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Author Topic: Triton & hydrophobic pen  (Read 16294 times)

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Offline dangdangback

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Triton & hydrophobic pen
« on: July 27, 2005, 12:11:26 PM »
I use hydrophobic pen (Lipshaw) to circle my tissue. It works well without trion.

However, whenever I add triton, the hydrophobic circle is dissolved, and the tissue may get dry.

Any tricks to solve this problem?

Triton & hydrophobic pen
« on: July 27, 2005, 12:11:26 PM »

Offline richard03

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Triton & hydrophobic pen
« Reply #1 on: July 27, 2005, 08:47:47 PM »
I had similar problem before. I end up switching to MicroProbe staining system. It works excellently.  Alternatively you can use  Shandon Sequenza Manual Immunostainer and it works very well too.

Richard

Offline richard03

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Triton & hydrophobic pen
« Reply #2 on: July 27, 2005, 08:49:25 PM »
Just another thought. You can also use plastic coverslip for incubation.

Offline dangdangback

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Triton & hydrophobic pen
« Reply #3 on: July 27, 2005, 09:47:22 PM »
Quote from: "richard03"
Just another thought. You can also use plastic coverslip for incubation.


You mean put plastic coverslip on, and take it away between each steps? Will my tissue stay on the slide when I take the coverslip away?

Offline dangdangback

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Triton & hydrophobic pen
« Reply #4 on: July 27, 2005, 09:49:12 PM »
I never use "MicroProbe staining system", or "Shandon Sequenza Manual Immunostainer "

I wonder if I could persuade my PI to buy that. Are they expensive, and easy to do?

Offline richard03

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Triton & hydrophobic pen
« Reply #5 on: July 27, 2005, 10:27:57 PM »
Yes. You basically put plastic coverslip on for incubation and take it away during rinsing. It will not damage tissue. However I found it is very time consuming and inefficient.

The MicroProbe System will probably cost $300 per holder for 20 slides. You will need to purchase additional staining trays.

Shandon Sequenza will cost around 200 per tray and additioanl cost for plate. I found this is a better deal and works excellently for IHC.

Richard

Offline excalibur

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Triton & hydrophobic pen
« Reply #6 on: July 31, 2005, 03:41:19 PM »
I agree with Richard that the Shandon Sequenza is an excellant way to perform IHC by hand. You do not need to purchase the entire system. The racks and coverplates are sold separately. I even wash the coverplates in soapy bleach water and reuse them. You can find them at Thermo-Shandon's website.
Paula Keene Pierce, HTL(ASCP)HT
Excalibur Pathology, Inc.
5830 N Blue Lake Dr.
Norman, OK 73069
405-759-3959
www.excaliburpathology.com

Offline ImmunoNYC

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Triton & hydrophobic pen
« Reply #7 on: August 01, 2005, 12:45:12 AM »
Or just try a pap pen from a different source. I am picky about the pap pens I use as I find huge differences between different vendors. My personal favorite is the Mini Pap Pen made by Zymed. It is indestructable. Other alternatives are real wax pencils or buy slides with hydrophobic chambers.

Offline discman

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Triton & hydrophobic pen
« Reply #8 on: August 11, 2005, 11:40:00 AM »
What we often use is a piece of Parafilm instead of the plasctic coverslip. The parafilm is very easy to remove and works as fine as the plastic coverslip. It's also cheaper I guess.

Offline Karolina

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Triton & hydrophobic pen
« Reply #9 on: November 07, 2005, 07:56:29 AM »
The idea with parafilm is great :) To take parafilm gently from the slide (so as not to damage the tissue) I would reccomend to "inject" 100 ul of PBS (or buffer used) between tissue and parafilm - goes easier :)

Offline saar

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Re: Triton & hydrophobic pen
« Reply #10 on: February 23, 2011, 05:05:21 AM »
I'm using the GnomePen from invignome (see www.invignome.com).
It works better, it doesn't get eaten away by the Triton X100 or Tween20 and so on and it's color coded so you can avoid mistakes like adding the primary antibody to your negative control...
Hope it helps.
Saar
 ???

Offline MT Scientist

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Re: Triton & hydrophobic pen
« Reply #11 on: February 23, 2011, 05:20:31 PM »
Sounds like you need to do your incubations in a moist chamber (a tupperware container with moistened paper towels in the bottom and your slides on racks will work).

Re: Triton & hydrophobic pen
« Reply #11 on: February 23, 2011, 05:20:31 PM »