Transmission Electron Microscopy of Native Nanodiscs
Caroline Brown, Snehasish Ghosh, Kallol Gupta
Abstract
This is a protocol for conducting transmission electron microscopy of native nanodiscs to determine population size distribution and morphology.
Steps
Staining grids
Dilute nanodisc samples into single molecule range, typically around between 1:10 and 1:100 is the samples have been through size exclusion chromatography using dilution buffer (50millimolar (mM) .
Glow-discharge carbon-coated copper grids (200 mesh) for 0h 0m 30s seconds.
Apply 5µL to the grid and after 0h 1m 0s minute blot off with Whiteman ashless filter paper.
Wash grid once with 5µL for 0h 0m 5s seconds.
Stain grid with 5µL for 0h 1m 0s minute and blot dry with Whiteman ashless filter paper.
Allow to dry for 0h 30m 0s minutes to
Image Acquisition
Take micrographs using a JEOL JEM 1400PLUS electron microscope at an operating voltage of 80 kV.
