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Capillary-assisted Microfluidic Biosensing Platform Captures Single Cell Secretion Dynamics In Nanoliter Compartments.

Amin Hassanzadeh-Barforoushi, Majid Ebrahimi Warkiani, David Gallego-Ortega, Guozhen Liu, Tracie Barber
Published 2020 · Chemistry, Medicine
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Cancer cells continuously secrete inflammatory biomolecules which play significant roles in disease progression and tumor metastasis toward secondary sites. Despite recent efforts to capture cancer cells' intercellular secretion heterogeneity using microfluidics, the challenges in operation of these systems as well as the complexity of designing a biosensing assay for long-term and real-time measurement of single cell secretions have become grand research barriers. Here, we present a new capillary-based microfluidic biosensing approach to easily and reliably capture ~500 single cells inside isolated dead-end nanoliter compartments using simple pipette injection, and quantify their individual secretion dynamics at the single cell resolution over a long period of culture (~16 h). We first present a detailed investigation of the fluid mechanics underlying the formation of nanoliter compartments in the microfluidic system. Based on the measurement of single cell capture efficiency, we employ a one-step FRET-based biosensor which monitors the single cancer cells' protease activity. The sensor reports the fluorescent signal as a product of amino acid chain cleavage and reduction in its quenching capability. Using the single cell protease secretion data, we identified modes of cell secretion dynamics in our cell sample. While most of the cells had low secretion levels, two other smaller and more aggressive secretion dynamics were cells with secretion modes that include sharp spikes or slow but progressive trend. The method presented here overcomes the difficulties associated with performing single cell secretion assays, enabling a feasible and reliable technique for high throughput measurement of metabolic activities in cancer cells.
This paper references
10.1038/nmeth.4154
Sequencing thousands of single-cell genomes with combinatorial indexing
Sarah A. Vitak (2017)
10.1016/j.bios.2019.01.005
Dynamic fluorescent imaging analysis of mitochondrial redox in single cells with a microfluidic device.
Qingling Li (2019)
10.1016/j.copbio.2011.11.023
Single-cell protein analysis.
Meiye Wu (2012)
10.1021/acs.analchem.6b04039
Droplet Microfluidic System with On-Demand Trapping and Releasing of Droplet for Drug Screening Applications.
Matthew Gordon Ray Courtney (2017)
10.1371/journal.pone.0133769
Measurement of Separase Proteolytic Activity in Single Living Cells by a Fluorogenic Flow Cytometry Assay
Wiltrud Haass (2015)
10.1016/j.isci.2019.09.019
A Nanoparticle-Based Affinity Sensor that Identifies and Selects Highly Cytokine-Secreting Cells
Guozhen Liu (2019)
10.1016/j.bbrc.2017.04.097
Detection of cAMP and of PKA activity in Saccharomyces cerevisiae single cells using Fluorescence Resonance Energy Transfer (FRET) probes.
Sonia Colombo (2017)
10.1016/j.celrep.2015.11.048
Quantification of Protein Levels in Single Living Cells.
Chiu-An Lo (2015)
10.1073/pnas.1404472111
Stationary nanoliter droplet array with a substrate of choice for single adherent/nonadherent cell incubation and analysis
Jonathan Shemesh (2014)
10.1007/s10571-018-0620-7
A Novel Microfluidic Device-Based Neurite Outgrowth Inhibition Assay Reveals the Neurite Outgrowth-Promoting Activity of Tropomyosin Tpm3.1 in Hippocampal Neurons
Holly Stefen (2018)
10.1073/pnas.1803884115
Functional profiling of circulating tumor cells with an integrated vortex capture and single-cell protease activity assay
Manjima Dhar (2018)
10.1039/c8lc00403j
Static droplet array for culturing single live adherent cells in an isolated chemical microenvironment.
Amin Hassanzadeh-Barforoushi (2018)
10.1021/ma802805y
Interfacial Structure and Dynamics of Siloxane Systems: PDMS- Vapor and PDMS- Water
Ahmed E. Ismail (2009)
10.1038/nature24286
DNA sequencing at 40: past, present and future
Jay Shendure (2017)
10.1021/ac1015326
Single-cell MALDI-MS as an analytical tool for studying intrapopulation metabolic heterogeneity of unicellular organisms.
