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Near-Field Scanning Optical Images of Bacteria

Published online by Cambridge University Press:  17 March 2011

Ana M. de Paula ,
Juliana A. Toledo ,
Haroldo B. Silva  and
Gerald Weber
Ana M. de Paula
Affiliation:
Laboratório de Nano-Espectroscopia Óptica, Universidade São Francisco, Av. São Francisco de Assis 218, 12916-900 Bragança Paulista-SP, Brazil
Juliana A. Toledo
Affiliation:
Laboratório de Nano-Espectroscopia Óptica, Universidade São Francisco, Av. São Francisco de Assis 218, 12916-900 Bragança Paulista-SP, Brazil
Haroldo B. Silva
Affiliation:
Laboratório de Nano-Espectroscopia Óptica, Universidade São Francisco, Av. São Francisco de Assis 218, 12916-900 Bragança Paulista-SP, Brazil
Gerald Weber
Affiliation:
Laboratório de Nano-Espectroscopia Óptica, Universidade São Francisco, Av. São Francisco de Assis 218, 12916-900 Bragança Paulista-SP, Brazil
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Abstract

Near field scanning optical microscopy (NSOM) and spectroscopy techniques were used to study the shape and the cell membrane details in bacteria. We present transmission and topo-graphic images for the bacteria Pseudomonas aeruginosa using the Aurora NSOM from ThermoMicroscopes. The P. aeruginosa has been widely studied due to its clinical importance in many infectious diseases. The samples were stained by the Gram method and we measured the absorption of laser light at 488 nm by the dye (Safranin) fixed at the bacteria membrane. To obtain good images we had to improve the sample preparation in order to obtain isolated bacteria at the microscope slab. This was achieved using individual colonies, from a dry growth procedure, diluted in physiologic solution. Comparison of the topographic and transmission images give information on the shape and details of the absorption of the laser light by the cell membrane. The results show patterns that depend on the thickness and shape of the membrane, thus revealing details of the cell membrane with nanometer resolution. These results could be an important tool for instance in studies of the effects of antibiotics on the cell membrane.

Type
Research Article
Information
MRS Online Proceedings Library (OPL) , Volume 711: Symposia FF/GG/HH – Advanced Biomaterials--Characterization, Tissue Engineering and Complexity , 2001 , FF7.8.1
Copyright
Copyright © Materials Research Society 2002

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References

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