QSpec: online control and data analysis system for single-cell Raman spectroscopy.

Lihui Ren,Kang Ning,Jian Xu,Xiaoquan Su,Yun Wang

doi:10.7717/peerj.436

Lihui Ren, Kang Ning + Show 3 more

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https://doi.org/10.7717/peerj.436

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Abstract

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.

Highlights

All organisms on earth, including bacteria, plants and animals, derive from single-cells
We describe our approach for Ramanactivated cell sorting (RACS) system intelligent control and high-throughput data analysis in the following order: (1) Establishment of an automatic high-throughput process control system QSpec that could support the full cycle of single-cell phenotyping: instrument control, single-cell
Materials and assessment methods The RACS system that we used is a prototype that consists of Raman spectroscopy, single-cell sorting, microfluidic device module and QSpec control and data analysis module

Summary

Introduction

All organisms on earth, including bacteria, plants and animals, derive from single-cells. Microbiologists are especially interested in single-cell techniques because most microorganisms (>99%) have not yet been cultured in the lab (Amann, Ludwig & Schleifer, 1995; Rappe & Giovannoni, 2003). These uncultivated microorganisms contain a large amount of functional genes and play crucial roles in natural ecosystems through various ways such as global warming (Monson et al, 2006), food security (through maintaining soil health and promoting plant growth) (Xia et al, 2011), and environmental bioremediation (Beja et al, 2000).

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