Supplementary Figure 3: coMESH injection of 30S ribosomal subunit, PSII and thermolysin crystals.

The scale bars in each picture are 360 μm. (a) 30S ribosomal subunit crystals: 17% (v/v) MPD mother liquor in 100 μm ID capillary. The sheath flow capillary is 75 μm ID,running 34% (v/v) MPD sister liquor. The outer concentric capillary is 200 μm ID × 5 cm long tapered fused silica capillary coterminal with the inner capillary. The sheath flow was charged by a charging union at 3,000 V and the counter electrode was grounded. The sample ran at 1.5 μl/min for almost an hour, hence the buildup evident from debris created by the intense X-ray interaction. (b) PSII crystals (same capillary conditions as (a)): with the appropriate PSII mother and sister liquor described. Here the sample flow was charged by the syringe reservoir’s needle to 7,000 V with the counter electrode at ground. The syringe pump was set to 1 μl/min. (c) PSII crystals in modified protruded configuration: Here the flow parameters and geometries are the same as in (b), with the exception that the outer line is simply primed and then capped off and the outer concentric capillary is recessed about 1 mm. (d) Thermolysin crystal standard in modified protruded coMESH: similar flow conditions to (c) with samples in 38% (v/v) glycerol, 50 mM CaCl₂ and 0.1M MES. The speckles in the capillary are crystals flowing towards the interaction region.