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deep UV imaging

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Kevin Braeckmans

deep UV imaging

Hi everyone,

I was wondering if any of you has any experience with deep UV fluorescence imaging? In particular I would be interested in looking at the fluorescence of tryptophan (excitation around 280 nm, emission above 350 nm). Which modifications would be necessary of a standard fluorescence microscope for imaging in this region?

Thanks for any thoughts/suggestions on this.

Kevin

Prof. Dr. Kevin Braeckmans

Lab. General Biochemistry and Physical Pharmacy

Ghent University

Harelbekestraat 72

9000 Ghent

Belgium

Tel: +32 (0)9 264.80.78

Fax: +32 (0)9 264.81.89

Alberto Diaspro

Re: deep UV imaging

quartz objectives and optics

Il giorno 22/ott/08, alle ore 18:05, Kevin Braeckmans ha scritto:

Hi everyone,

I was wondering if any of you has any experience with deep UV fluorescence imaging? In particular I would be interested in looking at the fluorescence of tryptophan (excitation around 280 nm, emission above 350 nm). Which modifications would be necessary of a standard fluorescence microscope for imaging in this region?

Thanks for any thoughts/suggestions on this.

Kevin

Prof. Dr. Kevin Braeckmans
Lab. General Biochemistry and Physical Pharmacy
Ghent University
Harelbekestraat 72
9000 Ghent
Belgium
Tel: +32 (0)9 264.80.78
Fax: +32 (0)9 264.81.89

-----------------------------------------------------

Resistere, Resistere, Resistere! Hold out, Hold out, Hold out!

-----------------------------------------------------

Alberto Diaspro

Department of Physics, University of Genoa,

Via Dodecaneso 33, 16146 Genoa, Italy

fax +39-010314218 - tel +39 0103536426/309;

email: [hidden email] - URL: www.lambs.it;

THINK EBSA2009! www.ebsa2009.org

----------------------------------------------

Umberto Fascio

Re: deep UV imaging

In reply to this post by Kevin Braeckmans

quartz objectives

At 18.05 22/10/2008, you wrote:

>Hi everyone,
>
>I was wondering if any of you has any experience with deep UV
>fluorescence imaging? In particular I would be interested in looking
>at the fluorescence of tryptophan (excitation around 280 nm,
>emission above 350 nm). Which modifications would be necessary of a
>standard fluorescence microscope for imaging in this region?
>
>Thanks for any thoughts/suggestions on this.
>
>Kevin
>
>
>
>Prof. Dr. Kevin Braeckmans
>Lab. General Biochemistry and Physical Pharmacy
>Ghent University
>Harelbekestraat 72
>9000 Ghent
>Belgium
>Tel: +32 (0)9 264.80.78
>Fax: +32 (0)9 264.81.89
>
>
>
>

Dott. Umberto Fascio
C.I.M.A.
Centro Interdipartimentale Microscopia Avanzata
Via Celoria 26 - 20133 Milano
tel: +39 0250314807/14876
fax:+39 0250314802

Mario-2

Re: deep UV imaging

In reply to this post by Kevin Braeckmans

Quartz optics or at least objectives (does anybody make these,
currently?) and coverslips (fragile and expensive).

Once tried three photon excitation with some success. Might be a
reasonable option.

Mario

--
________________________________________________________________________________
Mario M. Moronne, Ph.D.

[hidden email]
[hidden email]
[hidden email]

Alberto Diaspro

Re: deep UV imaging

well
see also the three-photon excitation paper by Sudipta Maiti on
Sceince...

