1. Douglas Instruments Microseeding slide 1 Microseeding Matrix Screening and other developments in automatic protein crystallization Patrick Shaw Stewart Douglas Instruments Limited (near Oxford, UK): Peter Baldock, Patrick Shaw Stewart, James Smith

2. Douglas Instruments Microseeding slide 2 Microseeding in screening experiments Allan D’Arcy Novartis, Basle 2006 ‘Matrix-seeding script’ An automated microseed matrix-screening method for protein crystallization. Acta Cryst. (2007). D63, 550–554

3. Douglas Instruments Microseeding slide 3 Phase diagram of a protein [Protein] [Precipitant] clear precipitate nucleation metastable zone

4. Douglas Instruments Microseeding slide 4 Microseeding in screening experiments Gregory Ireton and Barry Stoddard. 'Microseed matrix screening to improve crystals of yeast cytosine deaminase'. Acta Crystallographica section D60 (2004), 601–605. With Ca2+ conditions they couldn’t get crystals without microseeding.

5. Douglas Instruments Microseeding slide 5 Phase diagram of a protein [Protein] [Precipitant] clear precipitate metastable zone

6. Douglas Instruments Microseeding slide 6 Microseeding in screening experiments 1. May explain why there are SO many hits with MMS 2. Can be used to CONTROL number of conditions

7. Douglas Instruments Microseeding slide 7 Microseeding in screening experiments James Smith 2006 ‘Matrix-seeding script’

8. Douglas Instruments Microseeding slide 8 Microseeding in screening experiments James Smith 2006 ‘Matrix-seeding script’

9. Douglas Instruments Microseeding slide 9 Microseeding in screening experiments Lesley Haire, NIMR: 6 hits in a screen, all poorly formed. Seeding - 30 hits, including several well-formed crystals. Jens-Christian Navaro Poulsen, University of Copenhagen 1 hit in 288 wells Seeding - 10 hits in 96 wells Using one of those crystals for a second-generation seeding experiment - another 10 hits in 96 wells

10. Douglas Instruments Microseeding slide 10 Microseeding in screening experiments No seeds Laura Cendron University of Padova

11. Douglas Instruments Microseeding slide 11 Microseeding in screening experiments No seedsSeeds Laura Cendron University of Padova

12. Douglas Instruments Microseeding slide 12 Phase diagram of a protein [Protein] [Seeds]

13. Douglas Instruments Microseeding slide 13 Mini-optimization

14. Douglas Instruments Microseeding slide 14 Under £24,000 Under $46,000

15. Douglas Instruments Microseeding slide 15 Bullet points: 100 + 100 nl sitting drop Only 9.8 μl of protein to set up a 96-well plate Additive screens Microseeding Mini-optimization and optimization

16. Douglas Instruments Microseeding slide 16 Jonathan Hadden 96-well plate takes 8 minutes There is an evaporation shield

17. Douglas Instruments Microseeding slide 17 Under £24,000 Under $46,000

18. Douglas Instruments Microseeding slide 18 ORYX (Arabian) The Biblical Zoo in Jerusalem