to get proper sections. However, do you happen to know what's causing these white lines? I can't seem to get a perfect ribbon due to these. I also think that they may lead to problems when it comes to slide observation after staining.
Thank you once again!
Brent E Gowen
Those are knife marks from the blade you are using. Once the sections are stained and a coverslip is mounted, these marks tend to disappear unless the knife marks are more severe.
You should check your sections as you collect them. If you can see the marks with the naked eye, you should either move the blade to an unused area or, if this is not possible, use a new blade.
Cheers
B
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