Has anybody any experience in using the new Zeiss LSM 700?

[Raman Rajagopal]

Zeiss has this year come up with a new system LSM 700. They claim it is spectral – by
varying the position of the second dichroic (VSD) which is an interference filter device.
The VSD, as I understand, has the ability to dynamically reflect / transmit different
wavelength along its length thus achieving diffraction of the emission into its spectral
components with out using a prism or grating.
(http://zeiss-campus.magnet.fsu.edu/tutorials/spectralimaging/lsm700/index.html - try
varying the beam splitter position in the diagram).
           
1. How would you rate LSM 700 vis a vis other machines with respect to their inherent
ability in capturing the spectral data.
2. What would be the efficiency of VSD when compared to a prism and grating method in
spectral dispersion.
3. LSM 700 has 4 solid state laser lines coming directly into the machine. They do not
mention an AOTF, but they mention that we can control individually the laser power
intensity. Can this be done with out AOTF and how effective will this be?
4.. When compared to all other machines, LSM 700 lasers have much less power: 405 – 5
mW; 488 – 10 mW; 555 – 10 mW and 639 – 5 mW. Does this mean that 700 is more
sensitive or it is a trade off for a lesser price that they offer.

 

 
RAJAGOPAL  RAMAN
Dept. of Zoology,
University of  Delhi,
New Delhi. India.

[Anil Kundalia]

Dear Sir,

I will be interested to see the public replies as the page hyperlinked does
not give any clear message.

Warm Regards.

Anil

--------------------------------------------------
From: "Raman Rajagopal" <[hidden email]>
Sent: Friday, July 03, 2009 4:49 PM
To: <[hidden email]>
Subject: Has anybody any experience in using the new Zeiss LSM 700?

> Zeiss has this year come up with a new system LSM 700. They claim it is
> spectral - by
> varying the position of the second dichroic (VSD) which is an interference
> filter device.
> The VSD, as I understand, has the ability to dynamically reflect /
> transmit different
> wavelength along its length thus achieving diffraction of the emission
> into its spectral
> components with out using a prism or grating.
> (http://zeiss-campus.magnet.fsu.edu/tutorials/spectralimaging/lsm700/index.html 
>  - try
> varying the beam splitter position in the diagram).
>
> 1. How would you rate LSM 700 vis a vis other machines with respect to
> their inherent
> ability in capturing the spectral data.
> 2. What would be the efficiency of VSD when compared to a prism and
> grating method in
> spectral dispersion.
> 3. LSM 700 has 4 solid state laser lines coming directly into the machine.
> They do not
> mention an AOTF, but they mention that we can control individually the
> laser power
> intensity. Can this be done with out AOTF and how effective will this be?
> 4.. When compared to all other machines, LSM 700 lasers have much less
> power: 405 - 5
> mW; 488 - 10 mW; 555 - 10 mW and 639 - 5 mW. Does this mean that 700 is
> more
> sensitive or it is a trade off for a lesser price that they offer.
>
>
>
>
> RAJAGOPAL  RAMAN
> Dept. of Zoology,
> University of  Delhi,
> New Delhi. India.
>

[Valeria Berno]

Dear all,

I am writing this in behalf of a colleague planning to create a
transgenic mouse with a "red" fluorescent protein. He will express the
protein in the brain (under a quite weak promoter).

I was a little bit confuse from the literature in comparing mcherry and
dTomato......What it would be your advice on choosing one of them?

I clearly need brightness and possibly also a specific antibody for the
protein working in IF or IHC.

Sure of your all useful advices

thanks

Valeria

[Kurt Thorn]

If you're not fusing the fluorescent protein to an endogenous cellular protein, tdtomato is probably your protein of choice as it's the brightest red fluorescent protein available.  If you are using it in a fusion context, you probably would prefer to use mcherry as tdtomato can perturb protein function and lead to aggregation.

Kurt

________________________________________
From: Confocal Microscopy List [[hidden email]] On Behalf Of Valeria Berno [[hidden email]]
Sent: Monday, July 06, 2009 1:46 AM
To: [hidden email]
Subject: mcherry or Tdtomato?

Dear all,

I am writing this in behalf of a colleague planning to create a
transgenic mouse with a "red" fluorescent protein. He will express the
protein in the brain (under a quite weak promoter).

I was a little bit confuse from the literature in comparing mcherry and
dTomato......What it would be your advice on choosing one of them?

I clearly need brightness and possibly also a specific antibody for the
protein working in IF or IHC.

Sure of your all useful advices

thanks

Valeria