I am sectioning human / mouse tissue on a vibratome. At both 50um and 20um, the sections float off in surrounding PBS still with the surrounding agar layer attached, and I can't seem to easily remove it without disturbing the tissue.
This happens at both 4% and 10% agar. I have been using wet ice surrounding the PBS-filled vibratome chamber to keep everything cool, since at room temperature the sections did not cut well.
Has anyone else experienced this, or does anyone have any suggestions? Thank you.
I usually remove agar after transferring sections on the glass slide. I get rid of excess PBS with paper towel so my sections could hold on the glass. And then I make 1-3 cuts in agar layer only, with scalpel or blade (I mean radial cuts from tissue edge to periphery of the agar), so they could be easily removed with forceps.
Also, you can try 2% agar blocks. That's how I cut mouse brains.)
Chris Taylor
Hi there! I'm Abby, Chief Scientific Officer at Precisionary Instruments, where we've been crafting Compresstome vibratomes and microtomes for nearly two decades. With a wealth of experience in tissue sectioning, I'm here to help!
Regarding your query about the agarose rim, there are a couple of tricks I'd recommend. First, consider a sucrose dip for your tissue specimen into a 30% sucrose solution before embedding it into agarose. Alternatively, you can gently brush some sucrose onto the tissue. Additionally, try increasing the agarose concentration, going as high as 3%. These two techniques should aid in making that agarose rim "fall off" more smoothly. Hope this helps!
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