Terese Bergfors

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Seeding Workshop

• Terese Bergfors
• Uppsala University
• Biomedical Center
• Uppsala SWEDEN

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1. How seeding works seeding and the phase
diagram
3
Phase diagram
supersaturation
Protein concentration
Precipitant conc.
4
Phase diagram
Protein concentration
Metastable zone
Precipitant conc.
5
Lower the concentration!
20 mg/ml
10 mg/ml
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2. How to seed
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Seeding methods

• Heterogeneous
• Homogeneous

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Heterogeneous seeding
fiber
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Seeding methods

• Heterogeneous
• Homogeneous
• Streak seeding
• Dilution aliquots

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Streak seeding
Enrico Stura
0.3 mm
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The seed transfer tool
Cat whisker Horse tail Acupuncture
needle 50-100 microns
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Streak dilutions
3
1
10
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Seeding advanced recipe

• Find conditions by streak seeding.
• Prepare seed stock.
• Make dilution series.
• Innoculate the new drops.

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Robotic Seeding with an Oryx
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Todays experiment Fast Lysozyme
80
60
100 mg/ml precipitant
40
20
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Streak the new drops with the parent seeds.
80
60
40
Parent crystals in 5 min
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