Nylon mesh-based sample holder for fixed-target serial femtosecond crystallography

Area of Science:

• Crystallography
• Structural Biology
• Biophysics

Background:

• Fixed-target serial femtosecond crystallography (FT-SFX) offers room-temperature structure determination with reduced sample needs.
• Current sample holder preparation for FT-SFX is complex, costly, and hinders efficient data acquisition.
• Developing an efficient and accessible sample delivery system is crucial for advancing FT-SFX.

Purpose of the Study:

• To introduce a novel, cost-effective, and user-friendly sample holder for FT-SFX.
• To demonstrate the efficiency of this new sample holder in high-throughput data collection.
• To validate the utility of the sample holder for determining crystal structures.

Main Methods:

• A nylon mesh-based sample holder, enclosed by a polyimide film, was fabricated using commercially available materials.
• The sample holder was utilized for fixed-target serial femtosecond crystallography experiments.
• Crystal structures were determined using data collected from the novel sample holder.

Main Results:

• The nylon mesh sample holder was rapidly manufactured at low cost without complex technology.
• Over 130,000 images were collected from a single sample holder in 1.2 hours using a 30 Hz X-ray pulse.
• High-resolution crystal structures of lysozyme (1.65 Å) and glucose isomerase (1.75 Å) were determined.
• Negligible background scattering was observed from the nylon mesh upon X-ray exposure.

Conclusions:

• The developed nylon mesh sample holder provides a simple, rapid, and highly efficient method for sample delivery in FT-SFX.
• This innovation significantly reduces the complexity and cost associated with FT-SFX sample preparation.
• The method facilitates high-throughput data collection and accurate crystal structure determination.