A Raman Flow Cytometer: An Innovative Microfluidic Approach for Continuous Label-Free Analysis of Cells via Raman Spectroscopy
AuthorsDe Grazia, Antonio
AdvisorsDi Fabrizio, Enzo M.
KAUST DepartmentPhysical Sciences and Engineering (PSE) Division
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AbstractIn this work a Raman flow cytometer is presented. It is a whole new microfluidic device that takes advantage of basic principles of Raman spectroscopy and fluorescent flow cytometry mixed together in a system of particularly shaped channels. These are indeed composed by specific shape and sizes – thanks to which cells can flow one-by-one – and a trap by means of which cells are trapped in order to perform Raman analysis on single ones in a constant and passive way. In this sense the microfluidic device promotes a fast method to look for single cells in a whole multicellular sample. It is a label-free analysis and this means that, on the contrary of what happens with fluorescent flow cytometry, the sample does not need to undergo any particular time-consuming pretreatment before being analyzed. Moreover it gives a complete information about the biochemical content of the sample thanks to the involvement of Raman spectroscopy as method of analysis. Many thought about a device like this, but eventually it is the first one being designed, fabricated and tested. The materials involved in the production of the Raman flow cytometer are chosen wisely. In particular the chip – the most important component of the device – is multilayered, being composed by a slide of calcium fluoride (which gives a negligible signal in Raman analyses), a photosensitive resist containing a pattern with channels and another slide of calcium fluoride in order for the channels to be sealed on both sides. The chip is, in turn, connected to gaskets and external frames. Several fabrication processes are followed to ultimately get the complete Raman flow cytometer and experiments on red blood cells demonstrate its validity in this field.