[Histonet] quality sections of bone using tape transfer method
nto <@t> stowers.org
Tue Jul 30 10:37:14 CDT 2013
Recently, I have been spending much time using the tape transfer method to get sections of undecalcified mouse femur and humerus. After searching histonet archives, I am using some suggestions such as 4x coated slides, changing the knife angle, flashing several times, rolling the $#*! out of the sections (as was suggested :)) and letting the slides rest on dry ice after flashing them. Thank you because these have all helped the bone tissue to remain on the slide while pulling the tape off. Now my problem is the quality of the bone tissue. Microscopically, it looks like a cobble-stone walkway... many little circular structures all bumping up against each other. It is noticeable on unstained sections, so staining is not the cause. I have made sections between 3µ and 10µ and it is present on all, although the 3µ had one smooth area. I would compare it somewhat to a cryo section of brain that sections nicely, then crackles as it adheres to the slide. I have tried many different ways to get a better section, but have not yet hit on the magic mix. I am at the point of blaming the cryo jane apparatus since it is over 10 years old. We checked into changing the UV lamp but were told that parts are no longer made for this model. We are checking into a new piece of equipment, but first, is there anything I have overlooked? Has anyone seen this same type of feature I have described? Any thoughts as to what could be causing it and how to eliminate it?
Thank you in advance for any help and advice,
Stower's Institute for Medical Research
Kansas City, MO
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