Experimental point-projection shadow microscope (PPM) images of uncoated, unstained purple membrane (PM, bacteriorhodopsin, a membrane protein from Halobacterium holobium) were obtained recently using 100 volt electrons. The membrane thickness is about 5 nm and the hexagonal unit cell dimension 6 nm. The images show contrast around the edges of small holes, as shown in figure 1. The interior of the film is opaque. Since the inelastic mean free path for 100V electrons in carbon (about 6 Å) is much less than the sample thickness, the question arises that how much, if any, transmission of elastically scattered electrons occurs. A large inelastic contribution is also expected, attenuated by the reduced detection efficiency of the channel plate at low energies. Quantitative experiments using an energy-loss spectrometer are planned. Recently Shedd has shown that at about 100V contrast in PPM images of thin gold films can be explained as Fresnel interference effects between different pinholes in the film, separated by less than the coherence width.
Lipid-protein stoichiometries in a crystalline biological membrane: NMR quantitative analysis of the lipid extract of the purple membrane