I've been trying to make 5 uM sections of injured mouse spinal cords and stain the tissue with Luxol Fast Blue and H&E. At first I was removing the bones from around the spinal cord, but the tissue just fell apart after I got it on the slide. I think because it had been severely damaged.
I thought I might be able to hold the tissue together better if I kept the bone, so I took a spine and decalcified it with 500mM EDTA for 1 week and tried again. But the bone tissue just pulled away from the spinal cord.
These were some of my better images. I have many others that are even less intact. I see the tissue fall apart on the slide during the drying step, after I put the section on the surface of a water bath and catch it on the slide. I have seen some sections spread all the way across the slide. I suspect it has something to do with water getting trapped under the paraffin section and not drying correctly.
How can I improve the procedure to keep the tissue in 1 piece?