Anyone had experience in Laser capture micro dissection of H&E stained FFPE sections mounted on regular glass?
I found that the cutting is OK just the tissues won't be picked up. Only leaving the cutting line on the Cap membrane.
I run into this issue often due to humidity and static in the air. Depending on the model of LCM you use, the dissected tissue will either fall into the tube or will be picked up by a moving cap. If you are having trouble with your tissue and membrane sticking to the slide, place a dehumidifier in the room and keep it running. You can encourage the region to become unstuck by using the laser in manual control and just tapping on the region with the laser a few times. If that doesn't work, you may need to dissect smaller areas as these have less of a problem coming off.
I wasn't entirely sure whether you meant regular glass without a membrane, or membrane slides. You absolutely must use membrane slides when using LCM on H&E FFPE sections. Otherwise the tissue will not come off the glass at all.
It depends on which system you are using for the LCM. I know that it is possible to perform LCM on regular glass slides using the Zeiss Palm Microbeam system however i am unsure whether it's possible with other systems. But even with the Zeiss, using membrane slides is best in my experience.
I worked on Arcturus XT LCM, and mebrane slide was also my best experience with FFPE section.
The MMI CellCut Laser Microdissection system is using a unique Cap lift tool to isolate the dissected cells from the membrane. The big advange is that it does neither depend on electrostatics in the air and humidity, nor on the size or shape of the dissected area.
Please feel free to visit the website and watch the short video how this works:
https://www.molecular-machines.com/products/cellcut
Hello,
I actually have a problem with this device, namely the tissue samples don't attach to the cap. According to the protocol a glass slide should be underneath the membrane during the LCM, and the cells fall on the glass slide instead of attaching to the cap. Have you got any ideas what could be the problem? Maybe the isolation cap is old and the glue got dry? And is there a way to check if there are cells on the cap actually? Thank you very much in advance!
Dear László,
Are you using the MMI system?
If yes, there are two possible solutions.
With the MMI system it is very easy to check if your cells are on the cap. Just move to a free spot without samples (either on the same slide or on an empty glass slide), move the cap down and focus on the cap surface.
In case you have any further questions, please let me know.
Kind regards,
Heidi
- Badges
- Science topic
- Similar topics
- Biological Science
- Histology
- Sectioning