Re: [Histonet] removing MMA
We use acetone to dissolve MMA from implants. Don't know what that would do to tissue. Bob On Fri, May 6, 2011 at 11:15 AM, Bernice Frederick b-freder...@northwestern.edu wrote: We have a piece of bone in MMA. The researcher wants us to get It out and do paraffin. Is it possible? What will melt the MMA? We also have some 50um slides cut ,that they want to see blood vessels and collagen. Am I going to be able to do an EVG and trichrome or will the plastic inhibit this process? Thanks, Bernice Bernice Frederick HTL (ASCP) Senior Research Tech Pathology Core Facility ECOGPCO-RL Robert. H. Lurie Cancer Center Northwestern University 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 mailto:b-freder...@northwestern.edu b-freder...@northwestern.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Formaldehyde monitor
Anyone know of a cheap formaldehyde gas monitor or concentration test method? I found a couple of monitors on Google, but they seem to run abour $1500+ Bob Sepulveda VA Med Center Los Angeles ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] manual morphometric analysis
If you have a microscope camera of some kind, ImageJ is a free dowmloadable software package that will do morphometric analysis. See: http://rsbweb.nih.gov/ij/. There is a mailing list with a lot of helpful people on it too. Bob Nienhuis UCLA / VA Medical Center Los Angeles On Fri, Jun 25, 2010 at 2:59 PM, mohamed abd el razik k8...@yahoo.comwrote: hi all histonetters i need to do some morphomtric analysis on some samples of small intestin but we have no image analysis software so i need to do it manually by occular micrometer. i need to know how to calculate villus surface area? overall mucosal surface area of cross section? surface area of lamina propria? mitotic figure index?? i don't know how to caculate all . any help please thanks in advance mohamed faculty of vet. med. cairo univ.- egypt ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Spencer AO 820 microtome questions
How old are those 820s and 860s anyway? I know they were around in the early 70's. We still use our 860! Bob Nienhuis VA / UCLA Medical Center North Hills, CA On 3/16/10, Scott Parker spar...@vt.edu wrote: Dear all, I have access to a Spencer AO 820 microtome (a black beauty in Histo-parlance) that is does not have a disposable blade holder nor a chuck for holding paraffin cassettes. The microtome otherwise appears to be in good condition. I would like to get suggestions for where I might be able to purchase a chuck and disposable blade holder for this classic model. Alternatively, would it be a better idea for me to put this piece of equipment on display in a glass case and instead purchase a newer microtome. My work involves relatively low volume, basic sectioning of paraffin blocks for teaching and research. I don't need anything too fancy, just a microtome that is reliable and appropriate for student researchers to use. Thank you in advance for your responses. Scott Scott L. Parker, Ph.D. Assistant Professor Biology Coastal Carolina University P.O. Box 261954 Conway, SC 29528-6054 Office Phone: (843)-349-2491 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Spencer AO 820 microtome questions
Found an old message. The American Optical Co. that first made the 820 Microtome in 1947, is now part of Leica. Bob On Mon, Mar 22, 2010 at 9:50 AM, Paula Pierce cont...@excaliburpathology.com wrote: I am still using 5 820s!!! -- *From:* Bob Nienhuis bob.nienh...@gmail.com *To:* Scott Parker spar...@vt.edu *Cc:* histonet@lists.utsouthwestern.edu *Sent:* Mon, March 22, 2010 11:46:00 AM *Subject:* Re: [Histonet] Spencer AO 820 microtome questions How old are those 820s and 860s anyway? I know they were around in the early 70's. We still use our 860! Bob Nienhuis VA / UCLA Medical Center North Hills, CA On 3/16/10, Scott Parker spar...@vt.edu wrote: Dear all, I have access to a Spencer AO 820 microtome (a black beauty in Histo-parlance) that is does not have a disposable blade holder nor a chuck for holding paraffin cassettes. The microtome otherwise appears to be in good condition. I would like to get suggestions for where I might be able to purchase a chuck and disposable blade holder for this classic model. Alternatively, would it be a better idea for me to put this piece of equipment on display in a glass case and instead purchase a newer microtome. My work involves relatively low volume, basic sectioning of paraffin blocks for teaching and research. I don't need anything too fancy, just a microtome that is reliable and appropriate for student researchers to use. Thank you in advance for your responses. Scott Scott L. Parker, Ph.D. Assistant Professor Biology Coastal Carolina University P.O. Box 261954 Conway, SC 29528-6054 Office Phone: (843)-349-2491 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Fluorescent dyes and xylene
