[Histonet] Santa Cruz Antibodies
Hello Histonet, I'm hoping someone will be able to clear something up for me. I've seen several articles in the news recently regarding the revocation of Santa Cruz' animal dealer license, the closing of their research lab, and no longer being allowed to sell antibodies past December 31 of this year. I thought I had a good handle of what to expect, but my colleague just received an email the other day from Santa Cruz that they are expanding a bunch of products (monoclonal antibodies, RNAi, biochemicals and CRISPR). Did I miss something? Also, if you have any information, it would be great if you could include an official source. Thanks, Anna ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Samples not staining with Hematoxylin or primary
Thanks so much to everyone who responded! A couple people have suggested that this issue may be due to a disruption to the charge on the slide (due to high temp baking, water bath additives, etc) which causes a hydrophobic barrier to form and prevents reagent from reaching the tissue. This sounds very convincing to me, but we have found that repeated antigen retrieval using a pressure cooker (our standard retrieval for this particular primary) has allowed 2 of the 3 slides to stain with both DAB and Hematoxylin. Would repeated retrieval be able to reverse the slide charge back to normal? I'm having an issue with some recent samples and was wondering if anyone else has experienced the same thing...I stained some samples with a previously optimized protocol for anti-human Granzyme B on the Ventana Benchmark and about 1/3 of my samples did not stain at all with either GranzB or Hematoxylin. My controls and samples that did stain positive for GranzB look fine and the machine has been recently serviced and working well with other runs. I'd really appreciate your input if you have any advice or suggestions on what could have gone wrong. Thanks, Anna ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] IHC antibody/assay recommendations for IL-4, IL-13, IL-4R, and IL-13Ra1
Just wondering if anyone has had good luck with any of these antibodies (IL-4, IL-13, IL-4R, and IL-13Ra1) and would be willing to share vendor info and/or assay conditions they've tested? ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Samples not staining with Hematoxylin or primary
Hello Histonetters, I'm having an issue with some recent samples and was wondering if anyone else has experienced the same thing...I stained some samples with a previously optimized protocol for anti-human Granzyme B on the Ventana Benchmark and about 1/3 of my samples did not stain at all with either GranzB or Hematoxylin. My controls and samples that did stain positive for GranzB look fine and the machine has been recently serviced and working well with other runs. I'd really appreciate your input if you have any advice or suggestions on what could have gone wrong. Thanks, Anna ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Suggestions for staining bacteria in FFPE decaled growth plate
Happy Friday, Histonetters! I have a puzzle that I could really use your expert knowledge on this morning. One of our pathologists has a decalcified FFPE mouse bone embedded longitudinally and has found some bacteria located in the growth plate. The bacteria has been confirmed with fluorescence, but she really wants to be able to show the morphology of the tissue with the bacteria. So far, H, gram, Brown & Hopps stain the growth plate so dark that you can't make out the bacteria. My suggestions were to try a thinner section and adjust the timing of the H (haven't had a chance to do that yet), but I wanted to throw the question out to you all to see if anyone has tried other stains or techniques for this type of issue. I'd really appreciate the help! Thanks, Anna ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] HTL Career Path
Hello Histonet, I have been following this listserve for a couple years now and have found it to be a very interesting and helpful forum for sharing ideas and information among individuals with varied career histories in the histopath field. I sort of fell into histology work while working on my MS thesis in marine biology, making my own HE slides for diagnosis of a parasite found in blue crabs. Since I graduated in 2011, I've worked mostly in research histology labs and just got my HTL certification last spring, working on my qIHC this fall. I like the histology field and have had the pleasure of working with some really great people. However, I've felt discouraged at times with what seems like a lack of opportunity for professional growth and advancement and sometimes it feels like the only way to get new experiences and higher pay is to go back to school for another degree or completely change jobs and move to a new institution. I realize that I got into histology almost accidentally and that some of my experiences may be unique to the research field, but I'd be very interested to hear about your experiences regarding research vs. clinical work, how you were able to be intentional about your career path, what advancement opportunities you've found, etc. if you're willing to share them. Thanks, Anna ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Air bubbles in Coverslipper
Hi Histonetters! We have a Leica CV5030 coverslipper in our lab and we regularly (~ once a week or so) have trouble with large air bubbles showing up in a significant number of our slides. We have flushed the hose from the mounting medium through the needle with xylenes which sometimes works to clear the bubbles and sometimes does not. Has anyone else had trouble with this and possibly found a solution? I would really appreciate your advice! Thanks, Anna ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Histonet mailing list submission: Jones' Methenamine Silver Stain
Dear Histonetters, I was wondering if anyone has experience with Jones' Methenamine Silver stain? We have attempted this stain several times, each time with a slight tweak to the protocols we found online. We found that it took much longer than expected (3-4 hours compared to the expected 1 hour) for the membranes to develop the expected black color every attempt we made. The few times we did see the black color develop in the membranes, the next step in the gold chloride solution (1 minute) washed away all the dark color in the kidney, leaving it a very light brown. However, we did notice that only the skin sample on our test slides retained the black color. Since we ultimately need this stain for kidney samples, this didn't do us any good. We tried thinner sections (2 micron), acid washed glassware, brand new reagents, and multiple protocols with no luck. I would greatly appreciate any advice as to what we could try differently or if anyone else has experienced these same problems. Thank you! Anna ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