Il giorno 22/ott/08, alle ore 19:29, Mario ha scritto:

> Quartz optics or at least objectives (does anybody make these,
> currently?) and coverslips (fragile and expensive).
>
> Once tried three photon excitation with some success. Might be a
> reasonable option.
>
> Mario
>
>
>> Hi everyone,
>>
>> I was wondering if any of you has any experience with deep UV
>> fluorescence imaging? In particular I would be interested in
>> looking at the fluorescence of tryptophan (excitation around 280
>> nm, emission above 350 nm). Which modifications would be necessary
>> of a standard fluorescence microscope for imaging in this region?
>>
>> Thanks for any thoughts/suggestions on this.
>>
>> Kevin
>>
>>
>>
>> Prof. Dr. Kevin Braeckmans
>> Lab. General Biochemistry and Physical Pharmacy
>> Ghent University
>> Harelbekestraat 72
>> 9000 Ghent
>> Belgium
>> Tel: +32 (0)9 264.80.78
>> Fax: +32 (0)9 264.81.89
>>
>>
>>
>>
>
>
> --
> ________________________________________________________________________________
> Mario M. Moronne, Ph.D.
>
> [hidden email]
> [hidden email]
> [hidden email]

-----------------------------------------------------
Resistere, Resistere, Resistere! Hold out, Hold out, Hold out!
-----------------------------------------------------
Alberto Diaspro
Department of Physics, University of Genoa,
Via Dodecaneso 33, 16146 Genoa, Italy
fax +39-010314218 - tel +39 0103536426/309;
email: [hidden email] - URL: www.lambs.it;

THINK EBSA2009! www.ebsa2009.org
----------------------------------------------

Armstrong, Brian

Re: deep UV imaging

In reply to this post by Mario-2

Hello, I believe that Leica and Zeiss make quartz optics for use in
their industrial instruments used for Mask Alignment. These optics are
incredibly expensive because of the quartz and because adhesives cannot
be used in their construction due to the corrosive nature of UV. I know
that researchers in protein crystallography are interested in crystal
illumination in UV and that some have built their own scopes with quartz
optics. I think Reichert-Jung made quartz optics for their industrial
scopes in the past as well. Something to keep in mind if you build your
own scope is SAFETY with regards to sending UV light to the eyepieces.
Cheers,

Brian D Armstrong PhD
Light Microscopy Core Manager
Beckman Research Institute
City of Hope
Dept of Neuroscience
1450 E Duarte Rd
Duarte, CA 91010
626-256-4673 x62872
http://www.cityofhope.org/research/support/Light-Microscopy-Digital-Imag
ing/Pages/default.aspx
-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]]
On Behalf Of Mario
Sent: Wednesday, October 22, 2008 10:29 AM
To: [hidden email]
Subject: Re: deep UV imaging

Quartz optics or at least objectives (does anybody make these,
currently?) and coverslips (fragile and expensive).

Once tried three photon excitation with some success. Might be a
reasonable option.

Mario

--
________________________________________________________________________
________
Mario M. Moronne, Ph.D.

[hidden email]
[hidden email]
[hidden email]

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Kurt Thorn

Re: deep UV imaging

In reply to this post by Kevin Braeckmans

There was a recent paper in Nature Methods that built a UV
transillumination microscope for imaging protein absorption:
http://www.nature.com/nmeth/journal/v4/n7/full/nmeth1053.html

Kurt

Kevin Braeckmans wrote:

>
> Hi everyone,
>
>
>
> I was wondering if any of you has any experience with deep UV
> fluorescence imaging? In particular I would be interested in looking
> at the fluorescence of tryptophan (excitation around 280 nm, emission
> above 350 nm). Which modifications would be necessary of a standard
> fluorescence microscope for imaging in this region?
>
>
>
> Thanks for any thoughts/suggestions on this.
>
>
>
> Kevin
>
>
>
>
>
>
>
> Prof. Dr. Kevin Braeckmans
>
> Lab. General Biochemistry and Physical Pharmacy
>
> Ghent University
>
> Harelbekestraat 72
>
> 9000 Ghent
>
> Belgium
>
> Tel: +32 (0)9 264.80.78
>
> Fax: +32 (0)9 264.81.89
>
>
>
>
>
>
>
>
>

--
Kurt Thorn, PhD
Director, Nikon Imaging Center
University of California San Francisco

UCSF MC 2140
Genentech Hall Room S252
600 16th St.
San Francisco, CA 94158-2517

http://nic.ucsf.edu
phone 415.514.9709
fax 415.514.4300

Turan Erdogan

Re: deep UV imaging

In reply to this post by Kevin Braeckmans

Kevin:

