Abstract
Fixed-target serial femtosecond crystallography (FT-SFX) was an important advance in crystallography by dramatically reducing sample consumption, while maintaining the benefits of SFX for obtaining crystal structures at room temperature without radiation damage. Despite a number of advantages, preparation of a sample holder for the sample delivery in FT-SFX with the use of many crystals in a single mount at ambient temperature is challenging as it can be complicated and costly, and thus, development of an efficient sample holder is essential. In this study, we introduced a nylon mesh-based sample holder enclosed by a polyimide film. This sample holder can be rapidly manufactured using a commercially available nylon mesh with pores of a desired size at a low cost without challenging technology. Furthermore, this simple device is highly efficient in data acquisition. We performed FT-SFX using a nylon mesh-based sample holder and collected over 130,000 images on a single sample holder using a 30 Hz X-ray pulse for 1.2 h. We determined the crystal structures of lysozyme and glucose isomerase using the nylon mesh at 1.65 and 1.75 Å, respectively. The nylon mesh exposed to X-rays produced very low levels of background scattering at 3.75 and 4.30 Å, which are negligible for data analysis. Our method provides a simple and rapid but highly efficient way to deliver samples for FT-SFX.
Highlights
The fixed-target method can be divided into two approaches, one using a multi-shot goniometer approach using larger crystals, and the other using a multiple microcrystal approach[28]
When the nylon loop is used as the FT-SFX sample holder, the crystal sample sinks downward by gravity if the holder is set up vertically for X-ray exposure
We fabricated a sample holder based on nylon mesh enclosed by a polyimide film
Summary
The fixed-target method can be divided into two approaches, one using a multi-shot goniometer approach using larger crystals, and the other using a multiple microcrystal approach[28]. The multi-shot goniometer method exposes X-rays to renew volumes each time on a large crystal sample[28]. In case of micro-grid, it is necessary to change the sample several times while collecting full data since the chip area to accommodate crystals is small[17,19] These sample holders may require suitable mounting holes each time depending on the size and shape of crystals for the stable deposition of crystals. Among various applications of nylon materials, nylon loops are widely used to mount crystal samples in single crystal X-ray diffraction experiments[34] This loop provides extremely stable support for crystal samples, and the nylon material produces insignificant X-ray background scattering[34,35]. Our results are applicable to FT-SFX and to serial diffraction data collection using a synchrotron
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