Digital Microfluidics for Single Bacteria Capture and Selective Retrieval Using Optical Tweezers

When screening microbial populations or consortia for interesting cells, their selective retrieval for further study can be of great interest. To this end, traditional fluorescence activated cell sorting (FACS) and optical tweezers (OT) enabled methods have typically been used. However, the former, although allowing cell sorting, fails to track dynamic cell behavior, while the latter has been limited to complex channel-based microfluidic platforms. In this study, digital microfluidics (DMF) was integrated with OT for selective trapping, relocation, and further proliferation of single bacterial cells, while offering continuous imaging of cells to evaluate dynamic cell behavior. To enable this, magnetic beads coated with Salmonella Typhimurium-targeting antibodies were seeded in the microwell array of the DMF platform, and used to capture single cells of a fluorescent S. Typhimurium population. Next, OT were used to select a bead with a bacterium of interest, based on its fluorescent expression, and to relocate this bead to a different microwell on the same or different array. Using an agar patch affixed on top, the relocated bacterium was subsequently allowed to proliferate. Our OT-integrated DMF platform thus successfully enabled selective trapping, retrieval, relocation, and proliferation of bacteria of interest at single-cell level, thereby enabling their downstream analysis.

[1]  Patricia Yang Liu,et al.  Massive nanophotonic trapping and alignment of rod-shaped bacteria for parallel single-cell studies , 2020 .

[2]  A. Tripathi,et al.  Centrifugal Microfluidics Traps for Parallel Isolation and Imaging of Single Cells , 2020, Micromachines.

[3]  M. Saito,et al.  Real-Time Monitoring and Detection of Single-Cell Level Cytokine Secretion Using LSPR Technology , 2020, Micromachines.

[4]  Anna Bezryadina,et al.  Manipulation and Assessment of Human Red Blood Cells with Tunable “Tug-of-War” Optical Tweezers , 2019 .

[5]  Igor Meglinski,et al.  Influence of Pulsed He–Ne Laser Irradiation on the Red Blood Cell Interaction Studied by Optical Tweezers , 2019, Micromachines.

[6]  Matthias Heinemann,et al.  Manipulating rod-shaped bacteria with optical tweezers , 2019, Scientific Reports.

[7]  Sander K. Govers,et al.  Bimodal Expression of the Salmonella Typhimurium spv Operon , 2018, Genetics.

[8]  Sander K. Govers,et al.  Protein aggregates encode epigenetic memory of stressful encounters in individual Escherichia coli cells , 2018, PLoS biology.

[9]  Lynn Paterson,et al.  Single Cell Isolation Using Optical Tweezers , 2018, Micromachines.

[10]  S. Weissman,et al.  Evolution and heterogeneity of non-hereditary colorectal cancer revealed by single-cell exome sequencing , 2017, Oncogene.

[11]  N. Allbritton,et al.  Automated microraft platform to identify and collect non-adherent cells successfully gene-edited with CRISPR-Cas9. , 2017, Biosensors & bioelectronics.

[12]  David B. Camarillo,et al.  Microfluidic analysis of oocyte and embryo biomechanical properties to improve outcomes in assisted reproductive technologies , 2017, Molecular human reproduction.

[13]  N. Beerenwinkel,et al.  Advances in understanding tumour evolution through single-cell sequencing* , 2017, Biochimica et biophysica acta. Reviews on cancer.

[14]  F. Keplinger,et al.  Microfluidic platform with integrated GMR sensors for quantification of cancer cells , 2017 .

[15]  Dong Sun,et al.  Characterization of Drug Effect on Leukemia Cells Through Single Cell Assay With Optical Tweezers and Dielectrophoresis , 2016, IEEE Transactions on NanoBioscience.

[16]  Stephen R Quake,et al.  Single-cell multimodal profiling reveals cellular epigenetic heterogeneity , 2016, Nature Methods.

[17]  P. Goos,et al.  Optical Manipulation of Single Magnetic Beads in a Microwell Array on a Digital Microfluidic Chip. , 2016, Analytical chemistry.

[18]  Kimberly M. Davis,et al.  Defining heterogeneity within bacterial populations via single cell approaches , 2016, BioEssays : news and reviews in molecular, cellular and developmental biology.

[19]  D. Holden,et al.  Elucidating population-wide mycobacterial replication dynamics at the single-cell level , 2016, Microbiology.

[20]  W. Reik,et al.  Single-cell epigenomics: powerful new methods for understanding gene regulation and cell identity , 2016, Genome Biology.

[21]  Aaron R. Wheeler,et al.  Digital microfluidic immunocytochemistry in single cells , 2015, Nature Communications.

[22]  J. Lammertyn,et al.  Digital microfluidics for time-resolved cytotoxicity studies on single non-adherent yeast cells. , 2015, Lab on a chip.