Two things. First, there is a company called CRAIC (no personal affiliation) that reconfigures standard microscope stands into UV-compatible microscopes. See, for example: http://www.microspectra.com/products/uvm-1-vis-nir-microscope/34-products/53-uvm-1-ultraviolet-microscope?56563afcc766aeae8c9d5b9b84d9674a=8cba804d502960fbac4d5fa507955c58. These are also capable of fluorescence imaging. The microscope uses UV optics (objective as well as excitation-beam-line optics and tube lens).

An additional challenge is the filters. Semrock (I AM commercially affiliated with Semrock) sells a filter set specifically for imaging tryptophan (see http://www.semrock.com/Catalog/Brightline_TRP.htm).

Hope that's helpful.

-Turan

Dr. Turan Erdogan, CTO

Semrock, Inc.

3625 Buffalo Road

Rochester, NY 14624

tel: (585) 594-7001

fax: (585) 594-7095

[hidden email]

www.semrock.com

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Kevin Braeckmans
Sent: Wednesday, October 22, 2008 12:05 PM
To: [hidden email]
Subject: deep UV imaging

Hi everyone,

Thanks for any thoughts/suggestions on this.

Kevin

Prof. Dr. Kevin Braeckmans

Lab. General Biochemistry and Physical Pharmacy

Ghent University

Harelbekestraat 72

9000 Ghent

Belgium

Tel: +32 (0)9 264.80.78

Fax: +32 (0)9 264.81.89

Sudipta Maiti

Re: deep UV imaging

In reply to this post by Mario-2

We have done some UV microscopy of Tryptophan and tyrosine-like
molecules using multiphoton excitation without using quartz optics.
There are two ways to go, both of which work fairly well:
(1) for tryptophan, some objectives will pass most of it's emission at
330nm (and of course pass the three-photon excitation at ~750nm). We
have imaged serotonin quite successfully over the years using this scheme.
For recent results in various systems see
Kaushalya et al., J Neuroscience Res. vol. 86 (2008) 3469 (for neuronal cells),
Kumar et al., Stem Cells Dev. (2008) PMID 18800863 (for stem cells),
Kaushalya et al., NeuroReport vol. 19 (2008) 717 (for brain slices),
Basu et al., Reproduction, vol. 135 (2008) 657 (for embryos)
(2) For deeper UV fluorophores, e.g. dopamine emitting at 305 nm, we have
used a non-epi detector, which also gives reasonable results, see
Balaji et al., Appl. Optics vol. 43 (2004) 2412

Sorry, that was probably more advertisement than I intended.

Sudipta

> Quartz optics or at least objectives (does anybody make these, currently?)
> and coverslips (fragile and expensive).
>
> Once tried three photon excitation with some success. Might be a reasonable
> option.
>
> Mario
>
>
>> Hi everyone,
>>
>> I was wondering if any of you has any experience with deep UV fluorescence
>> imaging? In particular I would be interested in looking at the fluorescence
>> of tryptophan (excitation around 280 nm, emission above 350 nm). Which
>> modifications would be necessary of a standard fluorescence microscope for
>> imaging in this region?
>>
>> Thanks for any thoughts/suggestions on this.
>>
>> Kevin
>>
>>
>>
>> Prof. Dr. Kevin Braeckmans
>> Lab. General Biochemistry and Physical Pharmacy
>> Ghent University
>> Harelbekestraat 72
>> 9000 Ghent
>> Belgium
>> Tel: +32 (0)9 264.80.78
>> Fax: +32 (0)9 264.81.89
>>
>>
>>
>>
>
>
>

--
Sudipta Maiti
Dept. of Chemical Sciences
Tata Institute of Fundamental Research
Homi Bhabha Road, Colaba, Mumbai 400005, India
91-22-2278-2716
www.tifr.res.in/~biophotonics

Kevin Braeckmans

Re: deep UV imaging

In reply to this post by Turan Erdogan

Thanks to all who have responded, much appreciated!

