Monitoring dynamic protein expression in living E-coli. Bacterial Celts by laser tweezers raman spectroscopy

Chan JW, Winhold H, Corzett MH, Ulloa JM, Cosman M, Balhorn R, Huser T (2007)
Cytometry Part A 71A(7): 468-474.

Journal Article | Published | English

No fulltext has been uploaded

Author
; ; ; ; ; ;
Abstract
Background: Laser tweezers Raman spectroscopy (LTRS) is a novel, nondestructive, and label-free method that can be used to quantitatively measure changes in cellular activity in single living cells. Here, we demonstrate its use to monitor changes in a population of E coli cells that occur during overexpression of a protein, the extracellular domain of myelin oligodendrocyte glycoprotein [MOG(1-120)]. Methods: Raman spectra were acquired from individual E coli cells suspended in solution and trapped by a single tightly focused laser beam. Overexpression of MOG(I-120) in transformed E coli Rosetta-Gami (DE3)pLysS cells was induced by addition of isopropyl thiogalactoside (IPTG). Changes in the peak intensities of the Raman spectra from a population of cells were monitored and analyzed over a total duration of 3 h. Data were also collected for concentrated purified MOG(1-120) protein in solution, and the spectra compared with that obtained for the MOG(I-120) expressing cells. Results: Raman spectra of individual, living E coli cells exhibit signatures due to DNA and protein molecular vibrations. Characteristic Raman markers associated with protein vibrations, such as 1,257, 1,340, 1,453, and 1,660 cm(-1), are shown to increase as a function of time following the addition of IPTG. Comparison of these spectra and the spectra of purified MOG protein indicates that the changes are predominantly due to the induction of MOG protein expression. Protein expression was found to occur mostly within the second hour, with a 470% increase relative to the protein expressed in the first hour. A 230% relative increase between the second and third hour indicates that protein expression begins to level off within the third hour. Conclusion: It is demonstrated that LTRS has sufficient sensitivity for real-time, nondestructive, and quantitative monitoring of biological processes, such as protein expression, in single living cells. Such capabilities, which are not: currently available in flow cytometry, open up new possibilities for analyzing cellular processes occurring in single microbial and eukaryotic cells, Published 2007 Wiley-Liss, Inc.
Publishing Year
ISSN
eISSN
PUB-ID

Cite this

Chan JW, Winhold H, Corzett MH, et al. Monitoring dynamic protein expression in living E-coli. Bacterial Celts by laser tweezers raman spectroscopy. Cytometry Part A. 2007;71A(7):468-474.
Chan, J. W., Winhold, H., Corzett, M. H., Ulloa, J. M., Cosman, M., Balhorn, R., & Huser, T. (2007). Monitoring dynamic protein expression in living E-coli. Bacterial Celts by laser tweezers raman spectroscopy. Cytometry Part A, 71A(7), 468-474.
Chan, J. W., Winhold, H., Corzett, M. H., Ulloa, J. M., Cosman, M., Balhorn, R., and Huser, T. (2007). Monitoring dynamic protein expression in living E-coli. Bacterial Celts by laser tweezers raman spectroscopy. Cytometry Part A 71A, 468-474.
Chan, J.W., et al., 2007. Monitoring dynamic protein expression in living E-coli. Bacterial Celts by laser tweezers raman spectroscopy. Cytometry Part A, 71A(7), p 468-474.
J.W. Chan, et al., “Monitoring dynamic protein expression in living E-coli. Bacterial Celts by laser tweezers raman spectroscopy”, Cytometry Part A, vol. 71A, 2007, pp. 468-474.
Chan, J.W., Winhold, H., Corzett, M.H., Ulloa, J.M., Cosman, M., Balhorn, R., Huser, T.: Monitoring dynamic protein expression in living E-coli. Bacterial Celts by laser tweezers raman spectroscopy. Cytometry Part A. 71A, 468-474 (2007).
Chan, James W., Winhold, Heiko, Corzett, Michele H., Ulloa, Joshua M., Cosman, Monique, Balhorn, Rod, and Huser, Thomas. “Monitoring dynamic protein expression in living E-coli. Bacterial Celts by laser tweezers raman spectroscopy”. Cytometry Part A 71A.7 (2007): 468-474.
This data publication is cited in the following publications:
This publication cites the following data publications:

10 Citations in Europe PMC

Data provided by Europe PubMed Central.

