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All sourcesNASA NTRSDOE OSTI
NASA NTRS1993

Modeling of PCG fluid dynamics: Salient results

Materials processing in space-based laboratories has already yielded higher quality crystals during previous space flights, and opportunities for several fluids experiments are anticipated during the extended duration missions planned for the future. Crystal growth in space benefits not only from its reduced gravity environment but also from the absence of the hydrostatic pressure which assists certain crystal growth and refinement methods. Gravity-driven phenomena are thus reduced in strength, and a purely diffusive fluid's behavior can be attained. In addition, past materials science experiments have shown that microgravity can also help produce larger crystals. While gravity-related effects are definitely curtailed in space, they are nevertheless present to some degree due to the acceleration environment onboard the spacecraft. This residual acceleration level is comprised of quasi-steady, oscillatory, and transient components, and is caused by a variety of mechanisms. For example, gravity gradient forces produce low frequency disturbances, and the operation of machinery, control thrusters, solar panels, human activity, etc. contribute to higher frequency accelerations. These disturbances are collectively referred to as g-jitter, and they can be deleterious to certain experiments where the minimization of the acceleration level is important. Advanced vibration isolation techniques can be utilized to actively filter out some of the detrimental frequencies and help in obtaining optimum results. However, the successful application of this technology requires the detailed analysis of candidate fluids experiments to gauge their response to g-jitter and to determine their acceleration sensitivities. Several crystal growth experiments in the Protein Crystal Growth (PCG) area, besides others, are expected to be carried out on future shuttle flights and on Space Station Freedom. The need for vibration isolation systems or components for microgravity science experiments can be expected to grow as experiments and available hardware becomes more complex. This technology will also find increased application as the science community develops an awareness of their specific needs relative to the environment available in manned space missions. Vibration isolation research strives to develop a microgravity environment requirement that defines tolerance limits on the allowable g-level, and provides the required technology to achieve it. This effort will assist in establishing the tolerable acceleration levels for specific fluids experiments. The primary effort is directed towards modeling PCG and the approach undertaken for this investigation is outlined. The objectives of this research are: (1) to computationally determine vibration sensitivity of protein crystal growth experiments; (2) determine if these experiments can benefit from vibration isolation techniques; and (3) provide realistic requirements for vibration isolation technology.

Ramachandran, N.↗
NASA NTRS2018

Microscopes on ISS - Present and Future Possibilities

The attached slides are for an invited talk (invitation from Ken Savins at CASIS) to be given July 19 at the "2018 Microgravity Molecular Crystal Growth Workshop". Ken Savins indicated that the protein crystal growth community would benefit from knowing more about what is available on ISS for microscopy. It is especially important that the folks growing protein crystals start to use the microscopes on ISS as diagnostic tools so that when their samples are returned to Earth and the results are unexpected, they are able to determine whether or not the return flight had an impact. In additional to preparing this presentation describing present and possible future LMM capabilities (if it is refurbished), we worked with Rachel A Ormsby (techshot) and Devin Ridgley (HNu Photonics) to add slides and a movie they provided of their capacities. Contact information for the NASA GRC Liquid Crystal Facility (LCF) will be included, and a couple of slides for LCF may be added when the contractor (James Kolibas at ZIN Technology) returns from vacation.

protein crystals↗
NASA NTRS2000

Acetylated Lysozyme as Impurity in Lysozyme Crystals: Constant Distribution Coefficient

Hen egg white lysozyme (HEWL) was acetylated to modify molecular charge keeping the molecular size and weight nearly constant. Two derivatives, A and B, more and less acetylated, respectively, were obtained, separated, purified and added to the solution from which crystals of tetragonal HEWL crystals were grown. Amounts of the A or B impurities added were 0.76, 0.38 and 0.1 milligram per millimeter while HEWL concentration were 20, 30 and 40 milligram per milliliter. The crystals grown in 18 experiments for each impurity were dissolved and quantities of A or B additives in these crystals were analyzed by cation exchange high performance liquid chromatography. All the data for each set of 18 samples with the different impurity and regular HEWL concentrations is well described by one distribution coefficient K = 2.15 plus or minus 0.13 for A and K = 3.42 plus or minus 0.25 for B. The observed independence of the distribution coefficient on both the impurity concentration and supersaturation is explained by the dilution model described in this paper. It shows that impurity adsorption and incorporation rate is proportional to the impurity concentration and that the growth rate is proportional to the crystallizing protein in solution. With the kinetic coefficient for crystallization, beta = 5.10(exp -7) centimeters per second, the frequency at which an impurity molecule near the growing interface irreversibly joins a molecular site on the crystal was found to be 3 1 per second, much higher than the average frequency for crystal molecules. For best quality protein crystals it is better to have low microheterogeneous protein impurity concentration and high supers aturation.

