RE: [Histonet] zebrafish embryos histology
Patty, I have personally never performed histology on zebra fish embryo's, but if I was to test it out on my own I might try using the JB4 Plus GMA kit. It just seems to me that paraffin might cause too much shrinkage and maybe using an MMA protocol might be too harsh on the tissue. Besides, given the hydrophilic nature of GMA it just might be the best overall solution. Best, Jack From: pl...@uab.edu To: histonet@lists.utsouthwestern.edu Date: Wed, 22 Jan 2014 15:00:40 + Subject: [Histonet] zebrafish embryos histology Can anyone give me a method for zebra-fish embryo histology? The papers I've read show photos, but no description of histology in M M. I need to put several embryos in each block, and get the orientation correct, and put multiple sections on each slide, in hopes of getting one or two that are perfect. Any suggestions would be greatly appreciated. Thanks, Patty Lott UAB CMBD Core Lab 205-934-2007 ___ 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] zebrafish embryos histology
Patty, did some zebrafish work years ago, either pure histology or sections after we did whole mount ISH on the embryo's. As per Jack Ratliff's post, paraffin I found was really tough for orientation and for getting enough sections of such small embryo's. But would use, as Jack suggested, JB-4 Plus and would get beautiful sections, many embryo's in a block and multiple sections per slide. What I would do is under a dissecting scope would, with a VERY fine tool, push the multiple embryo's into an ordered row with similar orientation in the unpolymerized GMA block. Polymerize. If I wanted longitudinal sections, cut the block as is. If we wanted cross-sections, just gross cut the polymerized block and put it in a second GMA block of unpolymerized GMA and stand it up so the embryo's were on end and polymerize . Was every individual embryo correct? No! But enough were so got great HE sections or also seeing the ISH probe revealed at the cellular level. Ray, retired in Seattle - Original Message - From: Patricia F Lott pl...@uab.edu To: histonet@lists.utsouthwestern.edu Sent: Wednesday, January 22, 2014 7:00:40 AM Subject: [Histonet] zebrafish embryos histology Can anyone give me a method for zebra-fish embryo histology? The papers I've read show photos, but no description of histology in M M. I need to put several embryos in each block, and get the orientation correct, and put multiple sections on each slide, in hopes of getting one or two that are perfect. Any suggestions would be greatly appreciated. Thanks, Patty Lott UAB CMBD Core Lab 205-934-2007 ___ 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] zebrafish embryos histology
Hi Patty, You sort of piqued my curiosity of what is going on in the zebra fish world and found this youtube, 6 minute film http://www.youtube.com/watch?v=kZDwo20hl1Efeature=youtu.be About what is going on at Welcome Trust Research on zebra fish but maybe you know all this already. Anyway, I think the film is pretty neat with some incredible (not histology) but CONFOCAL slices through the fish. We had success doing this by confocal on whole-mount but again when studying gene manipulation and cell signal distribution and wanting histology in embryo's, had better luck with GMA. Paraffin as mentioned can certainly be done and is certainly easier but we could never get the resolution or number of sections we desired using paraffin. Maybe the people in lab from this film can give you some suggestions. Ray, retired in Seattle - Original Message - From: koelli...@comcast.net To: Patricia F Lott pl...@uab.edu Cc: histonet@lists.utsouthwestern.edu Sent: Wednesday, January 22, 2014 8:02:24 AM Subject: Re: [Histonet] zebrafish embryos histology Patty, did some zebrafish work years ago, either pure histology or sections after we did whole mount ISH on the embryo's. As per Jack Ratliff's post, paraffin I found was really tough for orientation and for getting enough sections of such small embryo's. But would use, as Jack suggested, JB-4 Plus and would get beautiful sections, many embryo's in a block and multiple sections per slide. What I would do is under a dissecting scope would, with a VERY fine tool, push the multiple embryo's into an ordered row with similar orientation in the unpolymerized GMA block. Polymerize. If I wanted longitudinal sections, cut the block as is. If we wanted cross-sections, just gross cut the polymerized block and put it in a second GMA block of unpolymerized GMA and stand it up so the embryo's were on end and polymerize . Was every individual embryo correct? No! But enough were so got great HE sections or also seeing the ISH probe revealed at the cellular level. Ray, retired in Seattle - Original Message - From: Patricia F Lott pl...@uab.edu To: histonet@lists.utsouthwestern.edu Sent: Wednesday, January 22, 2014 7:00:40 AM Subject: [Histonet] zebrafish embryos histology Can anyone give me a method for zebra-fish embryo histology? The papers I've read show photos, but no description of histology in M M. I need to put several embryos in each block, and get the orientation correct, and put multiple sections on each slide, in hopes of getting one or two that are perfect. Any suggestions would be greatly appreciated. Thanks, Patty Lott UAB CMBD Core Lab 205-934-2007 ___ 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