Andrea Amantonico (2010)
10.1039/C9ME00047J
IFN-γ-induced signal-on fluorescence aptasensors: from hybridization chain reaction amplification to 3D optical fiber sensing interface towards a deployable device for cytokine sensing
Fuyuan Zhang (2019)
10.1038/nature09145
Single-cell NF-κB dynamics reveal digital activation and analogue information processing
Savaş Tay (2010)
10.1038/srep39846
Isolation of cell type-specific apoptotic bodies by fluorescence-activated cell sorting
Georgia K. Atkin-Smith (2017)
10.1038/nbt.3697
Drug sensitivity of single cancer cells is predicted by changes in mass accumulation rate
Mark M. Stevens (2016)
10.1186/1471-2407-9-188
Expression of matrix metalloproteinases (MMPs) in primary human breast cancer and breast cancer cell lines: New findings and review of the literature
Andrea Köhrmann (2008)
10.1007/S10404-014-1343-9
Unsteady pulsating characteristics of the fluid flow through a sudden expansion microvalve
Amir Nejat (2014)
10.1038/srep11937
A High Throughput Micro-Chamber Array Device for Single Cell Clonal Cultivation and Tumor Heterogeneity Analysis
Feng-Min Shen (2015)
10.3389/fimmu.2019.00755
Techniques for Studying Decoding of Single Cell Dynamics
Stevan Jeknić (2019)
10.1038/srep04736
Real-time single-cell imaging of protein secretion
Yoshitaka Shirasaki (2014)
10.1006/abio.1993.1291
A high throughput fluorogenic substrate for interstitial collagenase (MMP-1) and gelatinase (MMP-9).
David M Bickett (1993)
10.1038/srep35618
A rapid co-culture stamping device for studying intercellular communication
Amin Hassanzadeh-Barforoushi (2016)
10.1039/b802941e
Drop-based microfluidic devices for encapsulation of single cells.
Sarah Koester (2008)
10.1242/dev.101063
Dynamic stem cell heterogeneity
Teresa G. Krieger (2015)
10.1002/pmic.201600370
Proteomic Profiling of Exosomes Secreted by Breast Cancer Cells with Varying Metastatic Potential.
Lahiru Gangoda (2017)
10.1039/c7lc00846e
Staggered trap arrays for robust microfluidic sample digitization.
Alejandra Sposito (2017)
10.1016/J.TRAC.2018.03.002
Advances in structure-switching aptasensing towards real time detection of cytokines
Chaomin Cao (2018)
10.1039/c3lc50953b
On-chip regeneration of aptasensors for monitoring cell secretion.
Qing Zhou (2014)
10.1038/nature08872
Understanding mechanisms underlying human gene expression variation with RNA sequencing
Joseph K. Pickrell (2010)
10.1016/j.bios.2014.11.001
Jetting microfluidics with size-sorting capability for single-cell protease detection.
Tengyang Jing (2015)
10.1016/j.cell.2015.05.002
Highly Parallel Genome-wide Expression Profiling of Individual Cells Using Nanoliter Droplets
Evan Z. Macosko (2015)
10.1063/1.4974904
An easily fabricated three-dimensional threaded lemniscate-shaped micromixer for a wide range of flow rates.
Mehdi Rafeie (2017)
10.1002/advs.201801361
Multiplexed, Sequential Secretion Analysis of the Same Single Cells Reveals Distinct Effector Response Dynamics Dependent on the Initial Basal State
Zhuo Chen (2019)
10.1016/j.bios.2010.09.006
Droplet-based microfluidic platforms for single T cell secretion analysis of IL-10 cytokine.
Tania Konry (2011)
10.1016/j.gendis.2014.12.002
Targeting matrix metalloproteinases in cancer: Bringing new life to old ideas
Jillian Cathcart (2015)
10.1039/c3lc50945a
Probing cellular heterogeneity in cytokine-secreting immune cells using droplet-based microfluidics.
Venkatachalam Chokkalingam (2013)
10.1039/c0ib00083c
Proteolytic Activity Matrix Analysis (PrAMA) for simultaneous determination of multiple protease activities.
Miles A. Miller (2011)
10.1039/c5lc00614g
The Poisson distribution and beyond: methods for microfluidic droplet production and single cell encapsulation.
David J. Collins (2015)
10.1073/pnas.1814348116
Multiplexed profiling of single-cell extracellular vesicles secretion
Yahui Ji (2019)
10.1002/smll.201800698
Label-Free Optofluidic Nanobiosensor Enables Real-Time Analysis of Single-Cell Cytokine Secretion.
Xiaokang Li (2018)
10.1021/acssensors.9b01057
Microfluidic actuation via 3D-printed molds towards multiplex biosensing of cell apoptosis.
Bac Van Dang (2019)
10.1146/annurev-bioeng-090215-112735
Innovative Tools and Technology for Analysis of Single Cells and Cell-Cell Interaction.
Tania Konry (2016)
10.1039/c2lc20876h
A microchamber array for single cell isolation and analysis of intracellular biomolecules.
Klaus Eyer (2012)
10.1016/j.it.2014.03.004
Heterogeneity in immune responses: from populations to single cells.
Rahul Satija (2014)
10.1039/b926849a
Multidimensional analysis of the frequencies and rates of cytokine secretion from single cells by quantitative microengraving.
Qing Han (2010)



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