Thomas, B. R.↗
DOE OSTI2024

In situ counter-diffusion crystallization and long-term crystal preservation in microfluidic fixed targets for serial crystallography

Compared with batch and vapor diffusion methods, counter diffusion can generate larger and higher-quality protein crystals yielding improved diffraction data and higher-resolution structures. Typically, counter-diffusion experiments are conducted in elongated chambers, such as glass capillaries, and the crystals are either directly measured in the capillary or extracted and mounted at the X-ray beamline. Despite the advantages of counter-diffusion protein crystallization, there are few fixed-target devices that utilize counter diffusion for crystallization. In this article, different designs of user-friendly counter-diffusion chambers are presented which can be used to grow large protein crystals in a 2D polymer microfluidic fixed-target chip. Methods for rapid chip fabrication using commercially available thin-film materials such as Mylar, propylene and Kapton are also detailed. Rules of thumb are provided to tune the nucleation and crystal growth to meet users' needs while minimizing sample consumption. These designs provide a reliable approach to forming large crystals and maintaining their hydration for weeks and even months. This allows ample time to grow, select and preserve the best crystal batches before X-ray beam time. Importantly, the fixed-target microfluidic chip has a low background scatter and can be directly used at beamlines without any crystal handling, enabling crystal quality to be preserved. The approach is demonstrated with serial diffraction of photoactive yellow protein, yielding 1.32 Å resolution at room temperature. Fabrication of this standard microfluidic chip with commercially available thin films greatly simplifies fabrication and provides enhanced stability under vacuum. These advances will further broaden microfluidic fixed-target utilization by crystallographers.

Liu, Zhongrui↗
NASA NTRS2003

Structural Biology of Proteins of the Multi-enzyme Assembly Human Pyruvate Dehydrogenase Complex

Objectives and research challenges of this effort include: 1. Need to establish Human Pyruvate Dehydrogenase Complex protein crystals; 2. Need to test value of microgravity for improving crystal quality of Human Pyruvate Dehydrogenase Complex protein crystals; 3. Need to improve flight hardware in order to control and understand the effects of microgravity on crystallization of Human Pyruvate Dehydrogenase Complex proteins; 4. Need to integrate sets of national collaborations with the restricted and specific requirements of flight experiments; 5. Need to establish a highly controlled experiment in microgravity with a rigor not yet obtained; 6. Need to communicate both the rigor of microgravity experiments and the scientific value of results obtained from microgravity experiments to the national community; and 7. Need to advance the understanding of Human Pyruvate Dehydrogenase Complex structures so that scientific and commercial advance is identified for these proteins.

Source record↗
NASA NTRS1992

Research objectives, opportunities, and facilities for microgravity science

Microgravity Science in the U.S.A. involves research in fluids science, combustion science, materials science, biotechnology, and fundamental physics. The purpose is to achieve a thorough understanding of the effects of gravitational body forces on physical phenomena relevant to those disciplines. This includes the study of phenomena which are usually overwhelmed by the presence of gravitational body forces and, therefore, chiefly manifested when gravitational forces are weak. In the pragmatic sense, the research involves gravity level as an experimental parameter. Calendar year 1992 is a landmark year for research opportunities in low earth orbit for Microgravity Science. For the first time ever, three Spacelab flights will fly in a single year: IML-1 was launched on January 22; USML-1 was launched on June 25; and, in September, SL-J will be launched. A separate flight involving two cargo bay carriers, USMP-1, will be launched in October. From the beginning of 1993 up to and including the Space Station era (1997), nine flights involving either Spacelab or USMP carriers will be flown. This will be augmented by a number of middeck payloads and get away specials flying on various flights. All of this activity sets the stage for experimentation on Space Station Freedom. Beginning in 1997, experiments in Microgravity Science will be conducted on the Space Station. Facilities for doing experiments in protein crystal growth, solidification, and biotechnology will all be available. These will be joined by middeck-class payloads and the microgravity glove box for conducting additional experiments. In 1998, a new generation protein crystal growth facility and a facility for conducting combustion research will arrive. A fluids science facility and additional capability for conducting research in solidification, as well as an ability to handle small payloads on a quick response basis, will be added in 1999. The year 2000 will see upgrades in the protein crystal growth and fluids science facilities. From the beginning of 1997 to the fall of 1999 (the 'man-tended capability' era), there will be two or three utilization flights per year. Plans call for operations in Microgravity Science during utilization flights and between utilization flights. Experiments conducted during utilization flights will characteristically require crew interaction, short duration, and less sensitivity to perturbations in the acceleration environment. Operations between utilization flights will involve experiments that can be controlled remotely and/or can be automated. Typically, the experiments will require long times and a pristine environment. Beyond the fall of 1999 (the 'permanently-manned capability' era), some payloads will require crew interaction; others will be automated and will make use of telescience.