I will go in more detail through the articles and other information that was suggested. For now I will keep in mind that it is not impossible, but that it will require a dedicated set-up (and budget). Triple-photon imaging is not an option in our case since the project would involve fast widefield imaging (>50 fps).

Thanks again and best regards,

Kevin

Van: Confocal Microscopy List [mailto:[hidden email]] Namens Turan Erdogan
Verzonden: woensdag 22 oktober 2008 21:16
Aan: [hidden email]
Onderwerp: Re: deep UV imaging

Kevin:

Hope that's helpful.

-Turan

Dr. Turan Erdogan, CTO

Semrock, Inc.

3625 Buffalo Road

Rochester, NY 14624

tel: (585) 594-7001

fax: (585) 594-7095

[hidden email]

www.semrock.com

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Kevin Braeckmans
Sent: Wednesday, October 22, 2008 12:05 PM
To: [hidden email]
Subject: deep UV imaging

Hi everyone,

Thanks for any thoughts/suggestions on this.

Kevin

Prof. Dr. Kevin Braeckmans

Lab. General Biochemistry and Physical Pharmacy

Ghent University

Harelbekestraat 72

9000 Ghent

Belgium

Tel: +32 (0)9 264.80.78

Fax: +32 (0)9 264.81.89

Craig Brideau

Re: deep UV imaging

In reply to this post by Turan Erdogan

Don't know if this is useful but Thorlabs sells UV objectives:

http://www.thorlabs.com/NewGroupPage9.cfm?ObjectGroup_ID=3271

No commercial interest, just use a lot of Thor stuff.

Craig

On Mon, Oct 27, 2008 at 1:11 AM, Kevin Braeckmans <[hidden email]> wrote:

Thanks to all who have responded, much appreciated!

I will go in more detail through the articles and other information that was suggested. For now I will keep in mind that it is not impossible, but that it will require a dedicated set-up (and budget). Triple-photon imaging is not an option in our case since the project would involve fast widefield imaging (>50 fps).

Thanks again and best regards,

Kevin

Van: Confocal Microscopy List [mailto:[hidden email]] Namens Turan Erdogan
Verzonden: woensdag 22 oktober 2008 21:16
Aan: [hidden email]
Onderwerp: Re: deep UV imaging

Kevin:

Two things. First, there is a company called CRAIC (no personal affiliation) that reconfigures standard microscope stands into UV-compatible microscopes. See, for example: http://www.microspectra.com/products/uvm-1-vis-nir-microscope/34-products/53-uvm-1-ultraviolet-microscope?56563afcc766aeae8c9d5b9b84d9674a=8cba804d502960fbac4d5fa507955c58. These are also capable of fluorescence imaging. The microscope uses UV optics (objective as well as excitation-beam-line optics and tube lens).

An additional challenge is the filters. Semrock (I AM commercially affiliated with Semrock) sells a filter set specifically for imaging tryptophan (see http://www.semrock.com/Catalog/Brightline_TRP.htm).

Hope that's helpful.

-Turan

Dr. Turan Erdogan, CTO

Semrock, Inc.

3625 Buffalo Road

Rochester, NY 14624

tel: (585) 594-7001

fax: (585) 594-7095

[hidden email]

www.semrock.com

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Kevin Braeckmans
Sent: Wednesday, October 22, 2008 12:05 PM
To: [hidden email]
Subject: deep UV imaging

Hi everyone,

I was wondering if any of you has any experience with deep UV fluorescence imaging? In particular I would be interested in looking at the fluorescence of tryptophan (excitation around 280 nm, emission above 350 nm). Which modifications would be necessary of a standard fluorescence microscope for imaging in this region?

Thanks for any thoughts/suggestions on this.

Kevin

Prof. Dr. Kevin Braeckmans

Lab. General Biochemistry and Physical Pharmacy

Ghent University

Harelbekestraat 72

9000 Ghent

Belgium

Tel: +32 (0)9 264.80.78

Fax: +32 (0)9 264.81.89