I cut thin sections (14 um) of fixed frozen brain and spinal cord tissues and mount on Superfrost Plus slides. While I'm sliding them, I let them dry for 10-15 mins at RT and then I store in -80C in a slide box.
I wrap the slide box in aluminum foil. Others use parafilm. Some put the slide box in a plastic bag and add dryerite.
For long term stability of antigens and mRNA (for ISH) what is the optimum way to take care of the slides? What actually is the goal? Should one try to reduce moisture (from freezer), hence the dryerite? Or is drying out the problem?
And when taking the slides out repeatedly, how should one handle them? Let the box come to room temp first so as to not allow moisture to condensate? Or should one move as quickly as possible?
I keep a collection slide box on dry ice. I cut slides and place slides in that box (on dry ice). Once slide box is full or ready for storage, I simply close the slide box and place at -80C. I do not think that wrapping the box with parafilm or plastic bag will give any additional protection.
Hi, Why do you let them (sections of fixed frozen brain and spinal cord tissues) dry for 10-15 mins at RT before storing in -80C?
@ Chandra - They adhere to the slide in that way. That is what I was taught at least.
Last time, a week ago, I took out slides from -80C and kept flattened at 4C over night and than 10 minutes at Room temp, washed with PBS in a jar, than blocking buffer etc, IF came excellent and sciatic nerve of mouse (1mm in dia or so) were well adhered to slide, small tip
Dear Alexander,
I think your method of fixation is appropriate. Defrosting the sections on Superfrost Plus slides at RT for 15-20 min can give a good attachment of the sections on slide surface. Regarding the storage of the sections, I would prefer to arrange the slides in a storage box and keep the storage box in a plastic bag. If you are storing your sections for longer duration, It would be a good idea to leave two open tubes of frozen MilliQ water in the bag to avoid the water loss from sections.
I think excess drying of the sections are not preferred for histology. So I would recommend to avoid the use of dryerite.
Cheers
Nimal Raveendran
I am doing similar assays for 10um brain sections, how long can you store these mounted slides at -80C? I see a lot of people say you can keep for ~3months but does it apply the same for 4%PFA-fixed mRNA as for proteins? Do we expect RNA to start todegrade after 3months?
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