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![Page 1: Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission. UV Imaging](https://reader035.vdocuments.mx/reader035/viewer/2022070415/56649d385503460f94a11793/html5/thumbnails/1.jpg)
Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission.
UV Imaging
![Page 2: Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission. UV Imaging](https://reader035.vdocuments.mx/reader035/viewer/2022070415/56649d385503460f94a11793/html5/thumbnails/2.jpg)
Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission.
Formulatrix UV Imaging:
-Overview-Performance-Feedback-Future Developments
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Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission.
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
![Page 4: Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission. UV Imaging](https://reader035.vdocuments.mx/reader035/viewer/2022070415/56649d385503460f94a11793/html5/thumbnails/4.jpg)
Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission.
UV Overview Emitted by the UV light source
UV Source
Protein crystal containing tryptophan
UV camera
![Page 5: Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission. UV Imaging](https://reader035.vdocuments.mx/reader035/viewer/2022070415/56649d385503460f94a11793/html5/thumbnails/5.jpg)
Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission.
UV Overview Emitted by the UV light source Passes through excitation filter
Excitation filter
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Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission.
UV Overview Emitted by the UV light source Passes through excitation filter Reflected by dichroic mirror
UV-compatible optics
Dichroic mirror
UV camera
![Page 7: Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission. UV Imaging](https://reader035.vdocuments.mx/reader035/viewer/2022070415/56649d385503460f94a11793/html5/thumbnails/7.jpg)
Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission.
UV Overview Emitted by the UV light source Passes through excitation filter Reflected by dichroic mirror Absorbed by tryptophan
![Page 8: Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission. UV Imaging](https://reader035.vdocuments.mx/reader035/viewer/2022070415/56649d385503460f94a11793/html5/thumbnails/8.jpg)
Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission.
UV Overview Emitted by the UV light source Passes through excitation filter Reflected by dichroic mirror Absorbed by tryptophan Emitted by fluorescing tryptophan
![Page 9: Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission. UV Imaging](https://reader035.vdocuments.mx/reader035/viewer/2022070415/56649d385503460f94a11793/html5/thumbnails/9.jpg)
Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission.
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
![Page 10: Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission. UV Imaging](https://reader035.vdocuments.mx/reader035/viewer/2022070415/56649d385503460f94a11793/html5/thumbnails/10.jpg)
Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission.
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
![Page 11: Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission. UV Imaging](https://reader035.vdocuments.mx/reader035/viewer/2022070415/56649d385503460f94a11793/html5/thumbnails/11.jpg)
Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission.
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
Strikes UV-sensitive camera
Passes through emission filter
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UV Performance
[ Good Looking Shot ]
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Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission.
UV Performance
[ Plate view ]
![Page 14: Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission. UV Imaging](https://reader035.vdocuments.mx/reader035/viewer/2022070415/56649d385503460f94a11793/html5/thumbnails/14.jpg)
Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission.
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
![Page 15: Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission. UV Imaging](https://reader035.vdocuments.mx/reader035/viewer/2022070415/56649d385503460f94a11793/html5/thumbnails/15.jpg)
Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission.
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
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UV FeedbackPositive Features:
- Good tool for validation- Works very well with >30um crystals- Works exceptionally well with 2D crystals
[particularly Lipidic Cubic Phase]
![Page 17: Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission. UV Imaging](https://reader035.vdocuments.mx/reader035/viewer/2022070415/56649d385503460f94a11793/html5/thumbnails/17.jpg)
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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
![Page 18: Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission. UV Imaging](https://reader035.vdocuments.mx/reader035/viewer/2022070415/56649d385503460f94a11793/html5/thumbnails/18.jpg)
Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission.
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
Unexpected Phenomena-Florescence of aromatic compounds-Quenching of fluoresence by certain co-factors
![Page 19: Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission. UV Imaging](https://reader035.vdocuments.mx/reader035/viewer/2022070415/56649d385503460f94a11793/html5/thumbnails/19.jpg)
Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission.
UV Development
Hardware Projects:- Wider Field of view- Improved Mechanical Repeatability- Glancing Illumination- Uniform Illumination- Improved Image Sharpness
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UV Development
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UV Development
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Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission.
UV DevelopmentSoftware Projects:-Improved Automation
-Autoexposure-Autofocus-Auto Region-of-Interest
-Better Comparison Tools-AutoScoring
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UV Development
[Side by side mockup]
Actual Cursor Cloned Cursor
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UV Development
[Autoscoring]
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UV Development
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UV Development
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Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission.
UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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UV Development
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Copyright © 2009 Formulatrix, Inc. Confidential. Do not distribute without prior written permission.
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