Preparation of Oriented, Fully Hydrated Lipid Samples for Structure Determination Using X-Ray Scatte
This chapter describes a method of sample preparation called “the rock and roll method,” which is basically a solvent evaporation technique with controlled manual sample movement during evaporation of solvent from lipid/solvent mixtures that produces well-oriented thick stacks of about 2000 lipid bilayers. Many lipid types have been oriented using different solvent mixtures that balance solubilization of the lipid with uniform deposition of the lipid solution onto solid substrates. These well-oriented thick stacks are then ideal samples for collection of both X-ray diffraction data in the gel phase and X-ray diffuse scattering data in the fluid phase of lipids. The degree of orientation is determined using visual inspection, polarizing microscopy, and a mosaic spread X-ray experiment. Atomic force microscopy is used to compare samples prepared using the rock and roll method with those prepared by spin-coating, which produces well-oriented but less homogeneous lipid stacks. These samples can be fully hydrated through the vapor provided that the hydration chamber has excellent temperature and humidity control.
- 上一篇
Production of Pseudotyped Retrovirus and the Generation of Proviral Transgenic Zebrafish
This chapter describes a method for generation of the high-t...
- 下一篇
Detection of mRNA in Whole Mounts of Mouse Embryos Using Digoxigenin Riboprobes
This chapter describes a nonradioactive method for the local...