[Histonet] RE: MMA Mounting Issue with Haupts Adhesive

[Thomas, Nancy]

Jesse,
I have discovered that there are many different ways to work with MMA.  Probably no two techs are doing it exactly the same because there are so many steps in which to optimize it for your lab.  From reading your protocol, I see similarities and differences.  Rather than saran wrap, I use a thicker plastic that I cut into strips ( the size of the slide) from the plastic bags that we use for discarding paraffin waste.  Although this is different from your protocol, I still think that what I do to remove the plastic might work for you:  just before removing the plastic strip, I shoot it with a blast of freeze spray.  The plastic just about falls off.  

Nancy Thomas
Stowers Institute for Medical Research
Kansas City, MO

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Hernandez, Jesus Willibaldo
Sent: Wednesday, December 03, 2014 11:24 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] MMA Mounting Issue with Haupts Adhesive

Hi all,

I had a question about how to properly mount MMA sectioned slides using Haupt's (Dorn and Hart Microedge). My protocol is as follows:


1.       Stick Glass Slide into 50mL Falcon Tube with Haupts solution (1:1 50% EtOH:50% Haupts).

2.       Rotate the tube to ensure entire coating of glass slide.

3.       Let glass slide dry in fume hood for about 10minutes.

4.       Remove cut microtome sections (range: 50-5microns).

5.       Place cut sections in 70%EtOH to soften the plastic and unroll. (sections are coming out rolled up like scrolls)

6.       Remove a section from the 70%EtOH using forceps and try unrolling it with a needle on a glass slide.

7.       Once the section(s) is stretched on the coated glass slide, cover with saran wrap.

8.       Cover the wrapped slide with coarse filter paper and use a small seam roller to roll the sections on the glass slide as flat as possible. This will also help remove excess 70%EtOH.

9.       Remove the filter paper and stack glass slides in a slide press or similar device. Place tongue depressors or small pieces of wood equivalent to the slide length that are about 1cm thick at the end of the glass slide stack.

10.   Let slides dry over night at room temperature in press.

11.   Remove the slides from the press and unwrap the saran wrap to expose your adhered sample.

Questions/Comments:


1.       When I pull off the saran wrap, the sample sometimes comes off. The plastic in the sample turns white and dries out and does not attach. This is seen in the majority of my tissue sections.

2.       The only time I have noticed consistent attachment is for a 5micron sized section.

3.       How can I adjust the microtome so that the samples do not come off so scroll-like? My cutting angle is typically either 0 or 2.5 degrees. I also apply 70% EtOH to the block surface to soften the plastic.

Overall goal:
Get the tissue sections adhered to the glass slides so that I can proceed with staining. Thank you all.

Regards,

Jesse Hernandez


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