And other crazy stuff. RE: [Histonet] cutting honey bees

Tue Jan 6 16:38:30 CST 2015

The coolest thing I cut was 600 yo deer bone.  One of our pathologist did archeology as a hobby and wanted me to section it, it was petrified as hard as a rock.  We tried everything to soften it to no avail.  We ended up cutting with a diamond wire lapidary saw without embedding it in anything.  Could make sections as thin as 30 microns.  Had a heck of a time trying to get sections to adhere to a glass slide and the sections would not take any stain.  We did end up looking at it with a fluorescent scope.  We could see rings like a tree of bone turn over.  This is how tetracycline labeling of bone first came about.  Someone way back stuck a piece of dinosaur bone under UV light and saw rings, apparently the animals eat moldy grain ( tet is made from mold) and it deposits where ever new bone is being laid down, it also happens to fluoresce.  I spent 25 years in a metabolic bone disease lab, we treated the patient with a course of tetracycline then waited for a period of time, I think it was 10-14 days, then the patient took another dose of tet and then within a day or 3 we biopsied their bone usually from the illiac  crest fixed it in methanol because the tetracycline was water soluble then processed it into GMA plastic without decal.  Unstained 5 micron sections cut with a tungsten carbide blade were reviewed with a fluorescent scope revealing the two labels of tet, since we knew the time between doses we could measure the area between the two labels and report them out as bone growth in mm per day.  People with severe lack of bone turn over would just have one single label meaning they were not making much bone.  

Cheers,
Patsy

Patsy Ruegg, HT(ASCP)QIHC
Ruegg IHC Consulting
40864 E Arkansas Ave
Bennett, CO 80102
H 303-644-4538
C 720-281-5406
pruegghm <@t> hotmail.com

> From: rjr6 <@t> psu.edu
> To: Timothy.Morken <@t> ucsf.edu; pruegghm <@t> hotmail.com; classicdoc <@t> gmail.com; histonet <@t> lists.utsouthwestern.edu
> Subject: RE: And other crazy stuff.  RE: [Histonet] cutting honey bees
> Date: Tue, 6 Jan 2015 20:54:10 +0000
> 
> The oddest things I cut were the honey bee and yellow jacket stingers.  I've done plant stamen, reptiles, fish and I believe another insect.  I usually tell the students that are working on a research project to give me a sample they don't care about so I can see if I can do what they want.
> 
> But I had oddities that I didn't have to section like during hunting season a hunter killed a deer and there was a mass on the trachea that he wanted tested to make sure the deer was okay to eat.  I got the sample and when I tried to gross it I found a very hard shiny silver object.  I told the pathologist whose case it was that the mass was from a bullet did he still want histo done. No.
> 
> The other interesting one was the egg shell.
> The conversation went something like this.
> Pathologist:  Can you section this egg shell
> Me: No it's too hard.
> P: Can't you decal it
> M: That's not going to work.
> P: Did you try.
> M: No
> P: Don't you think you should try first.
> M: Okay fine but it is no going to work.
> 
> Put a piece of eggshell (made of calcium) into some decal solution (that removes calcium) and watch the egg shell bubble and disappear.  I did get to tell the pathologist "I told you so"
> 
> Roberta Horner
> Penn State University
> Animal Diagnostic Lab
> 
> -----Original Message-----
> From: Morken, Timothy [mailto:Timothy.Morken <@t> ucsf.edu] 
> Sent: Tuesday, January 06, 2015 2:24 PM
> To: Patsy Ruegg; Roberta Horner; Douglas Gregg; Histonet <@t> Lists. Edu
> Subject: And other crazy stuff. RE: [Histonet] cutting honey bees
> 
> You crazy research people...OK, so what is the craziest thing you ever had to cut, or were asked to cut?
> 
> For me, not too bad, but embedding for EM and sectioning a single oocyte that was nearly microscopic. I'll just say it took a LOT of thick sections too face down to it without actually cutting through it.
> 
> 
> Open the floodgates....
> 
> Tim Morken
> 
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Patsy Ruegg
> Sent: Tuesday, January 06, 2015 11:13 AM
> To: Roberta Horner; Douglas Gregg; Histonet <@t> Lists. Edu
> Subject: RE: [Histonet] cutting honey bees
> 
> for the whole bee I probably would process and embed it in glycol methacrylate (gma) it is much harder and would give better sections, we have done zebra fish and several other harder tissues including calcified bone in GMA.
> 
> Cheers,
> Patsy
> 
> Patsy Ruegg, HT(ASCP)QIHC
> Ruegg IHC Consulting
> 40864 E Arkansas Ave
> Bennett, CO 80102
> H 303-644-4538
> C 720-281-5406
> pruegghm <@t> hotmail.com
> 
> 
> 
> > From: rjr6 <@t> psu.edu
> > To: classicdoc <@t> gmail.com; histonet <@t> lists.utsouthwestern.edu
> > Date: Sat, 3 Jan 2015 23:15:33 +0000
> > Subject: RE: [Histonet] cutting honey bees
> > CC: 
> > 
> > I sectioned and stained honey bee and yellow jacket stingers years ago.  They wanted to show the difference between the stingers.  I wasn't sure what to do so I processed and handled like everything else.  I was able to get some good sections.  I put 6 stingers in each block and cut several sections figuring there should be at least one good stinger in each block and it worked.
> > Roberta Horner
> > Penn State University
> > Animal Diagnostic Lab
> > ________________________________________
> > From: histonet-bounces <@t> lists.utsouthwestern.edu
> > [histonet-bounces <@t> lists.utsouthwestern.edu] on behalf of Douglas Gregg 
> > [classicdoc <@t> gmail.com]
> > Sent: Saturday, January 03, 2015 6:08 PM
> > To: histonet <@t> lists.utsouthwestern.edu
> > Subject: [Histonet] cutting honey bees
> > 
> > Has anyone had experience embedding and cutting honey bees. I am sure 
> > there are some issues with the harder exoskeleton. Would that have to 
> > be dissected away first. I am considering helping a student with a 
> > science fair project on bees.
> > 
> > Douglas Gregg
> > Veterianary pathologist
> > 
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> > Histonet <@t> lists.utsouthwestern.edu
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