Bayuzick, Robert J.↗
NASA NTRS2000

Fluid Physics and Macromolecular Crystal Growth in Microgravity

The first protein crystallization experiment in microgravity was launched in April, 1981 and used Germany's Technologische Experimente unter Schwerelosigkeit (TEXUS 3) sounding rocket. The protein P-galactosidase (molecular weight 465Kda) was chosen as the sample with a liquid-liquid diffusion growth method. A sliding device brought the protein, buffer and salt solution into contact when microgravity was reached. The sounding rocket gave six minutes of microgravity time with a cine camera and schlieren optics used to monitor the experiment, a single growth cell. In microgravity a strictly laminar diffusion process was observed in contrast to the turbulent convection seen on the ground. Several single crystals, approx 100micron in length, were formed in the flight which were of inferior but of comparable visual quality to those grown on the ground over several days. A second experiment using the same protocol but with solutions cooled to -8C (kept liquid with glycerol antifreeze) again showed laminar diffusion. The science of macromolecular structural crystallography involves crystallization of the macromolecule followed by use of the crystal for X-ray diffraction experiments to determine the three dimensional structure of the macromolecule. Neutron protein crystallography is employed for elucidation of H/D exchange and for improved definition of the bound solvent (D20). The structural information enables an understanding of how the molecule functions with important potential for rational drug design, improved efficiency of industrial enzymes and agricultural chemical development. The removal of turbulent convection and sedimentation in microgravity, and the assumption that higher quality crystals will be produced, has given rise to the growing number of crystallization experiments now flown. Many experiments can be flown in a small volume with simple, largely automated, equipment - an ideal combination for a microgravity experiment. The term "protein crystal growth" is often historically used to describe these microgravity experiments. This is somewhat inaccurate as the field involves the study of many varied biological molecules including viruses, proteins, DNA, RNA and complexes of those structures. For this reason we use the term macromolecular crystal growth. In this chapter we review a series of diagnostic microgravity crystal growth experiments carried out principally using the European Space Agency (ESA) Advanced Protein Crystallization Facility (APCF). We also review related research, both experimental and theoretical, on the aspects of microgravity fluid physics that affect microgravity protein crystal growth. Our experiments have revealed some surprises that were not initially expected. We discuss them here in the context of practical lessons learnt and how to maximize the limited microgravity opportunities available.

Helliwell, John R.↗
NASA NTRS1995

STS-67 Space Shuttle mission report

The STS-67 Space Shuttle Program Mission Report provides the results of the orbiter vehicle performance evaluation during this sixty-eighth flight of the Shuttle Program, the forty-third flight since the return to flight, and the eighth flight of the Orbiter vehicle Endeavour (OV-105). In addition, the report summarizes the payload activities and the performance of the External Tank (ET), Solid Rocket Booster (SRB), Reusable Solid Rocket Motor (RSRM), and the Space Shuttle Main Engines (SSME). The serial numbers of the other elements of the flight vehicle were ET-69 for the ET; 2012, 2033, and 2031 for SSME's 1, 2, and 3, respectively; and Bl-071 for the SRB's. The left-hand RSRM was designated 360W043A, and the right-hand RSRM was designated 360L043B. The primary objective of this flight was to successfully perform the operations of the ultraviolet astronomy (ASTRO-2) payload. Secondary objectives of this flight were to complete the operations of the Protein Crystal Growth - Thermal Enclosure System (PCG-TES), the Protein Crystal Growth - Single Locker Thermal Enclosure System (PCG-STES), the Commercial Materials Dispersion Apparatus ITA Experiments (CMIX), the Shuttle Amateur Radio Experiment-2 (SAREX-2), the Middeck Active Control Experiment (MACE), and two Get-Away Special (GAS) payloads.

