Abstract
Three important technical innovations are reported here towards Raman-activated cell sorting. Firstly, a microfluidic chip made of quartz is introduced which integrates injection of single cells, trapping by laser fibres and sorting of cells. Secondly, a chip holder was designed to provide simple, accurate and stable adjustment of chips, microfluidic connections and the trapping laser fibres. The new setup enables to the collection of Raman spectra of single cells at 785 nm excitation with 10 s exposure time. Lastly, a new type of modelling the various background contributions is described, improving Raman-based cell identification by the classification algorithm linear discriminant analysis. Mean sensitivity and specificity determined by iterated 10-fold cross validation were 96 and 99 %, respectively, for the distinction of leucocytes extracted from blood, breast cancer cells BT-20 and MCF-7, and leukaemia cells OCI-AML3.
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References
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Dochow S, Krafft C, Neugebauer U, Bocklitz T, Henkel T, Mayer G, Albert J, Popp J (2011) Tumour cell identification by means of Raman spectroscopy in combination with optical traps. Lab Chip 11(8):1484–1490
Dochow S, Beleites C, Henkel T, Mayer G, Albert J, Krafft C, Popp J (2012) Quartz microfluidic chip for tumour cell identification by Raman spectroscopy in combination with optical traps. Lab Chip. doi:10.1039/C2LC41169E, Accepted
Dochow S, Uhlemann W, Krafft C (2012) Justierbare Aufnahmevorrichtung für mikrofluidische Chips mit einzukoppelnder optischer Faser. Germany Patent DE102010050676B3
Acknowledgments
The authors thank C. Jörke (University Hospital Jena) for providing the cells. Financial support from the European Union through the Europäischer Fonds für regionale Entwicklung EFRE and the “Thüringer Ministerium für Bildung, Wissenschaft und Kultur” (project B714-07037) is highly acknowledged.
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Dochow, S., Beleites, C., Henkel, T. et al. Quartz microfluidic chip for tumour cell identification by Raman spectroscopy in combination with optical traps. Anal Bioanal Chem 405, 2743–2746 (2013). https://doi.org/10.1007/s00216-013-6726-3
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DOI: https://doi.org/10.1007/s00216-013-6726-3