Will the Alexa Fluor dyes withstand some dehydration and clearing in alcohol and xylene? If they won't are there any bright fluorescent dyes that can be obtained congugated to a secondary antibody that will? Looking to immunostain tyrosine hydroxylase in rodent brain with a fluorescent marker in tissue that needs to be dehydrated and cleared, perhaps with an abbreviated series. Bob Nienhuis VA / UCLA Neurobiology Research ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Fluorescent dyes and xylene
Sorry, I should have mentioned that this will be done on thick frozen sections. Bob On Mon, Jul 20, 2009 at 10:37 AM, Bob Nienhuis bob.nienh...@gmail.comwrote: Will the Alexa Fluor dyes withstand some dehydration and clearing in alcohol and xylene? If they won't are there any bright fluorescent dyes that can be obtained conjugated to a secondary antibody that will? Looking to immunostain tyrosine hydroxylase in rodent brain with a fluorescent marker in tissue that needs to be dehydrated and cleared, perhaps with an abbreviated series. Bob Nienhuis VA / UCLA Neurobiology Research ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Autoflourescence
The Wright Cell Imaging Facility at University Health Network in Toronto has compiled information on reducing autofluorescence from the Histonet archives into a booklet. Very useful. http://www.uhnres.utoronto.ca/facilities/wcif/fdownload2.html Bob On Thu, Jan 15, 2009 at 1:36 PM, Patten, Nicole (NIH/NIAAA) [F] patte...@mail.nih.gov wrote: Hi- I am having a horrible time with autofluorescence in my human brain FFPE tissue. I have been using Sudan Black which helps a little, but the background is still pretty bad. The tissue is usually cut at 10-15um. Does anyone have any other suggestions? Has anyone tried photobleaching the tissue prior to staining with any success? I attempted this once with no luck... Any help would be very much appreciated! Thanks in advance. Nicole ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Histogel
Has anyone tried embedding a block of tissue like a mouse brain in Histogel, and then cutting frozen or on a vibratome? I am doing Golgi stain on mouse brain, and the tissue is VERY friable. Website says, No matter how small, friable, or viscous your histology or cytology specimen, HistoGel will encapsulate and retain the entire specimen during histological processing Bob Nienhuis UCLA / VA Medical Center On Tue, Jan 13, 2009 at 9:49 AM, Bartlett, Jeanine (CDC/CCID/NCZVED) j...@cdc.gov wrote: Histogel works great and does not absorb water like plain agar does. Jeanine Bartlett Infectious Diseases Pathology Branch (404) 639-3590 jeanine.bartl...@cdc.hhs.gov -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Monfils, Paul Sent: Tuesday, January 13, 2009 12:46 PM To: Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Processing 3-dimensional cell cultures into paraffin I have processed and embedded cells grown in/on agar, or post-embedded in agar or agarose, many times. It really isn't much different from processing and embedding tissue. You can trim the agar blocks to appropriate size with a scalpel (if necessary) either before or after fixation (if you trim them before fixation, do it while they are cold, so the agar is firm), and place them in a cassette just like a piece of tissue. Put them on the processor as usual, processing times the same as you would use for tissues of equivalent size, and embed as usual. One difference in sectioning however. Don't place the blocks in water after facing them off. The agar absorbs too much water and becomes soft. The blocks should be cut cold, but not moistened. Either place them on a cold plate without water, or put them in the refrigerator after facing, and take them out one at a time as you section them. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] RE: Histogel embedding mouse brain for cryotomy or vibratome
Currently doing Cryotomy on quickly frozen Golgi-Cox impregnated mouse brains at 100-120 microns. Cryoprotecting with 30% sucrose. Cutting at -15C. Tried fresh tissue and brief NBF perfusion(5 min). If I perfuse with NBF for 15 min, or immersion fix overnight, it seems to destroy spines and create artifact- possibly glial cells. Cutting much easier though! Might Histogel help? Bob Nienhuis UCLA / VA Medical Center On Tue, Jan 13, 2009 at 2:10 PM, gayle callis gayle.cal...@bresnan.netwrote: Bob, So which type of sectioning are you doing now? Cryotomy or vibratome? And how thick do you want the sections to be? If frozen sections on snap frozen mouse brain, simply turning the temperature up to -16C or so will make sectioning much easier and get rid of the shredding, friable sections. Also, are you fixing then sucrose cryoprotecting after NBF or 4% paraformaldehyde fixation of the brain (can be either perfused or immersion fixation)? Gayle Callis HTL(ASCP)HT,MT Bozeman MT ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