Fricke, Robert W., Jr.↗
NASA NTRS1993

Transport phenomena in the crystallization of lysozyme by osmotic dewatering and liquid-liquid diffusion in low gravity

Two methods of protein crystallization, osmotic dewatering and liquid-liquid diffusion, like the vapor diffusion (hanging-drop and sessile-drop) methods allow a gradual approach to supersaturation conditions. The crystallization of hen egg-white lysozyme, an extensively characterized protein crystal, in the presence of sodium chloride was used as an experimental model with which to compare these two methods in low gravity and in the laboratory. Comparisons of crystal growth rates by the two methods under the two conditions have, to date, indicated that the rate of crystal growth by osmotic dewatering is nearly the same in low gravity and on the ground, while much faster crystal growth rates can be achieved by the liquid-liquid diffusion method in low gravity.

Todd, Paul↗
NASA NTRS1994

STS-62 Space Shuttle mission report

The STS-62 Space Shuttle Program Mission Report summarizes the Payload activities as well as the Orbiter, External Tank (ET), Solid Rocket Booster (SRB), Redesigned Solid Rocket Motor (RSRM), and the Space Shuttle main engine (SSHE) systems performance during the sixty-first flight of the Space Shuttle Program and sixteenth flight of the Orbiter vehicle Columbia (OV-102). In addition to the Orbiter, the flight vehicle consisted of an ET designated as ET-62; three SSME's which were designated as serial numbers 2031, 2109, and 2029 in positions 1, 2, and 3, respectively; and two SRB's which were designated BI-064. The RSRM's that were installed in each SRB were designated as 360L036A (lightweight) for the left SRB, and 36OWO36B (welterweight) for the right SRB. This STS-62 Space Shuttle Program Mission Report fulfills the Space Shuttle Program requirement as documented in NSTS 07700, Volume 8, Appendix E. That document requires that each major organizational element supporting the Program report the results of its hardware evaluation and mission performance plus identify all related in-flight anomalies. The primary objectives of the STS-62 mission were to perform the operations of the United States Microgravity Payload-2 (USMP-2) and the Office of Aeronautics and Space Technology-2 (OAST-2) payload. The secondary objectives of this flight were to perform the operations of the Dexterous End Effector (DEE), the Shuttle Solar Backscatter Ultraviolet/A (SSBUV/A), the Limited Duration Space Environment Candidate Material Exposure (LDCE), the Advanced Protein Crystal Growth (APCG), the Physiological Systems Experiments (PSE), the Commercial Protein Crystal Growth (CPCG), the Commercial Generic Bioprocessing Apparatus (CGBA), the Middeck Zero-Gravity Dynamics Experiment (MODE), the Bioreactor Demonstration System (BDS), the Air Force Maui Optical Site Calibration Test (AMOS), and the Auroral Photography Experiment (APE-B).

Fricke, Robert W., Jr.↗
NASA NTRS2001

Effects of Convective Transport of Solute and Impurities on Defect-Causing Kinetics Instabilities

For in-situ studies of the formation and evolution of step patterns during the growth of protein crystals, we have designed and assembled an experimental setup based on Michelson interferometry with the surface of the growing protein crystal as one of the reflective surfaces. The crystallization part of the device allows optical monitoring of a face of a crystal growing at temperature stable within 0.05 C in a developed solution flow of controlled direction and speed. The reference arm of the interferometer contains a liquid-crystal element that allows controlled shifts of the phase of the interferograms. We employ an image processing algorithm which combines five images with a pi/2 phase difference between each pair of images. The images are transferred to a computer by a camera capable of capturing 6-8 frames per second. The device allows data collection data regarding growth over a relatively large area (approximately .3 sq. mm) in-situ and in real time during growth. The estimated dept resolution of the phase shifting interferometry is about 100 A. The lateral resolution, depending on the zoom ratio, varies between 0.3 and 0.6 micrometers. We have now collected quantitative results on the onset, initial stages and development of instabilities in moving step trains on vicinal crystal surfaces at varying supersaturation, position on the facet, crystal size and temperature with the proteins ferritin, apoferritin and thaumatin. Comparisons with theory, especially with the AFM results on the molecular level processes, see below, allow tests of the rational for the effects of convective flows and, as a particular case, the lack thereof, on step bunching.

