Single-cell phenotyping is critical to the success of biological reductionism. Raman-activated cell sorting (RACS) has shown promise in resolving the dynamics of living cells at the individual level and to uncover population heterogeneities in comparison to established approaches such as fluorescence-activated cell sorting (FACS). Given that the number of single-cells would be massive in any experiment, the power of Raman profiling technique for single-cell analysis would be fully utilized only when coupled with a high-throughput and intelligent process control and data analysis system. In this work, we established QSpec, an automatic system that supports high-throughput Raman-based single-cell phenotyping. Additionally, a single-cell Raman profile database has been established upon which data-mining could be applied to discover the heterogeneity among single-cells under different conditions. To test the effectiveness of this control and data analysis system, a sub-system was also developed to simulate the phenotypes of single-cells as well as the device features.
References
[1]
Amann RI, Ludwig W, Schleifer KH. 1995. Phylogenetic identification and in-situ detection of individual microbial-cells without cultivation. Microbiological Reviews 59:143-169
[2]
Amantonico A, Urban PL, Fagerer SR, Balabin RM, Zenobi R. 2010. Single-cell MALDI-MS as an analytical tool for studying intrapopulation metabolic heterogeneity of unicellular organisms. Analytical Chemistry 82:7394-7400
[3]
Beja O, Aravind L, Koonin EV, Suzuki MT, Hadd A, Nguyen LP, Jovanovich SB, Gates CM, Feldman RA, Spudich JL, Spudich EN, DeLong EF. 2000. Bacterial rhodopsin: evidence for a new type of phototrophy in the sea. Science 289:1902-1906
Huang WE, Ward AD, Whiteley AS. 2009. Raman tweezers sorting of single microbial cells. Environmental Microbiology Reports 1:44-49
[8]
Lewis IR, Edwards H. 2001. Handbook of Raman spectroscopy: from the research laboratory to the process line, Practical spectroscopy series. New York: Marcel Dekker. Vol. 28
[9]
Li M, Xu J, Romero-Gonzalez M, Banwart SA, Huang WE. 2012. Single cell Raman spectroscopy for cell sorting and imaging. Current Opinion in Biotechnology 23:56-63
[10]
Monson RK, Lipson DL, Burns SP, Turnipseed AA, Delany AC, Williams MW, Schmidt SK. 2006. Winter forest soil respiration controlled by climate and microbial community composition. Nature 439:711-714
[11]
Pascut FC, Goh HT, George V, Denning C, Notingher I. 2011. Toward label-free Raman-activated cell sorting of cardiomyocytes derived from human embryonic stem cells. Journal of Biomedical Optics 16(4):045002
[12]
Rappe MS, Giovannoni SJ. 2003. The uncultured microbial majority. Annual Review of Microbiology 57:369-394
[13]
Snook RD, Harvey TJ, Faria EC, Gardner P. 2009. Raman tweezers and their application to the study of singly trapped eukaryotic cells. Integrative Biology 1:43-52
[14]
Sobel I. 1978. Neighborhood coding of binary images for fast contour following and general binary array processing. Computer Graphics and Image Processing 8:127-135
[15]
Vapnik V. 2000. The nature of statistical learning theory. New York Inc: Springer-Verlag.
[16]
Wagner M. 2009. Single-cell ecophysiology of microbes as revealed by raman microspectroscopy or secondary ion mass spectrometry imaging. Annual Review of Microbiology 63:411-429
[17]
Wang MM, Tu E, Raymond DE, Yang JM, Zhang H, Hagen N, Dees B, Mercer EM, Forster AH, Kariv I, Marchand PJ, Butler WF. 2005. Microfluidic sorting of mammalian cells by optical force switching. Nature Biotechnology 23:83-87
[18]
Whitaker RD, Walt DR. 2007. Multianalyte single-cell analysis with multiple cell lines using a fiber-optic array. Analytical Chemistry 79:9045-9053
[19]
Wlodkowic D, Cooper JM. 2010a. Microfabricated analytical systems for integrated cancer cytomics. Analytical and Bioanalytical Chemistry 398:193-209
[20]
Wlodkowic D, Cooper JM. 2010b. Tumors on chips: oncology meets microfluidics. Current Opinion in Chemical Biology 14:556-567
[21]
Xia W, Zhang C, Zeng X, Feng Y, Weng J, Lin X, Zhu J, Xiong Z, Xu J, Cai Z, Jia Z. 2011. Autotrophic growth of nitrifying community in an agricultural soil. The ISME Journal 5:1226-1236
[22]
Zong C, Lu S, Chapman AR, Xie XS. 2012. Genome-wide detection of single-nucleotide and copy-number variations of a single human cell. Science 338:1622-1626
