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UV Imaging

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.

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UV Imaging

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  1. UV Imaging

  2. Formulatrix UV Imaging: -Overview-Performance-Feedback-Future Developments

  3. 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 [Lysozyme: 4.7% - Typical Protein, 1.4%] • And, under ultraviolet light, tryptophan fluoresces! Absorption Emission

  4. UV Overview UV camera • Emitted by the UV light source UV Source Protein crystal containing tryptophan

  5. UV Overview • Emitted by the UV light source • Passes through excitation filter Excitation filter

  6. UV Overview UV camera • Emitted by the UV light source • Passes through excitation filter • Reflected by dichroic mirror UV-compatible optics Dichroic mirror

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

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

  9. 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

  10. UV Overview • Emitted by the UV light source • Passes through excitation filter • Reflected by dichroic mirror • Absorbed by tryptophan Emission filter • Emitted by fluorescing tryptophan • Passes through dichroic mirror • Passes through emission filter

  11. 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 • Strikes UV-sensitive camera

  12. UV Performance [ Good Looking Shot ]

  13. UV Performance [ Plate view ]

  14. UV Feedback Some 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

  15. UV Feedback Some 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

  16. UV Feedback Positive Features: - Good tool for validation - Works very well with >30um crystals - Works exceptionally well with 2D crystals [particularly Lipidic Cubic Phase]

  17. UV Feedback Positive 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

  18. UV Feedback Positive 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

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

  20. UV Development

  21. UV Development

  22. UV Development Software Projects: -Improved Automation -Autoexposure -Autofocus -Auto Region-of-Interest-Better Comparison Tools-AutoScoring

  23. UV Development [Side by side mockup] Cloned Cursor Actual Cursor

  24. UV Development [Autoscoring]

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