Vekilov, Peter G.↗
NASA NTRS1995

STS-66 Space Shuttle mission report

The primary objective of this flight was to accomplish complementary science objectives by operating the Atmospheric Laboratory for Applications and Science-3 (ATLAS-3) and the Cryogenic Infrared Spectrometers and Telescopes for the Atmosphere-Shuttle Pallet Satellite (CRISTA-SPAS). The secondary objectives of this flight were to perform the operations of the Shuttle Solar Backscatter Ultraviolet/A (SSBUV/A) payload, the Experiment of the Sun Complementing the Atlas Payload and Education-II (ESCAPE-II) payload, the Physiological and Anatomical Rodent Experiment/National Institutes of Health Rodents (PARE/NIH-R) payload, the Protein Crystal Growth-Thermal Enclosure System (PCG-TES) payload, the Protein Crystal Growth-Single Locker Thermal Enclosure System (PCG-STES), the Space Tissue/National Institutes of Health Cells STL/N -A payload, the Space Acceleration Measurement Systems (SAMS) Experiment, and Heat Pipe Performance Experiment (HPPE) payload. The 11-day plus 2 contingency day STS-66 mission was flown as planned, with no contingency days used for weather avoidance or Orbiter contingency operations. Appendix A lists the sources of data from which this report was prepared, and Appendix B defines all acronyms and abbreviations used in the report.

Fricke, Robert W., Jr.↗
NASA NTRS2002

Dynamically controlled crystal growth system

Crystal growth can be initiated and controlled by dynamically controlled vapor diffusion or temperature change. In one aspect, the present invention uses a precisely controlled vapor diffusion approach to monitor and control protein crystal growth. The system utilizes a humidity sensor and various interfaces under computer control to effect virtually any evaporation rate from a number of different growth solutions simultaneously by means of an evaporative gas flow. A static laser light scattering sensor can be used to detect aggregation events and trigger a change in the evaporation rate for a growth solution. A control/follower configuration can be used to actively monitor one chamber and accurately control replicate chambers relative to the control chamber. In a second aspect, the invention exploits the varying solubility of proteins versus temperature to control the growth of protein crystals. This system contains miniature thermoelectric devices under microcomputer control that change temperature as needed to grow crystals of a given protein. Complex temperature ramps are possible using this approach. A static laser light scattering probe also can be used in this system as a non-invasive probe for detection of aggregation events. The automated dynamic control system provides systematic and predictable responses with regard to crystal size. These systems can be used for microgravity crystallization projects, for example in a space shuttle, and for crystallization work under terrestial conditions. The present invention is particularly useful for macromolecular crystallization, e.g. for proteins, polypeptides, nucleic acids, viruses and virus particles.

Bray, Terry L.↗
NASA NTRS1995

STS-73 Flight Day 1

On this first day of the STS-73 sixteen day mission, the crew Cmdr. Kenneth Bowersox, Pilot Kent Rominger, Payload Specialists Albert Sacco and Fred Leslie, and Mission Specialists Kathryn Thornton, Catherine 'Cady' Coleman, and Michael Lopez-Alegria are shown in various stages of prelaunch and launch activities. This mission carries the United States Microgravity Lab-2 (USML-2) payload, in which a variety of spaceborne microgravity experiments will be performed. These experiments include the Advanced Protein Crystallization Facility (APCF), The Astroculture (tm)(ASC) hardware and experiment, the Commercial Generic Bioprocessing Apparatus (CGBA), the Crystal Growth Furnace (CGF), the Drop Physics Module (DPM), the Geophysical Fluid Flow Cell (GFFC), the Glovebox (GBX), the Zeolite Crystal Growth (ZCG) experiment, the Surface Tension Driven Convection Experiment (STDCE), the Protein Crystal Growth (PCG) experiment, three Measuring Microgravity experiments (the Space Acceleration Measurement System (SAMS), the Three Dimensional Microgravity Accelerometer (3DMA), and the Orbital Acceleration Research Experiment (OARE)), and the High-Packed Digital Television (HI-PAC) demonstration system. Earth views include some cloud cover and various Earth land masses.

