[Histonet] bone paraffin embedding
Histonetters: Our lab needs to paraffin embed and cut bone. Is there a special process for fixation of bone, or can it be harvested and dropped right into NBF? -- Nicole Cosenza Research Technician Institute for Plastic Surgery SIU School of Medicine Springfield, Il 217.545.3862 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] lyophilizing company
Histonetters: My lab is needing to lyophilize some skin tissue. Is there a company that offers such services? ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] FISH technique and NBF as fixative
I need to do FISH with the universal bacterial probe EUB338. I am not too familiar with FISH. Can we fix the sample with NBF or is 4% paraformaldehyde better? The samples are acellular dermal matrix. A quick lit search shows people using 4% paraformaldehyde, but the histologist that will be embedding the tissue said that NBF is gentler on tissue than 4% PFA... ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] needing nerves resin embedded
Hi all: My lab is starting a project involving neuromas. We are in need of resin embedding of the nerves, but our facility does not have the capabilities. Is there a company that offers resin embedding/sectioning services? ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] cryosections with DiD labeling
We are beginning a retrograde tracing study in rats. We injected DiD into the rats and in 4 weeks we will perfuse and harvest the brainstems. My question is, is vibratome sectioning better than cryostat sectioning? The literature that accompanied the dye said either is an option, but some have noted dye leaching out when cyrostat sectioned. How common is this problem? If vibratome is the better choice, is there a good embedding protocol? ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] re-usability of ethanols
Hello all, I am looking for opinions on how long/often I can re-use ethanols (100%, 95%, 50%) when I rehydrate paraffin sections for IHC staining. Thanks! ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] need a counter stain for esterase and silver stained slides
I have recently stained some muscle tissue with bromoindigo and urea-silver to examine the neuromuscular junction. The nerve axons and endplates stain quite well this way, but I'd like to add some color to the slide for picture-purposes. Is there a good counterstain (eosin, or Biebrich scarlet-acid fuchsin) I can dip the slides in to add color, WITHOUT taking away from the stain already present in the slides? ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] combined cholinesterase-silver stain
I am looking into staining motor end plates. I've come across this combined cholinesterase-silver stain (reference Pestronk and Drachman, 1978). Based on the date of the paper, I'm wondering what the current technique is for this double staining. Anyone currently doing AchE and axon staining on fresh frozen muscle sections? ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Karnovsky and Roots stain
I am looking into a project involving motor end plate staining. Literature that I've found continually references Karnovsky and Roots from the 60s. However the papers are not supplying all the details. Does anyone do AchE staining by this method on fresh frozen, unfixed tissue sections? If so, can I get a more detailed protocol (fixation steps, washes, etc)? ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] DiD retrograde tracing study. Vibratome or cryostat
We are beginning a retrograde tracing study in rats. We injected DiD into the rats and in 4 weeks we will perfuse and harvest the brainstems. My question is, is vibratome sectioning better than cryostat sectioning? The literature that accompanied the dye said either is an option, but some have noted dye leaching out when cyrostat sectioned. How common is this problem? If vibratome is the better choice, is there a good embedding protocol? Nicole Southern Illinois University School of Medicine ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
