[Histonet] iliac artery attachment to slides?

[Jack Ratliff]

Jim,
 
I think that I might better understand you now from your reply and would like to share my response with others that might be following this thread. From the looks of it you are using 50% EtOH on the slide before rolling out the section. You then mentioned that you once used a drop of xylenes instead of the 50% EtOH and everything works, but you lose sections during deplastification.
 
My routine procedure is to first lubricate the block during cutting with 70% EtOH, I then use 95% EtOH on the slide and periodically (if needed) on section during the process of rolling out the section and orientating on the slide, with the section wet with 95% EtOH I place a section cover over the top, then using bibulous paper and a brayer roller I gently start in the middle rocking back and forth while gradually pressing down with more force until I have attached the section to the slide. This typically help to eliminate wrinkles, however, with your type of specimens you can still have trouble so you need to improvise with the actually pressing method where you actually attach the section to the slide.
 
If you have not tried the higher concentrations of ethanol during the roll out steps, try it and let me know your results. Please keep in mind that I am assuming that you are using something like Haupt's Adhesive (Dorn and Hart Microedge) to coat your slides and that your resin blocks are fairly soft. You see, the higher alcohol concentration acts to help soften the resin so that it is easy to unroll the section and eliminate wrinkles. In fact, the concentration of ethanol and its effectiveness during this step is proportional to the hardness or softness of the resin block. I know that you use MMA + DBP + P-16, but can you tell us the concentrations? I use Acrylosin Infiltration & Embedding solution for my resin work now and the SOFT product contains 10% of the softening component.
 
One other thing to try with the attachment of these difficult type specimens is to float them out on the slide in a pool of 95% EtOH on a slide warmer. The 95% will help you to tease out the wrinkles and the heat of the slide warmer will help to evaporate the ethanol so that you can get some sort of initial attachment to the Haupt's before you try to press the section flat. Also, before you place the section cover on top of the section, add a drop of further diluted Haupt's to the section so that it can dry with the section just in case you are worried about gelatin coating loss. You might get some additional background, but at least it is something that you can tweak if the ethanol steps help to increase your ability to eliminate wrinkles.
 
Jack
 


 

> Date: Wed, 31 Aug 2011 09:04:28 -0500
> From: Herrick.James <@t> mayo.edu
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: FW: [Histonet] iliac artery attachment to slides?
> 
> 
> 
> Good morning everybody!!
> 
> I am trying to attach iliac arteries embedded in MMA (methyl
> methacrylate + dibutyl phthalate + perkadox 16) to gelatin coated slides
> to allow us to stain them with the Movat's Pentachrome. Unfortunately, I
> have a real problem getting them to attach - shortly after they are
> placed into 2-MEA for plastic removal, the section detaches from the
> slide.
> 
> I have mounted hundreds of sections to slides before with great success,
> but have never tried iliac arteries before. When mounting them using the
> conventional method (a drop of 50% ETOH, roll them onto the slide using
> a plastic coverslip, clamping the stack and placing it into a 45 - 50
> degree Celcius oven for 24 to 48 hours), the sections attach beautifully
> and do not fall off during deplasticization. The problem with the iliac
> sections is that they are completely consumed with wrinkles around the
> entire circular area of the artery, rendering them unuseable.
> 
> I have tried dry mounting (without using 50% ETOH or dH2O), I have tried
> floating the section onto a slide from a warm ETOH or dH2O solution (in
> water bath), I have tried using a slide warmer, I have tried a heat gun
> at low and high temperature settings, etc., etc.. The sections look
> great using the dry mounting methods (i.e. they don't wrinkle), but they
> detach from the slide very quickly following submersion in 2-MEA. It
> seems to me that the wrinkles appear, following the introduction of the
> ETOH or dH2O.
> 
> If anyone would have any suggestions or comments, it would be greatly
> appreciated. Thanks again for all of your help.
> 
> Have a great day!!
> Jim
> 
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