Re: [Histonet] bat wing histology
Hi, We did this on derm punch biopsies for a while. We used PLP (paraformaldehyde-lysine periodate) fixative. This was also done on a sliding microtome at 50um with piles of dry ice to keep the sucrose frozen. It was a bit of a pain in the butt, but we got decent results. I think the sectioning process could have been easier, but the PI was obsessed with not modifying the project at all, even if it meant improvements. We stained these as floating sections in 96 well plates, so in this case I would be concerned that the wing rolls would un-roll if you were to do the swiss roll thing I described. Have fun :-) Amos On Tue, Jul 6, 2010 at 5:10 PM, Mohit Chadha wrote: > Thank you everyone for replying, much appreciated. > > Having also talked to people in my dept, I have a rudimentary protocol > ready. Of course, I will have to tweak it to see what works. I will be using > anti PGP9.5 antibody for neuronal immunology. > > I am still not sure how to section the wing. In most likelihood, I will be > using freezing sliding microtome. The "swiss roll" method sound good and I > will definitely try it. I am thinking that since the wing membrane is thin > (~30 um), I will also try to use the whole mount of small pieces. > > Any other thoughts and advice would be appreciated. > > Thank you, > Mohit Chadha, > Univ of Maryland. > > > > > > > > On Fri, Jul 2, 2010 at 4:14 PM, Amos Brooks wrote: > >> Hi, >> I do hope you are looking at cross sections of the wing and not the >> flat. That would be very difficult indeed. For good cross sections I would >> try a "Swiss Roll". This is a way of demonstrating a large amount of cross >> sectional area in small space. Take the membrane and fix it by submersion in >> the fixative of your choice. Prior to processing roll the whole membrane up >> then cut the membrane log into sections small enough to fit in a cassette. >> You can use foam biopsy pads to support this shape. Embed it and section it >> on edge to show a long coiled membrane. The hairs should be able to be >> displayed in this way as well. To show a lot of membrane at the same time >> you could place multiple rolls in one cassette. This should work well. >> >> Good Luck, >> Amos >> >> >> Message: 21 >> Date: Thu, 1 Jul 2010 11:46:17 -0400 >> From: Mohit Chadha >> Subject: [Histonet] bat wing histology >> To: histonet@lists.utsouthwestern.edu >> Message-ID: >> >> Content-Type: text/plain; charset=ISO-8859-1 >> >> Hello everyone, >> >> This is my very first post and I am desperately looking for help. I am new >> to histology, so any help would be much appreciated. >> >> I am studying the peripheral sensory innervation of bat wings. As a first >> step, I would like to demonstrate the innervation pattern on the different >> parts of the wing membrane (a whole mount of the wing?). Second, I would >> like to demonstrate the mechanoreceptor make-up of the tiny hairs on the >> wing membrane. >> >> Bat wings are highly elastic, with numerous folds, a thickness of about >> 35-45 microns (in the species I study), a network of thin collagen >> bundles, >> and pigmented superficial epidermal layers. >> I could provide more information if required. >> >> Hoping to hear back from the members. >> Thank you. >> > > ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] bat wing histology
Thank you everyone for replying, much appreciated. Having also talked to people in my dept, I have a rudimentary protocol ready. Of course, I will have to tweak it to see what works. I will be using anti PGP9.5 antibody for neuronal immunology. I am still not sure how to section the wing. In most likelihood, I will be using freezing sliding microtome. The "swiss roll" method sound good and I will definitely try it. I am thinking that since the wing membrane is thin (~30 um), I will also try to use the whole mount of small pieces. Any other thoughts and advice would be appreciated. Thank you, Mohit Chadha, Univ of Maryland. On Fri, Jul 2, 2010 at 4:14 PM, Amos Brooks wrote: > Hi, > I do hope you are looking at cross sections of the wing and not the > flat. That would be very difficult indeed. For good cross sections I would > try a "Swiss Roll". This is a way of demonstrating a large amount of cross > sectional area in small space. Take the membrane and fix it by submersion in > the fixative of your choice. Prior to processing roll the whole membrane up > then cut the membrane log into sections small enough to fit in a cassette. > You can use foam biopsy pads to support this shape. Embed it and section it > on edge to show a long coiled membrane. The hairs should be able to be > displayed in this way as