AW: Keeping zebrafish embryos in the field of view but letting them grow.

[Ekaterina PAPUSHEVA-2]

Yes, that is the trick. We are using plastic molds for it: dip it into hot agarose in a petridish, and let it solidify. Then pull the mold out.
Contact me directly, i will send you photos!
Katja



----- Reply message -----
Von: "Guy Cox" <[hidden email]>
An: "[hidden email]" <[hidden email]>
Betreff: Keeping zebrafish embryos in the field of view but letting them grow.
Datum: Do., Jun 5, 2014 04:20

Shalin,

                I know they do this sort of thing in the Monash Micro-Imaging Live Cell course so maybe one of the Steves will reply.  My own suggestion would to punch a hole just smaller than the FOV in your agarose, so the embryo can grow in any direction without you losing track of it.  A cow-sized hypodermic needle would probably do it.  (I've got several but I think it would be easier if you sourced them locally).

                                Guy

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Shalin Mehta
Sent: Thursday, 5 June 2014 6:15 AM
To: [hidden email]
Subject: Keeping zebrafish embryos in the field of view but letting them grow.

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Hello listers,
We are imaging zebrafish embryos with 10x and the embryos just about fit the field of view at the development stage of interest (4hpf-30hpf).

Are there tried and tested methods for giving the embryos room to grow, while keeping them within the field of view?

Embryos are mounted in agarose and we can make space in agarose pad near the tail when the embryos are older. But, identifying the growth axis at young stages (~4hpf) is difficult.

All inputs appreciated,
Thanks

Assistant Research Scientist,
Marine Biological Laboratory,
7 MBL Street, Woods Hole MA 02543, USA

website: http://mshalin.com
(office) Lillie 110, (ph) 508-289-7374.

[Theresa Richardson-Metzger]

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There is a product on the market that might fit your needs.  It is a standard slide format with excellent optical properties for microscopy.  It is a cell culture product called μ-Slide Angiogenesis which was designed to image cells on gel matrices, e.g. agarose*, collagen gels, hyaluronic gels, or Matrigel or directly on the coverslip-like plastic bottom.  It requires on 10ul of gel per its 4mm well.  

Here is the link:  http://ibidi.com/xtproducts/en/ibidi-Labware/Open-Slides-Dishes:-ibidi-Standard-Bottom/m-Slide-Angiogenesis

--Theresa


-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Ekaterina PAPUSHEVA
Sent: Thursday, June 05, 2014 1:29 AM
To: [hidden email]
Subject: AW: Keeping zebrafish embryos in the field of view but letting them grow.

Yes, that is the trick. We are using plastic molds for it: dip it into hot agarose in a petridish, and let it solidify. Then pull the mold out.
Contact me directly, i will send you photos!
Katja



----- Reply message -----
Von: "Guy Cox" <[hidden email]>
An: "[hidden email]" <[hidden email]>
Betreff: Keeping zebrafish embryos in the field of view but letting them grow.
Datum: Do., Jun 5, 2014 04:20

Shalin,

                I know they do this sort of thing in the Monash Micro-Imaging Live Cell course so maybe one of the Steves will reply.  My own suggestion would to punch a hole just smaller than the FOV in your agarose, so the embryo can grow in any direction without you losing track of it.  A cow-sized hypodermic needle would probably do it.  (I've got several but I think it would be easier if you sourced them locally).

                                Guy

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Shalin Mehta
Sent: Thursday, 5 June 2014 6:15 AM
To: [hidden email]
Subject: Keeping zebrafish embryos in the field of view but letting them grow.

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Hello listers,
We are imaging zebrafish embryos with 10x and the embryos just about fit the field of view at the development stage of interest (4hpf-30hpf).

Are there tried and tested methods for giving the embryos room to grow, while keeping them within the field of view?

Embryos are mounted in agarose and we can make space in agarose pad near the tail when the embryos are older. But, identifying the growth axis at young stages (~4hpf) is difficult.

All inputs appreciated,
Thanks

Assistant Research Scientist,
Marine Biological Laboratory,
7 MBL Street, Woods Hole MA 02543, USA

website: http://mshalin.com
(office) Lillie 110, (ph) 508-289-7374.