Raman microspectroscopy detects epigenetic modifications in living Jurkat leukemic cells.
Poplineau M, Trussardi-Regnier A, Happillon T, Dufer J, Manfait M, Bernard P, Piot O, Antonicelli F., Epigenomics 3(6), 2011
PMID: 22126296
Tumour cell identification by means of Raman spectroscopy in combination with optical traps and microfluidic environments.
Dochow S, Krafft C, Neugebauer U, Bocklitz T, Henkel T, Mayer G, Albert J, Popp J., Lab Chip 11(8), 2011
PMID: 21340095
Confocal Raman microscopy of optical-trapped particles in liquids.
Cherney DP, Harris JM., Annu Rev Anal Chem (Palo Alto Calif) 3(), 2010
PMID: 20636043
Raman and CARS microspectroscopy of cells and tissues.
Krafft C, Dietzek B, Popp J., Analyst 134(6), 2009
PMID: 19475129
Single-cell research: what determines its feasibility?
Sabelnikov A, Kempf CR., Anal. Biochem. 383(2), 2008
PMID: 18814839
An integrated optofluidic platform for Raman-activated cell sorting.
Lau AY, Lee LP, Chan JW., Lab Chip 8(7), 2008
PMID: 18584087

16 References

Data provided by Europe PubMed Central.

The human T cell response to myelin oligodendrocyte glycoprotein: a multiple sclerosis family-based study.
Koehler NK, Genain CP, Giesser B, Hauser SL., J. Immunol. 168(11), 2002
PMID: 12023398
Study of dynamical process of heat denaturation in optically trapped single microorganisms by near-infrared Raman spectroscopy
Xie, Journal of Applied Physics 94(9), 2003
Optical-trapping Raman microscopy detection of single unilamellar lipid vesicles.
Cherney DP, Conboy JC, Harris JM., Anal. Chem. 75(23), 2003
PMID: 14640737
Reagentless identification of single bacterial spores in aqueous solution by confocal laser tweezers Raman spectroscopy.
Chan JW, Esposito AP, Talley CE, Hollars CW, Lane SM, Huser T., Anal. Chem. 76(3), 2004
PMID: 14750852
Laser irradiation and Raman spectroscopy of single living cells and chromosomes: sample degradation occurs with 514.5 nm but not with 660 nm laser light.
Puppels GJ, Olminkhof JH, Segers-Nolten GM, Otto C, de Mul FF, Greve J., Exp. Cell Res. 195(2), 1991
PMID: 2070819
Activation-dependent phases of T cells distinguished by use of optical tweezers and near infrared Raman spectroscopy.
Mannie MD, McConnell TJ, Xie C, Li YQ., J. Immunol. Methods 297(1-2), 2005
PMID: 15777930
Identification of single bacterial cells in aqueous solution using confocal laser tweezers Raman spectroscopy.
Xie C, Mace J, Dinno MA, Li YQ, Tang W, Newton RJ, Gemperline PJ., Anal. Chem. 77(14), 2005
PMID: 16013851
Micro-Raman spectroscopy detects individual neoplastic and normal hematopoietic cells.
Chan JW, Taylor DS, Zwerdling T, Lane SM, Ihara K, Huser T., Biophys. J. 90(2), 2006
PMID: 16239327
The lag phase and G1 phase of a single yeast cell monitored by Raman microspectroscopy
Singh, Journal of Raman Spectroscopy 37(8), 2006

Export

0 Marked Publications

Open Data PUB

Web of Science

View record in Web of Science®

Sources

PMID: 17458881
PubMed | Europe PMC

Search this title in

Google Scholar