[23]  Jeroen Lammertyn,et al.  MAGNETIC PARTICLE RETRIEVAL AND POSITIONING IN A MICROWELL ARRAY BY INTEGRATING OPTICAL TWEEZERS IN A DIGITAL MICROFLUIDIC PLATFORM , 2014 .

[24]  I. Nemenman,et al.  Cellular noise and information transmission. , 2014, Current opinion in biotechnology.

[25]  S. Esmaeili,et al.  THE EFFECT OF SOME COSOLVENTS AND SURFACTANTS ON VIABILITY OF CANCEROUS CELL LINES , 2014 .

[26]  Shawn M. Gillespie,et al.  Single-cell RNA-seq highlights intratumoral heterogeneity in primary glioblastoma , 2014, Science.

[27]  A. Aertsen,et al.  Isolation and Validation of an Endogenous Fluorescent Nucleoid Reporter in Salmonella Typhimurium , 2014, PloS one.

[28]  Wolfgang Wiechert,et al.  Microfluidic growth chambers with optical tweezers for full spatial single-cell control and analysis of evolving microbes. , 2013, Journal of microbiological methods.

[29]  I. Thompson,et al.  Single‐cell analysis of circulating tumor cells identifies cumulative expression patterns of EMT‐related genes in metastatic prostate cancer , 2013, The Prostate.

[30]  Robert Puers,et al.  Digital microfluidics-enabled single-molecule detection by printing and sealing single magnetic beads in femtoliter droplets. , 2013, Lab on a chip.

[31]  M. Parsek,et al.  Going local: technologies for exploring bacterial microenvironments , 2013, Nature Reviews Microbiology.

[32]  David R. Latulippe,et al.  Microfluidic extraction, stretching and analysis of human chromosomal DNA from single cells. , 2012, Lab on a chip.

[33]  Soo Hyeon Kim,et al.  A single-cell drug efflux assay in bacteria by using a directly accessible femtoliter droplet array. , 2012, Lab on a chip.

[34]  Johan Paulsson,et al.  Random partitioning of molecules at cell division , 2011, Proceedings of the National Academy of Sciences.

[35]  I. Glauche,et al.  Cellular aging leads to functional heterogeneity of hematopoietic stem cells: a modeling perspective , 2011, Aging cell.

[36]  Lloyd M Smith,et al.  Integrated microfluidic device for automated single cell analysis using electrophoretic separation and electrospray ionization mass spectrometry. , 2010, Analytical chemistry.

[37]  Kuo-Kang Liu,et al.  Optical tweezers for single cells , 2008, Journal of The Royal Society Interface.

[38]  K. Neuman,et al.  Single-molecule force spectroscopy: optical tweezers, magnetic tweezers and atomic force microscopy , 2008, Nature Methods.

[39]  A. Aertsen,et al.  Activation of the Salmonella typhimurium Mrr protein. , 2008, Biochemical and biophysical research communications.

[40]  M. Hajmeer,et al.  Impact of sodium chloride on Escherichia coli O157:H7 and Staphylococcus aureus analysed using transmission electron microscopy. , 2006, Food microbiology.

[41]  Michael Eisenstein,et al.  Cell sorting: Divide and conquer , 2006, Nature.

[42]  G. Bertani,et al.  Lysogeny at Mid-Twentieth Century: P1, P2, and Other Experimental Systems , 2004, Journal of bacteriology.

[43]  K. Namba,et al.  Complete atomic model of the bacterial flagellar filament by electron cryomicroscopy , 2003, Nature.

[44]  P. Swain,et al.  Stochastic Gene Expression in a Single Cell , 2002, Science.

[45]  Miles J. Padgett,et al.  Lights, action: Optical tweezers , 2002 .

[46]  D. Hanstorp,et al.  Sorting Out Bacterial Viability with Optical Tweezers , 2000, Journal of bacteriology.

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

[48]  M W Berns,et al.  Use of a laser-induced optical force trap to study chromosome movement on the mitotic spindle. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[49]  R. Buchanan,et al.  Effect of NaCl, pH, temperature, and atmosphere on growth of Salmonella typhimurium in glucose-mineral salts medium , 1987, Applied and environmental microbiology.

[50]  A. Boville,et al.  Surfactants for the Effective Recovery of Salmonella in Fatty Foods. , 1982, Journal of Food Protection.

[51]  J. Lammertyn,et al.  Digital Microfluidics Assisted Sealing of Individual Magnetic Particles in Femtoliter-Sized Reaction Wells for Single-Molecule Detection. , 2017, Methods in molecular biology.

[52]  G. Obermoser,et al.  Flow cytometry. , 2012, The Journal of investigative dermatology.

[53]  M W Berns,et al.  Laser induced cell fusion in combination with optical tweezers: the laser cell fusion trap. , 1991, Cytometry.