Source record↗
NASA NTRS1995

STS-73 Post Flight Presentation

The post flight presentation of the STS-73 Space Shuttle's United States Microgravity Lab. (USML) mission was presented by the flight crew, Cmdr. Kenneth Bowersox, Pilot Kent Rominger, Payload Specialists Albert Sacco and Fred Gregory, and Mission Specialists Kathryn Thornton, Catherine 'Cady' Collman, and Michael Lopez-Alegria, using color video and slides. Film footage includes the prelaunch and launch activities, the USML and Middeck experiments (Advanced Protein Crystallization Facility (APCF), the Astroculture(tm) (ASC) hardware and experiment, the Commercial Generic Bioprocessing Apparatus (CGBA), the Crystal Growth Furnace (CGF), the Drop Physics Module (DPM), the Geophysical Fluid Flow Cell (GFFC), the Glovebox (GBX), the Zeolite Crystal Growth (ZCG) experiment, the Surface Tension Driven Convection Experiment (STDCE), the Protein Crystal Growth (PCG) experiment, three Measuring Microgravity experiments (the Space Acceleration Measurement System (SAMS), the Three Dimensional Microgravity Accelerometer (3DMA), and the Orbital Acceleration Research Experiment (OARE)), and the High-Packed Digital Television (HI-PAC) demonstration system), pre-return flight systems checkout, reentry, and space shuttle landing. The USML experiments were monitored via the HI-PAC system downlink. Earth views included mostly geographical locations (Mediterranean Sea; Turkey; Lake Powell, Arizona/Utah area; San Francisco Bay; Baltimore, Maryland; Washington, DC; India; Tibet; China; Bhutan; Philadelphia; and the Himalayas).

Source record↗
NASA NTRS1996

STS-72 Space Shuttle Mission Report

The STS-72 Space Shuttle Program Mission Report summarizes the Payload activities as well as the Orbiter, External Tank (ET), Solid Rocket Booster (SRB), Reusable Solid Rocket Motor (RSRM), and the Space Shuttle main engine (SSME) systems performance during the seventy-fourth flight of the Space Shuttle Program, the forty-ninth flight since the return-to-flight, and the tenth flight of the Orbiter Endeavour (OV-105). In addition to the Orbiter, the flight vehicle consisted of an ET that was designated ET-75; three Block I SSME's that were designated as serial numbers 2028, 2039, and 2036 in positions 1, 2, and 3, respectively; and two SRB's that were designated BI-077. The RSRM's, designated RSRM-52, were installed in each SRB and the individual RSRM's were designated as 36OW052A for the left SRB, and 36OW052B for the right SRB. Appendix A lists the sources of data, both formal and informal, that were used to prepare this report. The primary objectives of this flight were to retrieve the Japanese Space Flyer Unit (JSFU) and deploy and retrieve the Office of Aeronautics and Space Technology-Flyer (OAST-Flyer). Secondary objectives were to perform the operations of the Shuttle Solar Backscatter Ultraviolet (SSBUV/A) experiment, Shuttle Laser Altimeter (SLA)/get-Away Special (GAS) payload, Physiological and Anatomical Rodent Experiment/National Institutes of Health-Cells (STL/NIH-C) experiment, Protein Crystal Growth-Single Locker Thermal Enclosure System (PCG-STES) experiment, Commercial Protein Crystal Growth (CPCG) payload and perform two extravehicular activities (EVA's) to demonstrate International Space Station Alpha (ISSA) assembly techniques). Appendix B provides the definition of acronyms and abbreviations used throughout the report. All times during the flight are given in Greenwich mean time (GMT) and mission elapsed time (MET).

Fricke, Robert W., Jr.↗
DOE OSTI2022

Data collection from crystals grown in microfluidic droplets

Protein crystals grown in microfluidic droplets have been shown to be an effective and robust platform for storage, transport and serial crystallography data collection with a minimal impact on diffraction quality. Single macromolecular microcrystals grown in nanolitre-sized droplets allow the very efficient use of protein samples and can produce large quantities of high-quality samples for data collection. However, there are challenges not only in growing crystals in microfluidic droplets, but also in delivering the droplets into X-ray beams, including the physical arrangement, beamline and timing constraints and ease of use. Here, the crystallization of two human gut microbial hydrolases in microfluidic droplets is described: a sample-transport and data-collection approach that is inexpensive, is convenient, requires small amounts of protein and is forgiving. It is shown that crystals can be grown in 50–500 pl droplets when the crystallization conditions are compatible with the droplet environment. Local and remote data-collection methods are described and it is shown that crystals grown in microfluidics droplets and housed as an emulsion in an Eppendorf tube can be shipped from the US to the UK using a FedEx envelope, and data can be collected successfully. Details of how crystals were delivered to the X-ray beam by depositing an emulsion of droplets onto a silicon fixed-target serial device are provided. After three months of storage at 4°C, the crystals endured and diffracted well, showing only a slight decrease in diffracting power, demonstrating a suitable way to grow crystals, and to store and collect the droplets with crystals for data collection. This sample-delivery and data-collection strategy allows crystal droplets to be shipped and set aside until beamtime is available.

59 BASIC BIOLOGICAL SCIENCES↗