Re: [Histonet] X-gal and pap pens?

From:Gayle Callis


Try the Vector ImmEdge pens, they are very clean, dry nicely, and have long 
since replaced Kiyota's pens in our lab.  Vector sells their pens in 
package of two, at a price of one pen from other vendors.   One thing to do 
when using the Immedge pen - shake the begolly's out of them to disperse 
the active marking goo.  Their technical rep recommended this be done.  I 
actually vortex mine to insure goo redistribution.

  It is interesting that Kiyota's Super Pap Pens were declared toxic.  I 
had wondered about this ( after noticing some very strong solvent odors) 
and asked them about it, but they said they were acceptable, but that fact 
changed somewhere along the line.

DAKO has a clear pen, but be careful when dispensing - if you press toooooo 
hard, you have a Niagara Falls of goo all over everything.  I macerated ( 
with a pliers!) their hard tip to make it a tidge fuzzier to capture the 
goo rush and also pulled a tip from a favorite pen and did a trade of hard 
tip with softer tip.   We also let DAKO pen marks dry longer, but I have 
that luxury of time and with frozen sections before staining and/or 
fixation.  Hopefully DAKO corrected this over-dispensing problem as the goo 
is good and it is clear.

If all else fails, you can buy hydrophobic barrier slides with lines in a 
circle, squares or rectangles that work quite well. Just mount the section 
inside lines and do the staining.  Erie has a huge selection of these, and 
we have even had custom hydrophobic barrier slides made for us to eliminate 
pap pens.  Lovely thing is the barrier stays put unlike the pen marking, 
and the lines we liked were the one that are flatter like the Erie control 
slides so mounting a coverslip is more flush with slide surface. You can 
always talk to Erie about their selection.

At 11:55 AM 12/2/2004, you wrote:
>I've searched far and wide for a solution to this problem, and I hope the 
>experts can finally help me. Our lab performs x-gal staining on tissue 
>sections. Prior to 6 months ago, we used Kiyota's Super Pap Pen (which put 
>down an excellent, CLEAR barrier). Our last pen ran (mostly) dry, and we 
>tried to order new pens from Kiyota. Apparently, their clear pens were 
>discontinued because of toxicity, and now the only pen available from them 
>is the Super HT Pap Pen. The new pens put down a flimsy, blue-green 
>colored barrier. The trouble is this - when X-gal staining, some component 
>of the stain reacts with the dye in the pap pen producing a grainy red 
>precipitate which discolors the slide, tissue, etc. In addition, an oily 
>reidue floats up on the solution and can stick to tissue while removing 
>it. In a nutshell, the new pens are awful. Does anyone know of a clear pen 
>that is still sold (EMS, zymed all seem to have colored pens), or have 
>experience with a colored pap pen that doesn'!
>  t react with xgal staining solution? Thanks
>Darren Hess
>MD/PhD student, Dept. of Neuroscience
>University of Pennsylvania School of Medicine
>Histonet mailing list

Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)

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