An integrated microfluidic device for the sorting of yeast cells using image processing

The process of detection and separation of yeast cells based on their morphological characteristics is critical to the understanding of cell division cycles, which is of vital importance to the understanding of some diseases such as cancer. The traditional process of manual detection is usually tedious and inconsistent. This paper presents a microfluidic device integrated with microvalves for fluid control for the sorting of yeast cells using image processing algorithms and confirmation based on their fluorescent tag. The proposed device is completely automated, low cost and easy to implement in an academic research setting. Design details of the integrated microfluidic system are highlighted in this paper, along with experimental validation. Real time cell sorting was demonstrated with a cell detection rate of 12 cells per minute.

[1]  Paul J. Choi,et al.  Quantifying E. coli Proteome and Transcriptome with Single-Molecule Sensitivity in Single Cells , 2010, Science.

[2]  Che-Hsin Lin,et al.  Micromachine-based multi-channel flow cytometers for cell/particle counting and sorting , 2005 .

[3]  S. Quake,et al.  A microfabricated fluorescence-activated cell sorter , 1999, Nature Biotechnology.

[4]  Gwo-Bin Lee,et al.  Optically induced flow cytometry for continuous microparticle counting and sorting. , 2008, Biosensors & bioelectronics.

[5]  Bo Yang Yu,et al.  Image processing and classification algorithm for yeast cell morphology in a microfluidic chip. , 2011, Journal of biomedical optics.

[6]  Jessica Melin,et al.  Microfluidic large-scale integration: the evolution of design rules for biological automation. , 2007, Annual review of biophysics and biomolecular structure.

[7]  Dong Sun,et al.  Enhanced cell sorting and manipulation with combined optical tweezer and microfluidic chip technologies. , 2011, Lab on a chip.

[8]  C. Hansen,et al.  Microfluidic single cell analysis: from promise to practice. , 2012, Current opinion in chemical biology.

[9]  K. Mogensen,et al.  Measurements of scattered light on a microchip flow cytometer with integrated polymer based optical elements. , 2004, Lab on a chip.

[10]  B G Schutt The promise in practice. , 1967, The American journal of nursing.

[11]  Robert C. Wolpert,et al.  A Review of the , 1985 .

[12]  S. Quake,et al.  An Integrated Microfabricated Cell Sorter , 2022 .

[13]  Bo Huang,et al.  Counting Low-Copy Number Proteins in a Single Cell , 2007, Science.

[14]  Samuel K Sia,et al.  Lab-on-a-chip devices for global health: past studies and future opportunities. , 2007, Lab on a chip.

[15]  C. Chin,et al.  Lab-ona-chip devices for global health : Past studies and future opportunities , 2006 .

[16]  Suresh V. Garimella,et al.  Recent advances in microscale pumping technologies: a review and evaluation , 2008 .

[17]  Caglar Elbuken,et al.  Microfluidic system with integrated electroosmotic pumps, concentration gradient generator and fish cell line (RTgill-W1)--towards water toxicity testing. , 2009, Lab on a chip.

[18]  Josef Kittler,et al.  Pattern recognition : a statistical approach , 1982 .

[19]  Katherine C. Chen,et al.  Integrative analysis of cell cycle control in budding yeast. , 2004, Molecular biology of the cell.

[20]  J. Diffley,et al.  The Cdc7 protein kinase is required for origin firing during S phase. , 1998, Genes & development.

[21]  Matthias Heinemann,et al.  Whole lifespan microscopic observation of budding yeast aging through a microfluidic dissection platform , 2012, Proceedings of the National Academy of Sciences.

[22]  Juan G. Santiago,et al.  A review of micropumps , 2004 .

[23]  I. Shmulevich,et al.  High-throughput tracking of single yeast cells in a microfluidic imaging matrix. , 2011, Lab on a chip.

[24]  Jeffrey S Erickson,et al.  The good, the bad, and the tiny: a review of microflow cytometry , 2008, Analytical and bioanalytical chemistry.

[25]  Yuejun Kang,et al.  On-chip fluorescence-activated particle counting and sorting system. , 2008, Analytica chimica acta.

[26]  Fabian Rudolf,et al.  Characterization of Single Yeast Cell Phenotypes Using Microfluidic Impedance Cytometry and Optical Imaging , 2016 .

[27]  Christoph A. Merten,et al.  Drop-based microfluidic devices for encapsulation of single cells. , 2008, Lab on a chip.

[28]  S. Quake,et al.  Monolithic microfabricated valves and pumps by multilayer soft lithography. , 2000, Science.

[29]  Anders S Hansen,et al.  High-throughput microfluidics to control and measure signaling dynamics in single yeast cells , 2015, Nature Protocols.

[30]  Gwo-Bin Lee,et al.  Electrokinetically driven micro flow cytometers with integrated fiber optics for on-line cell/particle detection , 2004 .

[31]  Hansen Bow,et al.  Microfluidics for cell separation , 2010, Medical & Biological Engineering & Computing.

[32]  S. Gasser,et al.  A role for the Cdc7 kinase regulatory subunit Dbf4p in the formation of initiation-competent origins of replication. , 1999, Genes & development.

[33]  Hywel Morgan,et al.  Single-cell microfluidic impedance cytometry: a review , 2010 .

[34]  B.P. Ingalls,et al.  Systems Level Modeling of the Cell Cycle Using Budding Yeast , 2007, Cancer informatics.

[35]  V. Lien,et al.  Integrated Fluidic Photonics for Multi-Parameter In-Plane Detection in Microfluidic Flow Cytometry , 2006, LEOS 2006 - 19th Annual Meeting of the IEEE Lasers and Electro-Optics Society.

[36]  Shinsuke Ohnuki,et al.  A microfluidic device to acquire high-magnification microphotographs of yeast cells , 2009, Cell Division.

[37]  Bert R. Meijboom,et al.  Review and Evaluation , 1987 .

[38]  J. Voldman,et al.  Intuitive, image-based cell sorting using optofluidic cell sorting. , 2007, Analytical chemistry.

[39]  Anindya Dutta,et al.  DNA replication in eukaryotic cells. , 2002, Annual review of biochemistry.