Basefree

Heh. As I was reading through this post I was thinking, "Wow, that sounds really cool, but how the hell do you raise germ-free fish?" And that's when I got to the part about it being a pain in the ass.
How do you even start to get an embryo germ-free? Do you have to autoclave the tank?

And why does is the BrdU staining so weird? (or is that normal BrdU for a zebrafish?)

Most importantly, though, which lab are you leaning towards at the moment?

What is weird about the BrdU stain? The way I am interpreting it is that the green dots are cells that were synthesizing new DNA during the 15 minutes that the larvae were soaked in the BrdU solution. Then, I fixed the poor bastards, sectioned them, and labeled the cells that were proliferating during the 15 min with anti-BrdU-fluorescein antibody.

Currently, I am leaning towards the Johnson Lab. All three PI's that I rotated with have given me their recruitment sales pitches, and I am still seriously considering working in the lab I am in now. It is a tough call. Eric has lots of cool proteomic stuff lined up that I could work on, and that is sort of what I want to get in to. However, Karen works with bacteria, and I have liked bacteria for a super long time. Both labs have fun atmospheres and plenty of funding. Karen and Eric also happen to be married, so which ever lab I choose, I will still get to go to the social functions for both labs. It is a hard decision to make.

Oh yeah, the germ-free fish... I have been putting embryos (still in their chorions) through a series of dilute bleach solutions. Supposedly the protocol has worked in the past, but I don't know if I believe it.

i remember the sad part of your thesis presentation when someone asked about the zebrafish and you explained that you killed them.

but the part your advisor did with the pictures sort of made up for that. ha ha! tom cruises' body!

flash/flesh gordon?

*looks back at my original comment*
Hmmm. Maybe if I had proofread that...what I mean to say was, "Why does the BrdU stain look so weird?" I was wondering what's up with that pattern of the cells actively replicating (or lack of pattern, as the case may be)? Maybe I'm just not orienting myself properly....is this part of a grown fish, or a developing embryo on top of the yolk sac?

Dilute bleach? No wonder you're skeptical. Has anyone tried just hitting them with all the old favorite antibiotics (kan, amp, chlor, and rif)? You could homogenize the little buggers and plate out the mix to see if you killed everything...

The picture sort of resembles the Bat-symbol, used by Gotham City Police Commisioner Gordon?

Gordon :
I assumed it was a reference to Gorton's, the huge Gloucester based fish company.

http://www.gortons.com/

-G

to the bat cave!

I don't really know what is up with the lack of a pattern of cell division. Perhaps I could explain it if I had more of a background in zebrafish development. The picture is a cross section of a 4 days post-fertilization larvae. The yolk sack is on the bottom. You can sort of see a pectoral fin on the left side of the picture. If it were an adult, you might expect to see a pattern of cell division similar to what is seen in mammals, where stem cells in the intestinal crypts continually divide, however, we don't know for sure if it works that way in zebrafish, and we are nowhere near being able to raise germ-free adults.

No gold stars have been awarded yet...here is a hint: PNAS

Hmmmm....I'm going to need a bigger hint than the name of a journal...

sounds like 'penis'?

I have decided to award gold stars to all who participated in the Gordon Guessing Game. It seems that Jeff Gordon is only well known among people who are into gnotobiology. He is pretty much THE big name among scientists that study germ-free animals. His lab published a cool paper in PNAS last month that showed that germ-free mice have less developed intestinal blood vessels than conventional mice. Exciting stuff in the lab I am working in, but probably not for the rest of the world.