Does anybody have any experience or input staining and cover-slipping a slide of interest, marking on the underside regions of interest, then removing the cover-slip and dissecting out regions of interest using LCM? Would the process likely damage DNA too much to be usable?
The standard procedure is to identify your ROI on a stained slide and then perform LCM on an unstained, unmounted slide. You can find the ROI even if not stained with phase contrast or pattern recognition. I understand you dont want to do this.
I have succesfully scraped HE mounted slides for DNA analysis (gene sequencing) for BRAF V600E mutations. You cannot LCM them as most procedures require a special coating or support slide.
HI I used PALM in the past and yes it is possible. You need to contact zeiss for the coated slides otherwise doesn't work. If you do so, then you can cut it with the laser provided by the microscope and pulse it into the eppendorf containing any sterile solution you prefer as long as is dna free (i used sterilesd pbs 100microL in the bottom of it from thermo, you shall find one DNA/RNAase free) but you need to try several times until you become proficient before you use your samples. For the staining i can't help you because my structures were visible so had no need to stain them
We do have a protocol for tissue transfer onto MMI Membrane Slides. The tissue can be stained on the membrane slides, using cresyl violet for example which is a very gentle staining. If you want to avoid staining, our software facilitates the serial section approach as suggested by Arnaud. It's only a few clicks in the software to transfer marked ROI from the stained to the unstained slide.
In case you have any further questions, please let me know.
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