Zeiss TIRF Module

[Stephen Bunnell]

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Dear list members,

Does anyone on list have real-world experience with the Zeiss TIRF Module?
I'm considering buying the unit, and would like to speak to someone who has
used one themselves.

    All the best,

    -Steve



****************************************************************************
Stephen C. Bunnell, Ph.D.
Assistant Professor
Tufts University Medical School
Department of Pathology
Jaharis Bldg., Room 512
150 Harrison Ave.
Boston, MA 02111

Phone: [phone]
Fax:   [phone]
Email: [hidden email]

[lechristophe]

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I'm just finishing to setup a Zeiss TIRF microscope.

- The plain Zeiss module which includes an Ar/Kr laser is a bit outdated
compared to newer setups (eg Leica TIRF multicolor) or other options
using solid-state lasers. I was happy to get a customized solid-state
laser box with an AOTF and have it hooked it to the Zeiss microscope,
the whole thing being controlled by Metamorph software. I have to say
that Zeiss and Nikon were the two companies that were really trying to
meet to my needs and not just selling me a complete turn-key setup that
was not exactly what I needed (two color 488/561nm solid state lasers +
fast 3D widefield). I've heard that Zeiss will update the laser source
in the near future to provide solid-state sources directly.

- The Zeiss slider system works quite well, but it's a manual instrument
: you have to adjust divergence and angle with manual knobs, no
motorization yet. It's a very different philosophy than say, the Leica
system where everything is controlled by the software, with an intuitive
"penetration depth" software control, but very few direct controls on
what happens inside the "black box". The overall quality of the Zeiss
TIRF illumination, once you optimize everything (laser coupling,
divergence, angles) is very good (homogeneous, stable, bright with 30 mW
lasers).


- The TIRF slider allows you to get simultaneously widefield excitation
(trough the lamp port) and TIRF laser excitation. This is done thanks to
a polarizing dichroic that puts the laser beam into the optical path. So
you can switch very rapidly between TIRF and epifluorescence (by
shuttering with a DG4/monochromator for epi and AOTF for lasers), but
the non-polarizing nature of epifluorescent light causes you to loose
50% of the epi excitation, and the slider is not easily removable if you
need full power. This power drop is not a big deal for live cell
imaging, but if you want to get single-molecule imaging or
light-demanding application, be sure to have a high-powered
epifluorescent light source (eg 300W DG4 instead of 175W).

- I don't use the Zeiss Axiovision software but I had it demoed and it's
pretty good.

- If you already have the Zeiss scope, you can go for the Zeiss TIRF
module (with the slider) or setup a customized setup using a laser
source and a Till Photonics laser coupler (different from the slider).
But in this case you will have to control everything from a dedicated
software that supports all the hardware (Metamorph, Matlab,
Slidebook...) or see with your Zeiss rep what kind of non-Zeiss hardware
is supported by Axiovision.

Hope this helps,


Christophe Leterrier






Stephen Bunnell a écrit :

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Dear list members,
>
> Does anyone on list have real-world experience with the Zeiss TIRF Module?
> I'm considering buying the unit, and would like to speak to someone who has
> used one themselves.
>
>     All the best,
>
>     -Steve
>
>
>
> ****************************************************************************
> Stephen C. Bunnell, Ph.D.
> Assistant Professor
> Tufts University Medical School
> Department of Pathology
> Jaharis Bldg., Room 512
> 150 Harrison Ave.
> Boston, MA 02111
>
> Phone: [phone]
> Fax:   [phone]
> Email: [hidden email]
>