microscopes etc.

Paul Z. Myers myers at astro.ocis.temple.edu
Thu Oct 27 07:50:47 EST 1994


In article <01HIQFWNMAIE0034NZ at BIOMED.MED.YALE.EDU> ,
ROMAN at BIOMED.MED.YALE.EDU writes:
>Hi. I'm in the process of setting up a zebrafish colony and need some
>info on scopes etc. We hope to follow some cranial neural crest cells
>after microinjection. We would ultimately like to do some video
recording.
>I would appreciate some advice from the experts in the field as to what
>the basic scope features needed and approximate cost. Any other advise
>in terms of microinjection set ups will also be appreciated. In addition
>to my excitement in embarking into "fish" i find listening to the 
>tanks during the day. We have been successful in setting up a small
>colony in a 4 x 3 x 8 foot closet. If anyone is interested let me know.
Thanks

Hmmmm...this isn't the kind of question I can properly answer in a quick
post,
since there are a lot of variables involved and you haven't given a lot
of 
information. I can tell you how I picked my scope setup, and that might
help
you get started.

First step is to define how you're going to look at things. For zebrafish
work, DIC is generally essential. You talk about microinjection--if you
are using fluorescent tracers, you're also going to need epifluorescence.

You'll also need a range of objectives. I use a 4x or more often a 10x
for some of my cruder injection work, and a long w.d. 40x for single
cell injections. I make time-lapse observations of fine structure in 
growth cones (well, in my dreams at least), so I also have a 60x oil
immersion lens. You will want apochromats. Plan objectives are nice, but
may not be that much of an advantage in fish work, since everything seems
to be on a curved surface anyway.

If you are going to use this scope for microinjection, you'll either
want a fixed stage or a stage-mounted micromanipulator. (At least that's
the way I like to work--otherwise, I find myself absent-mindedly
fine-tuning
the focus and wanging my cells into the micropipette)

You'll also want a trinoc, for video. I've also found an opti-zoom or
optivar
is very useful for video, to help compensate for some of the inevitable
pixelation of fine structure you get with video.

Anyway, make this kind of general list of essential features, and contact
your local microscope reps. Just to give you an idea of what to expect, I
got quotes from Zeiss, Olympus, Nikon, and Leica on this kind of scope and
miscellaneous attachments. I settled on a Nikon Optiphot, which had a 
reasonable price (about $20K) and very sharp optics. A micromanipulator
was another $5K. The video gear doubled (at least) the cost of my scope--
add another $25K+ for a CCD camera and intensifier, monitors, computer,
and intensifier. If you want to make movies, storage will be another
substantial expense. An OMDR will cost you about $15K; a time-lapse VCR
(which I've found to be more nuisance than anything) about $5K; and
digital image storage will require a computer+digitizer ($5K), and lots
of storage (a big hard disk, and tape or magneto-optical archiving
device, $3-5K). And don't forget software...although, since I have a
vested interest in a certain software package, I will refrain from making
recommendations. 


------------------------------------------------------------
Paul Z. Myers                    myers at astro.ocis.temple.edu
Dept. of Biology                              (215) 204-8848
Temple University
Philadelphia, PA 19122




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