Microcrystal Preparation
Microcrystal Preparation - Related Videos
09:09
Preparation and Delivery of Protein Microcrystals in Lipidic Cubic Phase for Serial Femtosecond Crystallography
0 Views •
Cited by 20 •
2016
We describe procedures for the preparation and delivery of membrane protein microcrystals in lipidic cubic phase for serial crystallography at X-ray free-electron lasers and synchrotron sources. These protocols can also be applied for incorporation and delivery of soluble protein microcrystals, leading to substantially reduced sample consumption compared to liquid injection.
09:00
A Sample Preparation Pipeline for Microcrystals at the VMXm Beamline
0 Views •
Cited by 10 •
2021
The signal-to-noise ratio of data is one of the most important considerations in performing X-ray diffraction measurements from microcrystals. The VMXm beamline provides a low-noise environment and microbeam for such experiments. Here, we describe sample preparation methods for mounting and cooling microcrystals for VMXm and other microfocus macromolecular crystallography beamlines.
09:48
Microcrystal Electron Diffraction of Small Molecules
0 Views •
Cited by 8 •
2021
Here, we describe the procedures developed in our laboratory for preparing powders of small molecule crystals for microcrystal electron diffraction (MicroED) experiments.
07:26
Improving High Viscosity Extrusion of Microcrystals for Time-resolved Serial Femtosecond Crystallography at X-ray Lasers
0 Views •
Cited by 27 •
2019
The success of a time-resolved serial femtosecond crystallography experiment is dependent on efficient sample delivery. Here, we describe protocols to optimize the extrusion of bacteriorhodopsin microcrystals from a high viscosity micro-extrusion injector. The methodology relies on sample homogenization with a novel three-way coupler and visualization with a high-speed camera.
03:00
Preparation of Epoxides
0 Views •
2023
Overview Epoxides result from alkene oxidation, which can be achieved by a) air, b) peroxy acids, c) hypochlorous acids, and d) halohydrin cyclization. Epoxidation with Peroxy Acids Epoxidation of alkenes via oxidation with peroxy acids involves the conversion of a carbon–carbon double bond to an epoxide using the oxidizing agent meta-chloroperoxybenzoic acid, commonly known as MCPBA. Since the O–O bond of peroxy acids is very weak, the addition of electrophilic oxygen of peroxy acids to...
FAQs
Supersaturation provides the thermodynamic driving force for molecules to leave solution and assemble into an ordered crystal lattice. Reaching this state promotes nucleation, the formation of initial crystal seeds, while carefully limiting the available growth conditions helps keep the resulting crystals small. Controlling this balance is important when uniform microcrystals are needed for downstream structural experiments.
Precipitant concentration, pH, temperature, and solvent composition determine whether nucleation and growth occur efficiently. Changing any of these conditions can alter the degree of supersaturation and therefore the number and size of crystals produced. Systematic adjustment helps researchers identify conditions that favor many suitably small crystals rather than uncontrolled growth or inadequate crystal formation.
Microcrystals can provide a practical alternative when a purified protein does not readily form large crystals. Their small size supports diffraction approaches designed to analyze many crystals, including serial femtosecond crystallography. This expands structural studies to samples whose crystal growth is limited, helping investigators examine molecular structure and relate it to biochemical function.
Screening compares crystallization conditions that differ in precipitant concentration, pH, temperature, or solvent composition. Researchers use the resulting observations to identify combinations that produce suitable nucleation and controlled growth. This optimization step is essential because the best conditions for generating microcrystals depend on the behavior of the particular purified compound or protein under study.
A typical workflow begins with a purified compound, followed by adjustment of crystallization conditions to create supersaturation. Researchers then apply a microbatch or vapor-diffusion approach and monitor the resulting crystal formation. Conditions are refined through screening until the preparation yields uniform microcrystals that can be handled for structural or diffraction experiments.
Microcrystal preparation supports structural analysis of proteins and other purified compounds by producing samples suitable for diffraction-based measurements. The resulting structural information can clarify molecular organization and support interpretation of biochemical function. In particular, these preparations enable X-ray crystallography and serial femtosecond crystallography, linking experimental structure with questions about how biomolecules operate.