[Histonet] uneven alternating sections on cryostat

Nathan Cramer natecrmr <@t> gmail.com
Mon Aug 31 16:02:20 CDT 2009


Thank you very much to everyone who has provided suggestions to help me 
fix my uneven section problem. I am preparing some more samples (spinal 
cord) at the moment and will see how things go later this week. I've 
summarized the suggestions below (in case someone else has the problem 
and finds this thread).

1. Check tightness of blade holder and be sure it is even in tightness 
on both sides.
2. Check tightness of screws/bolts pretty much everywhere including 
those that hold the cryostat to the cabinet and the chuck holder in place.
3. Check/adjust the blade angle. If it is too vertical, it can start 
cutting the front but compress the back of the specimen causing uneven 
sections.
4. There may be an issue with the cryostat head retracting too slowly. 
This could be due to my cutting too quickly or the need for a 
defrost/cleaning (by someone who knows what they are doing).
5. The orientation of the sample can influence how well the slices turn 
out. Changing the orientation may improve things.
6. Don't let the sample protrude too far away from the chuck.

If I find the smoking gun, I'll post an update...

Best wishes
Nate



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