used microscope for home-building a sted (super-res) system?

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Usman Qazi Usman Qazi
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used microscope for home-building a sted (super-res) system?

To build such a system most economically, I’d like to get some opinions on component prices.

 

Primarily, I am looking for a used epifluorescence microscope that can be equipped with a piezo-electric stage. My initial web searches have not yielded much on the used equipment market.

 

But at the minimum, I’d like to salvage a suitable ‘shell’ from somewhere. The more optics there are on it, the better.

 

If anyone has some ideas to share regarding the other components needed to build a STED system, please share by all means. For example, is it better to go for a blue diode or a Ti:Sapphire laser?

 

I haven’t seen a STED system in action. But it could be useful for pathologists in a developing countries where EM’s are very costly to buy and maintain. I used to do 3DEM, but need to adapt myself to the environment here.

 

Regards,

 

Usman Qazi, PhD

School of Science and Engineering

LUMS

DHA, Lahore Cantt, Pakistan

 

sse.lums.edu.pk

 

Rietdorf, Jens Rietdorf, Jens
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Re: used microscope for home-building a sted (super-res) system?

Dear Usman,

 

It might be more simple and cheaper to build an instrument exploiting the  PALM/STORM or RESOLFT methods to increase resolution.

 

Best, jens

 

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Usman Qazi
Sent: Tuesday, March 10, 2009 02:04 AM
To: [hidden email]
Subject: used microscope for home-building a sted (super-res) system?

 

To build such a system most economically, I’d like to get some opinions on component prices.

 

Primarily, I am looking for a used epifluorescence microscope that can be equipped with a piezo-electric stage. My initial web searches have not yielded much on the used equipment market.

 

But at the minimum, I’d like to salvage a suitable ‘shell’ from somewhere. The more optics there are on it, the better.

 

If anyone has some ideas to share regarding the other components needed to build a STED system, please share by all means. For example, is it better to go for a blue diode or a Ti:Sapphire laser?

 

I haven’t seen a STED system in action. But it could be useful for pathologists in a developing countries where EM’s are very costly to buy and maintain. I used to do 3DEM, but need to adapt myself to the environment here.

 

Regards,

 

Usman Qazi, PhD

School of Science and Engineering

LUMS

DHA, Lahore Cantt, Pakistan

 

sse.lums.edu.pk

 

Guy Cox Guy Cox
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Re: used microscope for home-building a sted (super-res) system?

In reply to this post by Usman Qazi
Lets be clear from the start that STED microscopes are likely to be more costly and more difficult to maintain than electron microscopes.  EM technology is mature, low-end systems are cheap, and maintenance is straightforward.  Alignment accuracy required for a STED system is probably 2 orders of magnitude higher than alignment of a TEM.

Having said that, it's also true that starting from nothing you probably have more chance of building a useful homebrew STED system than you do of building a homebrew TEM.  Your first decision is whether to build a system like the Leica, which offers super-resolution only in XY, or to follow Stefan Hell's original scheme and seek super-resolution in all 3 dimensions (but markedly better in Z).  This all comes down to the apodization scheme for the depletion spot.  I'd vote for the Leica way, but their system is not the only way to do it.  I think, for example, you could get a 'doughnut' spot with a conical lens. Tony Wilson, at Oxford, could probably advise on this.

You also ask about lasers.  Again, to get a robust system your depletion beam needs to be orders of magnitude stronger than your excitation beam.  That, of course, is what made Leica go to a Ti-S for their depletion beam (plus the fact that, living as they do in the commercial world, it means you can immediately use the same microscope for two-photon imaging).  But Ti-S lasers are very expensive, and if you went to shorter wavelengths you could get equivalent resolution with a smaller difference between the excitation and depletion beam.  You will need to get your suppliers to give you the required synchonizations of the pulses.

It's all doable.  But I'd have to say my advice would be don't.  If you want to get a cheap way into super-resolution then contact Trevor Smith at Melbourne University who has developed a very clever and simple structured illumination system (based on Mats Gustafsson's research) using an everyday video projector as the illumination source.  It won't match STED in resolution, but it will go well beyond Rayleigh and, most importantly for you, it's relatively straightforward.

                                                Guy


Optical Imaging Techniques in Cell Biology
by Guy Cox    CRC Press / Taylor & Francis
    http://www.guycox.com/optical.htm
______________________________________________
Associate Professor Guy Cox, MA, DPhil(Oxon)
Electron Microscope Unit, Madsen Building F09,
University of Sydney, NSW 2006
______________________________________________
Phone +61 2 9351 3176     Fax +61 2 9351 7682
Mobile 0413 281 861
______________________________________________
     http://www.guycox.net <http://www.guycox.net/>  

 

________________________________

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Usman Qazi
Sent: Tuesday, 10 March 2009 12:04 PM
To: [hidden email]
Subject: used microscope for home-building a sted (super-res) system?



To build such a system most economically, I’d like to get some opinions on component prices.

 

Primarily, I am looking for a used epifluorescence microscope that can be equipped with a piezo-electric stage. My initial web searches have not yielded much on the used equipment market.

 

But at the minimum, I’d like to salvage a suitable ‘shell’ from somewhere. The more optics there are on it, the better.

 

If anyone has some ideas to share regarding the other components needed to build a STED system, please share by all means. For example, is it better to go for a blue diode or a Ti:Sapphire laser?

 

I haven’t seen a STED system in action. But it could be useful for pathologists in a developing countries where EM’s are very costly to buy and maintain. I used to do 3DEM, but need to adapt myself to the environment here.

 

Regards,

 

Usman Qazi, PhD

School of Science and Engineering

LUMS

DHA, Lahore Cantt, Pakistan

 

sse.lums.edu.pk

 


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