Electron diffraction studies of molecular ordering and orientation in phospholipid monolayer domains

Abstract

The molecular order and orientation of phase separated domains in monolayers of DP(Me)PE and DP(Me)2PE were determined by electron diffraction. Dark and bright fluorescent domains at the air-water interface were observed by fluorescence microscopy. The monolayers were transferred to Formvar coated electron microscope grids for electron diffraction studies. The positions of domains on the marker grids were recorded in fluorescence micrographs, which were used as guide maps to locate these domains in the electron microscope. Selected area electron diffraction patterns were obtained from predetermined areas within and outside the dark domains. Sharp hexagonal diffraction patterns were recorded from dark domains, and diffuse diffraction rings from bright areas in between dark domains. The diffraction results indicated that the dark domains and bright areas were comprised of lipid molecules in solid and fluid states, respectively. The orientation of diffraction patterns from adjacent locations within a dark domains changed gradually, indicating a continuous bending of the molecular packing lattice vector within these domains. Orientation directors in U-shaped DP(Me)2PE domains followed the turn of the arm; no vortex nor branching was indicated by electron diffraction. Directors branching from the "stem" of highly invaginated DP(Me)PE domains usually occurred at twinning angles of n pi/3 from the stem director, which would minimize packing defects in the development of thinner branches. Electron diffraction from local areas of individual domains proved that dark fluorescent domains were solid ones, and that pseudo-long range order existed in these solid domains.