[Histonet] zebrafish embryos histology

2014-01-22 Thread Patricia F Lott
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
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RE: [Histonet] zebrafish embryos histology

2014-01-22 Thread Jack Ratliff
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
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Re: [Histonet] zebrafish embryos histology

2014-01-22 Thread koellingr
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 
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Re: [Histonet] zebrafish embryos histology

2014-01-22 Thread koellingr
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 
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[Histonet] zebrafish and IHC

2013-02-20 Thread Margaryan, Naira
Dear Histonetters,

I am trying to do  IHC on FFPE 4 weeks old zebrafish with different Abs. Is 
there a trick working with zebrafish? I am using the same IHC protocol I always 
use on human and mouse tissue and my Abs are  suppose to work on fish as well 
as human. I run human and fish sections together with same AR and IHC protocol. 
By the end I get beautiful staining on human section and or nothing or some 
fuzzy/fuggy/unclarified/undistinguished reaction.

Any help is appreciated as I am new in fish field,
Naira

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[Histonet] zebrafish

2011-08-25 Thread Bruijntjes, J.P. (Joost)
Hi all

We would like to prepare cryosections of adult zebrafish (3 - 6 months old). 
Fixation is not allowed. Is anyone of you familiar with the preparation of 
cryosections (including fish bones, and spinal cord)?
I assume that especially the spinal cord will cause problems.

Joost Bruijntjes
TNO-Triskelion
Zeist
The Netherlands


TNO.NLhttp://www.tno.nl/

Joost Bruijntjes

T +31 88 866 17 38
F +31 30 694 49 86
E joost.bruijnt...@tno.triskelion.nlmailto:joost.bruijnt...@tno.triskelion.nl

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