Backscattered electron imaging and elemental analysis of rapidly frozen plant cells using variable accelerating voltage.

Yasuko Kaneko; Makoto Tokunaga; Kyoko Tanaka; Kimie Atsuzawa; Masako Nishimura

Rapidly frozen rosemary leaves were observed at variable accelerating voltages in a low-vacuum scanning electron microscope equipped with a cryo transfer system. After water was sublimated from the fractured face of the leaf, distinct backscattered electron (BSE) images were obtained depending on the accelerating voltages applied. At 5 kV, surface cell wall structure was observed, whereas at 10 and 15 kV chloroplasts lining the inside of the cell wall and membrane were visualized. With energy dispersive X-ray microanalysis, elemental information corresponding to the BSE images was obtained. Besides visualization of the structures and elemental composition close to the living state, information on layers at different depths from the surface could be detected by varying the accelerating voltage in this system.

ELECTRON backscattering; CRYOPRESERVATION of plant cells & tissues; ACCELERATING voltage scan; ROSEMARY; SCANNING electron microscopy; CHLOROPLASTS

Microscopy, 2018, Vol 67, Issue 2, p125

2050-5698

Academic Journal

10.1093/jmicro/dfx133