Azim Sana, Bultema Lindsey A, de Kock Michiel B, Osorio-Blanco Ernesto Rafael, Calderón Marcelo, Gonschior Josef, Leimkohl Jan-Philipp, Tellkamp Friedjof, Bücker Robert, Schulz Eike C, Keskin Sercan, de Jonge Niels, Kassier Günther H, Miller R J Dwayne
Max Planck Institute for the Structure and Dynamics of Matter, Luruper Chaussee 149, Geb. 99 (CFEL), 22761Hamburg, Germany.
Centre for Structural Systems Biology, Department of Chemistry, University of Hamburg, Notkestraße 85, 22607Hamburg, Germany.
Microsc Microanal. 2021 Feb;27(1):44-53. doi: 10.1017/S1431927620024654.
Liquid-phase transmission electron microscopy is a technique for simultaneous imaging of the structure and dynamics of specimens in a liquid environment. The conventional sample geometry consists of a liquid layer tightly sandwiched between two Si3N4 windows with a nominal spacing on the order of 0.5 μm. We describe a variation of the conventional approach, wherein the Si3N4 windows are separated by a 10-μm-thick spacer, thus providing room for gas flow inside the liquid specimen enclosure. Adjusting the pressure and flow speed of humid air inside this environmental liquid cell (ELC) creates a stable liquid layer of controllable thickness on the bottom window, thus facilitating high-resolution observations of low mass-thickness contrast objects at low electron doses. We demonstrate controllable liquid thicknesses in the range 160 ± 34 to 340 ± 71 nm resulting in corresponding edge resolutions of 0.8 ± 0.06 to 1.7 ± 0.8 nm as measured for immersed gold nanoparticles. Liquid layer thickness 40 ± 8 nm allowed imaging of low-contrast polystyrene particles. Hydration effects in the ELC have been studied using poly-N-isopropylacrylamide nanogels with a silica core. Therefore, ELC can be a suitable tool for in situ investigations of liquid specimens.