Use of AFM for imaging and measurement of the mechanical properties of light-convertible organelles in plants.
We obtained topographic images of etioplasts and chloroplasts and measured their elasticity in a physiological buffer using an atomic force microscope (AFM) and found a possible correlation between the morphological and mechanical properties during the light conversion of etioplasts to chloroplasts. Alcian blue 8GX dye was found to be effective for immobilizing the plant organelles stably on a glass surface for AFM experiments. We employed the tapping-mode AFM with a cantilever soft enough to measure the elasticity of the organelles in a liquid solution. The best images of soft, spherical organelles were obtained using the tapping-mode AFM with oscillation at the thermal vibration frequency of the cantilever of around 3 kHz. Whereas etioplasts were found to be smooth-surfaced and stiff against compression by the AFM tip, before light conversion to chloroplasts, they became rough-surfaced and mechanically soft after exposure to light. The elasticity of etioplasts was 20 times higher than that of chloroplasts, probably reflecting changes in their inner structures.
Yamada
Takafumi
T
Laboratory of Biodynamics, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, Japan.
Arakawa
Hideo
H
Okajima
Takaharu
T
Shimada
Takayoshi
T
Ikai
Atsushi
A
D002736:Chloroplasts / Q000648:ultrastructure*; D018553:Cucumis sativus / Q000648:ultrastructure*; D004548:Elasticity; D008027:Light; D018625:Microscopy, Atomic Force / Q000295:instrumentation / Q000379:methods*; D015388:Organelles / Q000648:ultrastructure*; D018724:Spinacia oleracea / Q000648:ultrastructure*