well. To show a lot of membrane at the same time > you could place multiple rolls in one cassette. This should work well. > > Good Luck, > Amos > > > Message: 21 > Date: Thu, 1 Jul 2010 11:46:17 -0400 > From: Mohit Chadha > Subject: [Histonet] bat wing histology > To: histonet@lists.utsouthwestern.edu > Message-ID: > > Content-Type: text/plain; charset=ISO-8859-1 > > Hello everyone, > > This is my very first post and I am desperately looking for help. I am new > to histology, so any help would be much appreciated. > > I am studying the peripheral sensory innervation of bat wings. As a first > step, I would like to demonstrate the innervation pattern on the different > parts of the wing membrane (a whole mount of the wing?). Second, I would > like to demonstrate the mechanoreceptor make-up of the tiny hairs on the > wing membrane. > > Bat wings are highly elastic, with numerous folds, a thickness of about > 35-45 microns (in the species I study), a network of thin collagen bundles, > and pigmented superficial epidermal layers. > I could provide more information if required. > > Hoping to hear back from the members. > Thank you. > ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] bat wing histology
Hi, I do hope you are looking at cross sections of the wing and not the flat. That would be very difficult indeed. For good cross sections I would try a "Swiss Roll". This is a way of demonstrating a large amount of cross sectional area in small space. Take the membrane and fix it by submersion in the fixative of your choice. Prior to processing roll the whole membrane up then cut the membrane log into sections small enough to fit in a cassette. You can use foam biopsy pads to support this shape. Embed it and section it on edge to show a long coiled membrane. The hairs should be able to be displayed in this way as well. To show a lot of membrane at the same time you could place multiple rolls in one cassette. This should work well. Good Luck, Amos Message: 21 Date: Thu, 1 Jul 2010 11:46:17 -0400 From: Mohit Chadha Subject: [Histonet] bat wing histology To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Hello everyone, This is my very first post and I am desperately looking for help. I am new to histology, so any help would be much appreciated. I am studying the peripheral sensory innervation of bat wings. As a first step, I would like to demonstrate the innervation pattern on the different parts of the wing membrane (a whole mount of the wing?). Second, I would like to demonstrate the mechanoreceptor make-up of the tiny hairs on the wing membrane. Bat wings are highly elastic, with numerous folds, a thickness of about 35-45 microns (in the species I study), a network of thin collagen bundles, and pigmented superficial epidermal layers. I could provide more information if required. Hoping to hear back from the members. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] bat wing histology
Make that an open discussion on the list, no "under cover posting" if possible, please! :) > Hello everyone, > > This is my very first post and I am desperately looking for help. I am new > to histology, so any help would be much appreciated. > > I am studying the peripheral sensory innervation of bat wings. As a first > step, I would like to demonstrate the innervation pattern on the different > parts of the wing membrane (a whole mount of the wing?). Second, I would > like to demonstrate the mechanoreceptor make-up of the tiny hairs on the > wing membrane. > > Bat wings are highly elastic, with numerous folds, a thickness of about > 35-45 microns (in the species I study), a network of thin collagen bundles, > and pigmented superficial epidermal layers. > I could provide more information if required. > > Hoping to hear back from the members. > Thank you. > ___ > 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] bat wing histology
Hello everyone, This is my very first post and I am desperately looking for help. I am new to histology, so any help would be much appreciated. I am studying the peripheral sensory innervation of bat wings. As a first step, I would like to demonstrate the innervation pattern on the different parts of the wing membrane (a whole mount of the wing?). Second, I would like to demonstrate the mechanoreceptor make-up of the tiny hairs on the wing membrane. Bat wings are highly elastic, with numerous folds, a thickness of about 35-45 microns (in the species I study), a network of thin collagen bundles, and pigmented superficial epidermal layers. I could provide more information if required. Hoping to hear back from the members. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
