Single-cell analysis

Of the mass spectrometric techniques, SIMS has become a useful tool for the imaging of single cells.174 Human liver tumor cells poisoned with Cd, Cu, Cr, Hg and Zn were analyzed using imaging ToF-SIMS (ION-TOF) to visualize the metal distributions on a single cell basis. Among the five metal ions investigated, only Cr and Cu showed a preferential diffusion into the cell after 

To analyze element distribution, isotope ratios and species in single cells, development of advanced inorganic mass spectrometric techniques in combination with biomolecular mass spectrometry is required for future applications. 

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Darvey, H. M. Kell, D. B. Flow cytometry and cell sorting of heterogenous microbial populations the importance of single cell analysis. Microbiol. Rev. 1997, 60, 641-696. 

New Frontiers in Ultrasensitive Bioanalysis Advanced Analytical Chemistry Applications in Nanobiotechnology, Single Molecule Detection, and Single Cell Analysis. 

Capillary electrophoresis (CE) either coupled to MS or to laser-induced fluorescence (LIF) is less often used in metabolomics approaches. This method is faster than the others and needs a smaller sample size, thereby making it especially interesting for single cell analysis [215] The most sensitive mass spectrometers are the Orbitrap and Fourier transform ion cyclotron resonance (FT-ICR) MS [213]. These machines determine the mass-to-charge ratio of a metabolite so accurate that its empirical formula can be predicted, making them the techniques of choice for the identification of unknown peaks. 

Optical fiber detectors (OFD) are devices that measure electromagnetic radiation transmitted through optical fibers to produce a quantitative signal in response to the chemical or biochemical recognition of a specific analyte. Ideally, an OFD should produce a specific and accurate measurement, continuously and reversibly, of the presence of a particular molecular species in a given sample medium. Additionally, OFD should pro vide maximum sensitivity and minimal interferences fromsuperfluous ions or molecules to obtain low detection limits. Other attractive features include the miniaturization of the fiber s tip to accommodate single-cell analysis and portable instrumentation to allow in situ analysis. 

Gavin MA, Torgerson TR, Houston E, DeRoos P, Ho WY, Stray-Pedersen A, Ocheltree EL, Greenberg PD, Ochs HD, Rudensky AY Single-cell analysis of normal and FOXP3-mutant human... 

G. K. Shoemaker, J. Lorieau, L. H. Lau, C. S. Gillmor, and M. M. Palcic, Multiple Sampling in Single-Cell Enzyme Assays, Anal. Chem. 2005, 77, 3132 X. Sun and W. Jin, Catalysis-Electrochemical Determination of Zeptomol Enzyme and Its Application for Single-Cell Analysis, Anal. Chem. 2003, 75, 6050. 

Durack G, Robinson JP, eds. (2000). Emerging Tools for Single Cell Analysis Advances in Optical Measurement Technologies. Wiley-Liss, Inc, New York. 

Sun et al. [59] used NCE-LIF to evaluate a derivatization method mediated by liposome for single cell analysis. According to these workers single cell analysis revealed that liposome-membrane fusion occurred after entrance of liposomes into the cells, with release of encapsulated fluorescence dyes and labeling of intracellular species. 

Andersson, H., van den Berg, A., Microtechnologies and nanotechnologies for single-cell analysis. Curr. Opin. Biotechnol. 2004, 15, 44-49. 

Tamaki, E., Sato, K., Tokeshi, M., Sato, K., Aihara, M., Kitamori, T., Single-cell analysis by a scanning thermal lens microscope with a microchip direct monitoring of cytochrome c distribution during apoptosis process. Anal. Chem. 2002, 74, 1560-1564. 

While a wide range of opportunities exist, such as environmental, clinical, and trace analysis, the principal application for labs-on-a-chip is in the analysis of biological samples. The miniaturized dimensions allow extremely small sample volumes to be analyzed, and a microchip format can allow chemical reaction, mixing, sample manipulation, and multiplexing to be performed. Single-cell analysis, immunoassays, protein and peptide separations, DNA analysis and sequencing, and polymerase chain reactions have all been performed on microchip devices [48]. 

SYSTEMS NANOBIOLOGY FROM QUANTITATIVE SINGLE MOLECULE BIOPHYSICS TO MICROFLUIDIC-BASED SINGLE CELL ANALYSIS... 

In this chapter, three applications how quantitative informations which are representative for cellular processes or single cell analysis like gene expression regulation,... 

Figure 10. Optical setup used for the detection of native UV-fluorescence for microfluidic single cell analysis. The photograph shows a mounted PDMS microstmcture with voltage connections.

Heath, J.R., Phelps, M.E. and Hood, L. (2003) Nanosystems biology. Mol. Imaging Biol. 5, 312-325. Hellmich, W., Gieif, D., Pelaigus, C., Anselmetti, D. and Ros, A. (2006) Improved native laser induced fluorescence detection for single cell analysis in poly(dimethylsiloxane) microfluidic devices. J. Chromatogr. A. 1130, 195-200. 

Systems Nanobiology From Quantitative Single Molecule Biophysics to Microfluidic-Based Single Cell Analysis... 

Cottrell, M. T., and Kirchman, D. L. (2004). Single-cell analysis of bacterial growth, cell size, and community structure in the Delaware River estuary. Aquat. Microb. Ecol. 34, 139—149. 

Voss TC, John S, Hager GL. Single-cell analysis of glucocorticoid receptor action reveals that stochastic post-chromatin association mechanisms regulate Ugand-specific transcription. Mol Endocrinol. 2006 20 2641-2655. 

Zheng F, Qin YJ, Chen K (2007) Sensitivity map of laser tweezers Raman spectroscopy for single-cell analysis of colorectal cancer. J Biomed Opt 12(3) 034002 Chan JW et al (2008) Nondestmctive identification of iudividual leukemia cells by laser trapping Raman spectroscopy. Anal Chem 80(6) 2180-2187... 

Results from studies of biology systems lead to the conclusion that individualized diagnosis, clinical evaluation, and treatment will be inevitable. In many instances, amount of sample available for diagnostic purposes is and will be limited demanding more sensitive techniques and instrumentation on one hand, and development of single cell analysis on the other hand. Automated and simple steps of sample processing combined with highly sensitive and specific measurements will improve what cuiTently contributes to loss of important markers in a limited sample. 

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