Posted by
Sudipta Maiti on
URL: http://confocal-microscopy-list.275.s1.nabble.com/deep-UV-imaging-tp1364588p1367069.html
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