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UV Imaging

Formulatrix UV Imaging:

-Overview-Performance-Feedback-Future Developments

UV Overview Both the target protein and undesired salts will crystallize Both crystals look fundamentally similar under visible light Fortunately, only the protein crystal contains tryptophan

And, under ultraviolet light, tryptophan fluoresces!

[Lysozyme: 4.7% - Typical Protein, 1.4%]

Absorption

Emission

UV Overview Emitted by the UV light source

UV Source

Protein crystal containing tryptophan

UV camera

UV Overview Emitted by the UV light source Passes through excitation filter

Excitation filter

UV Overview Emitted by the UV light source Passes through excitation filter Reflected by dichroic mirror

UV-compatible optics

Dichroic mirror

UV camera

UV Overview Emitted by the UV light source Passes through excitation filter Reflected by dichroic mirror Absorbed by tryptophan

UV Overview Emitted by the UV light source Passes through excitation filter Reflected by dichroic mirror Absorbed by tryptophan Emitted by fluorescing tryptophan

UV Overview Emitted by the UV light source Passes through excitation filter Reflected by dichroic mirror Absorbed by tryptophan Emitted by fluorescing tryptophan Passes through dichroic mirror

Passes through emission filter

Emission filter

Strikes UV-sensitive camera

Passes through emission filter

UV Performance

[ Good Looking Shot ]

UV Performance

[ Plate view ]

UV FeedbackSome customers are happy -

“The analysis is very easy and UV imaging is now a powerful tool we are using to drive our crystallization strategies.”

- University of Marseilles

UV FeedbackSome customers are happy -

“The analysis is very easy and UV imaging is now a powerful tool we are using to drive our crystallization strategies.”

- University of Marseilles

Others are not -

“My impression is that this feature is a great idea and it will be a wonderful option when you get it to work in the future [but] The UV imaging is not part of anyone's regular workflow--we do not use it regularly since it doesn't work.”

- MPI-Heidelberg

UV FeedbackPositive Features:

- Good tool for validation- Works very well with >30um crystals- Works exceptionally well with 2D crystals

[particularly Lipidic Cubic Phase]

Common complaints:-Field of view is too tight-Image quality needs to be sharper

[Especially to resolve smaller crystals at widest field of view]-Workflow is cumbersome

[Comparing images is particularly difficult]-Requires particular crystallography media

Common complaints:-Field of view is too tight-Image quality needs to be sharper

[Especially to resolve smaller crystals at widest field of view]-Workflow is cumbersome

[Comparing images is particularly difficult]-Requires particular crystallography media

Unexpected Phenomena-Florescence of aromatic compounds-Quenching of fluoresence by certain co-factors

UV Development

Hardware Projects:- Wider Field of view- Improved Mechanical Repeatability- Glancing Illumination- Uniform Illumination- Improved Image Sharpness

UV Development

UV DevelopmentSoftware Projects:-Improved Automation

-Autoexposure-Autofocus-Auto Region-of-Interest

-Better Comparison Tools-AutoScoring

UV Development

[Side by side mockup]

Actual Cursor Cloned Cursor

UV Development

[Autoscoring]

UV Development

Questions?

copyright © 2009 formulatrix, inc. confidential. do not distribute without prior written...

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