Research Papers

Micro-resonance Raman study of optically trapped Escherichia coli cells overexpressing human neuroglobin

[+] Author Affiliations
Kerstin Ramser

Göteborgs University, Department of Physics, SE-412 96 Göteborg, Sweden and Luleå University of Technology, Department of Computer Science and Electrical Engineering, SE-971 87 Luleå, Sweden

Wim Wenseleers

University of Antwerp, Department of Physics, Wilrijk-Antwerp, Belgium

Sylvia Dewilde

University of Antwerp, Department of Biomedical Sciences, Wilrijk-Antwerp, Belgium

Sabine Van Doorslaer

University of Antwerp, Department of Physics, Wilrijk-Antwerp, Belgium

Luc Moens

University of Antwerp, Department of Biomedical Sciences, Wilrijk-Antwerp, Belgium

Dag Hanstorp

Göteborgs University, Department of Physics, SE-412 96 Göteborg, Sweden

J. Biomed. Opt. 12(4), 044009 (July 24, 2007). doi:10.1117/1.2753478
History: Received July 24, 2006; Revised April 18, 2007; Accepted April 18, 2007; Published July 24, 2007
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We describe the possibility of using a microresonance Raman spectrometer combined with a microfluidic system and optical tweezers to study Escherichia coli (E. coli) overexpressing wild type (wt) neuroglobin (NGB) and its E7Leu mutant, respectively. NGB is a recently discovered heme protein and its function still is a matter of debate. So far, the protein has been studied in its purified form, and in vivo measurements on the single cell level could give more information. To study the feasibility of the combined techniques, the possibilities of the setup are investigated by taking spectra from single cells and clusters of cells. We find that the microresonance Raman technique enables studies of the wt NGB protein in a living cell under fluctuating aerobic and anaerobic conditions. E. coli cells overexpressing wt NGB are stable, and the reversible oxygenation-deoxygenation can be studied over a long period of time. Further, the experiment indicates the presence of an enzymatic system in the bacteria reducing the ferric form NGB. The study of E. coli cells overexpressing E7Leu NGB, on the other hand, gives insight into limiting factors of the setup, such as cell lysis, photoinduced chemistry, and protein concentrations.

Figures in this Article
© 2007 Society of Photo-Optical Instrumentation Engineers

Citation

Kerstin Ramser ; Wim Wenseleers ; Sylvia Dewilde ; Sabine Van Doorslaer ; Luc Moens, et al.
"Micro-resonance Raman study of optically trapped Escherichia coli cells overexpressing human neuroglobin", J. Biomed. Opt. 12(4), 044009 (July 24, 2007). ; http://dx.doi.org/10.1117/1.2753478


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