[Histonet] best fixative for eyes
What are people's thoughts/experience with different fixatives for eyes (rat, mouse and rabbit mostly). We currently use modified Davidson's solution, but the pathologist is considering going to Davidson's. I guess there has been some difficulty with cutting due to the lens not getting fixed well and causing problems. Thank you for sharing ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] re-using modified Davidson's solution
Hi All, Can one re-use modified Davidson's solution for fixing eyes and testes, or has it lost its potency/efficacy after being used previously? Can you rejuvenate it by adding more acetic acid? (how much?) I appreciate any helpful comments. Thank you, Robin Robin R. Dean, Ph.D. Senior Scientist Study Director Comparative Biosciences, Inc. 786 Lucerne Dr. Sunnyvale, CA (408) 738-8060 robin_d...@compbio.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] sections falling off slides
Hi All, We are trying to do IHC for multiple antigens on goat knees. The sections are decalcified FFPE sections and we are using a Dako autostainer. We are currently trying air-drying for 1-2 days and baking of tissue slides before use in IHC.. The one previous time we tried baking, we noticed a decrease in IHC signal. The sections are pretty large, we tried cutting them down, but can't decrease the size much. We also decreased the number of washes and size of washes on the Daok, which helped only slightly. Does anyone have any suggestions? Thank you in advance. Robin Robin R. Dean, Ph.D. Senior Scientist Study Director Comparative Biosciences, Inc. 786 Lucerne Dr. Sunnyvale, CA (408) 738-8060 robin_d...@compbio.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] silver stain on slide-mounted FFPE brain tissues
Hi Histonetters, We want to do a degeneration-selective silver stain on slide-mounted formalin-fixed paraffin-embedded monkey brain sections. It looks like most of the kits/protocols are for free floating tissue sections or chunks of tissue that are stained and then sectioned. Has anyone used this stain or a similar stain on slide -mounted brain sections? (something more like an IHC protocol). We need to do multiple different stains on sections from the same brain, so we don't want to silver stain a whole chunk of brain tissue, just some individual sections. I'd appreciate any suggestions or help anyone might have to offer. Thank you, Robin Robin R. Dean, Ph.D. Senior Scientist Study Director Comparative Biosciences, Inc. 786 Lucerne Dr. Sunnyvale, CA (408) 738-8060 robin_d...@compbio.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] use of destained sections in IHC?
Hi all, Is it possible to de-stain HE stained paraffin sections and re-use them for IHC staining??? Does anything special have to be done? Will it only work for some epitopes/stains? Does anyone have suggestions on how to do this? I am being asked to do this and don't want to waste my time if it won't work. I would appreciate any suggestions anyone might have Thank you, Robin Robin R. Dean, Ph.D. Senior Scientist Study Director Comparative Biosciences, Inc. 786 Lucerne Dr. Sunnyvale, CA (408) 738-8060 robin_d...@compbio.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] stains for visualizing new bone growth
Does anyone know of a good stain to use to clearly show new bone growth other than von Kossa stain? Would appreciate any suggestions anyone might have. Thank you, Robin Robin R. Dean, Ph.D. Senior Scientist Study Director Comparative Biosciences, Inc. 786 Lucerne Dr. Sunnyvale, CA (408) 738-8060 robin_d...@compbio.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Reg: Beta-2 minroglobin IHC
Hi all, We are trying to use beta-2 microglobulin IHC staining to ID human cells in goat joints that were engrafted with human cells some time ago and having nothing but trouble. Tissue was decalcified in immunodecal, formalin-fixed paraffin-embedded tissue. Beta-2 MG antibody is a polyclonal rabbit X human beta-2 antibody from Acris, and we are using DAKO Envision-HRP -con. secondary with DAB as chromogen. IgG isotype controls and DAB alone controls look clean. . Goat joints are falling off the charged slides with heat-induced citrate buffer antigen retrieval so went to enzymatic retrieval to keep them on the slide. o Does enzymatic antigen retrieval damage this epitope?? (previous successful uses of this antibody were with citrate antigen retrieval) o Any suggestions on how to keep sections on slides? Sections are kind of large, but pathologist wants that size. . Didn't get hardly any staining at 2.5 ug/ml of the beta-2MG antibody, but at 5 ug/ml everything stains including all of goat joints. . Are there other or better stains that are used for identifying human cells engrafted into other species (label uniformly most human cells)? I saw mention of some mitochondrial antgen antibodies but no specifics. . Does Beta-2 antibody label only human cells? Seems like there are homologous markers in other species that may label. Currently we aren't using any blocks except peroxidase block. Thought we might try a CAS block to see if that will prevent sticking? To non-human tissue. Any suggestions or help with these problems will be greatly appreciated. We have to get the stain to work and are running out of ideas. Robin Robin R. Dean, Ph.D. Senior Scientist Study Director Comparative Biosciences, Inc. 786 Lucerne Dr. Sunnyvale, CA (408) 738-8060 robin_d...@compbio.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
