[Histonet] new or used cryostat question?

Hello All, I'm starting my own lab and am trying to make the best use of my startup package. I section rodent brain on a sliding microtome with a dry ice stage (AO 860) but would like to move to fresh frozen sections mostly for RNA work and perhaps laser capture. I'm trying to decide whether it

[Histonet] AO-860 Sliding Microtome + OCT Embedded

I've tried something very similar with fresh brain tissue. I snap froze the tissue and then mounted it on the dry ice stage with little problems. But when I cut the tissue it thawed instantly on the knife and turned into a mess, even the thicker sections. You need to maintain the sections at a c

[Histonet] AO-860 Sliding Microtome + OCT Embedded Tissue?

Hi Francis, I don't know how thick your sections will be, but in general the AO-860 (fantastic microtome by the way) is not designed to cut fresh tissue. Certainly the relatively thick sections brain sections (50-200 micron) won't stand up very well in my experience. How thick are you trying to

[Histonet] used microscope vendors

Can anyone recommend a good source for either used microscopes or an inexpensive brand? I'm looking for a basic inverted microscope for very basic cell counts, and quick looks for cell viability and health. This is very much a quick in and out scope for students to kick around and abuse. I'm pur

[Histonet] how to fix floating sections?

using RNAlater-ice for example. Does anyone know if this screws up native fluorescence of GFP? Thanks! Caroline Bass ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] Cheap slide scanner for rat brains?

Hello Everyone, Does anyone have a good way to get low magnification images from slides? I have some DAB stained rat coronal sections that I would like to digitize. Basically, I want to have a picture of the entire section. It doesn¹t have to be high resolution, just enough to make out the basic s

[Histonet] Coverslipping video

Hello, I¹m wondering if anyone could direct me to an online video of coverslipping, preferably permount. I¹d like a reference to show some newbies in the lab. Thanks, Caroline ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.uts

[Histonet] Re: Histonet Digest, Vol 82, Issue 14

Any suggestions for staining nets for floating brain sections? I've looked at netwells, cell strainers and have even considered making my own. Most commercial options are way too expensive. I'd love to hear what everyone uses and if there is an easy way to make them. Thanks! On Sep 12, 2010,

[Histonet] Rnase free slides?

Hello, I'm doing RNA work for the first time. My plan is to take a fresh rat brain, block quickly, freeze by immersing in dry-ice cooled isopentane, storing at -80, collecting tissue sections (thickness will be determined, somewhere between 20 and 300 um), and punching the particular regions I nee

[Histonet] Used knife sharpeners

). Any recommendations on what to buy or look for in a used knife sharpener? Any recommendations for a book or website on basic knife sharpening techniques? Thanks, Caroline Bass ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http

[Histonet] AO 860 repairman in NC?

, Caroline Bass ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] 200 proof alcohol?

Hello, I¹m trying to buy some alcohol for both histology and molecular biology protocols. But I¹m having some difficult getting ³molecular biology grade² 100% ethanol. The hospital I work at sells 200 proof ethanol, but can¹t say anything about the quality. Since it¹s 200 proof, is that basically

[Histonet] EGFP IHC in floating brain sections

Hello, Sorry if this is a double post but I didn't see my first one in the digest. I¹m hoping someone could help me troubleshoot my staining protocol. I am staining to EGFP in microglia (produced by a virus I injected in the brain). Here¹s my general protocol: Deactive endogenous peroxidases (0.

[Histonet] EGFP IHC in floating brain sections

Hello, I¹m hoping someone could help me troubleshoot my staining protocol. I am staining to EGFP in microglia (produced by a virus I injected in the brain). Here¹s my general protocol: Deactive endogenous peroxidases (0.1M PB + 20% methanol + titon-x100 (80 ul in 40 ml total) + 0.3% h2O2) for 15