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A special issue of Molecules (ISSN 1420-3049). This special issue belongs to the section "Molecular Diversity".

Deadline for manuscript submissions: closed (20 May 2016) | Viewed by 82536

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Prof. Dr. Fan-Gang Tseng

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Guest Editor

Distinguished Professor, Department of Engineering and System Science, National Tsing Hua University (NTHU), Affiliated Research Fellow, Academia Sinica, Hsinchu, Taiwan
Interests: organ on a chip; microfluidic systems; biosensors; CTCs/CTM diagnosis; single cell analysis
Special Issues, Collections and Topics in MDPI journals

Dr. Tuhin Subhra Santra

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Guest Editor

Department of Engineering Design, Indian Institute of Technology Madras, Chennai, India
Interests: MEMS; Bio-NEMS; single- cell technology; biomedical micro/nano devices; micro/nanofluidics; nanomedicine
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Cells play significant roles in our day-to-day life. However, the behaviors of cells or cells with the environment, with their organelles and their intracellular physical/biochemical/biological effects, are still unknown. Micro/Nanofluidic and Bio-MEMS devices are capable to manipulate and detect bio-samples, reagents, or biomolecules at micro/nano scales and well-fulfill the requirements for cellular analysis. These devices are not only useful to simulate physiological environment of cell, cellular transport mechanisms, cell proliferation, differentiation, neural network coordination, and cardiomyocytes synchronization, but it can also easily control the biochemical, physical, and mechanical parameters of cells.

This Special Issue will invite manuscripts conducting research on the integration of micro/nanofluidics and Bio-MEMS devices dealing with cell manipulation, injection, separation, lysis, and cell dynamics, as well as various detection schemes. The role of cellular analysis in system biology, proteomics, genomics, metabolomics, and their application in bioprocess engineering, future challenge, advantages, and limitations are also welcome to be included in the manuscripts.

Prof. Dr. Fan-Gang Tseng
Dr. Tuhin Subhra Santra
Guest Editors

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Keywords

microfluidics
nanofluidics
bio-MEMS
nanomedicine
lab-on-a-Chip
life-on-a-Chip
organ on a chip
lab-in-a-Cell
cell chip
micro total analysis (µTAS)
flow cytometry
cell heterogeneity
cell perturbation, interaction, cultivation
cell proteomics, genomics, epigenomes, metabolomics, fluxomics
electroporation, optoporation/photoporation, mechanoporation, sonoporation, microinjection
mechanical characterization
optical characterization
biochemical characterization
system biology

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Published Papers (9 papers)

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Research

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5073 KiB

Open AccessArticle

Modulation of Neural Network Activity through Single Cell Ablation: An in Vitro Model of Minimally Invasive Neurosurgery

by Alessandro Soloperto, Marta Bisio, Gemma Palazzolo, Michela Chiappalone, Paolo Bonifazi and Francesco Difato

Molecules 2016, 21(8), 1018; https://doi.org/10.3390/molecules21081018 - 5 Aug 2016

Cited by 6 | Viewed by 5262

Abstract

The technological advancement of optical approaches, and the growth of their applications in neuroscience, has allowed investigations of the physio-pathology of neural networks at a single cell level. Therefore, better understanding the role of single neurons in the onset and progression of neurodegenerative conditions has resulted in a strong demand for surgical tools operating with single cell resolution. Optical systems already provide subcellular resolution to monitor and manipulate living tissues, and thus allow understanding the potentiality of surgery actuated at single cell level. In the present work, we report an in vitro experimental model of minimally invasive surgery applied on neuronal cultures expressing a genetically encoded calcium sensor. The experimental protocol entails the continuous monitoring of the network activity before and after the ablation of a single neuron, to provide a robust evaluation of the induced changes in the network activity. We report that in subpopulations of about 1000 neurons, even the ablation of a single unit produces a reduction of the overall network activity. The reported protocol represents a simple and cost effective model to study the efficacy of single-cell surgery, and it could represent a test-bed to study surgical procedures circumventing the abrupt and complete tissue removal in pathological conditions. Full article

(This article belongs to the Special Issue Micro/Nano Fluidics and Bio-MEMS)

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2088 KiB

Open AccessArticle

Influences of Adhesion Variability on the “Living” Dynamics of Filamentous Bacteria in Microfluidic Channels

by Justin P. Jahnke, Jessica L. Terrell, Austin M. Smith, Xuanhong Cheng and Dimitra N. Stratis-Cullum

Molecules 2016, 21(8), 985; https://doi.org/10.3390/molecules21080985 - 28 Jul 2016

Cited by 9 | Viewed by 8205

Abstract

Microfabricated devices have increasingly incorporated bacterial cells for microscale studies and exploiting cell-based functions in situ. However, the role of surface interactions in controlling the bacterial cell behavior is not well understood. In this study, microfluidic substrates of varied bacterial-binding affinity were used to probe the interaction-driven behavior of filamentous Escherichia coli. In particular, cell alignment under controlled shear flow as well as subsequent orientation and filamentation were compared between cells presenting distinct outer membrane phenotypes. We demonstrated that filaments retained position under flow, which allowed for dynamic single-cell monitoring with in situ elongation of over 100 μm for adherent cells. This maximum was not reached by planktonic cells and was, therefore, adhesion-dependent. The bound filaments initially aligned with flow under a range of flow rates and their continual elongation was traced in terms of length and growth path; analysis demonstrated that fimbriae-mediated adhesion increased growth rate, increased terminal length, as well as dramatically changed the adherent geometry, particularly buckling behavior. The effects to filament length and buckling were further exaggerated by the strongest, specificity-driven adhesion tested. Such surface-guided control of the elongation process may be valuable to yield interesting “living” filamentous structures in microdevices. In addition, this work may offer a biomedically relevant platform for further elucidation of filamentation as an immune-resistant morphology. Overall, this work should inspire broader exploration of microfabricated devices for the study and application of single bacterial cells. Full article

(This article belongs to the Special Issue Micro/Nano Fluidics and Bio-MEMS)

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2142 KiB

Open AccessArticle

A High-Throughput Automated Microfluidic Platform for Calcium Imaging of Taste Sensing

by Yi-Hsing Hsiao, Chia-Hsien Hsu and Chihchen Chen

Molecules 2016, 21(7), 896; https://doi.org/10.3390/molecules21070896 - 8 Jul 2016

Cited by 6 | Viewed by 6953

Abstract

The human enteroendocrine L cell line NCI-H716, expressing taste receptors and taste signaling elements, constitutes a unique model for the studies of cellular responses to glucose, appetite regulation, gastrointestinal motility, and insulin secretion. Targeting these gut taste receptors may provide novel treatments for diabetes and obesity. However, NCI-H716 cells are cultured in suspension and tend to form multicellular aggregates, preventing high-throughput calcium imaging due to interferences caused by laborious immobilization and stimulus delivery procedures. Here, we have developed an automated microfluidic platform that is capable of trapping more than 500 single cells into microwells with a loading efficiency of 77% within two minutes, delivering multiple chemical stimuli and performing calcium imaging with enhanced spatial and temporal resolutions when compared to bath perfusion systems. Results revealed the presence of heterogeneity in cellular responses to the type, concentration, and order of applied sweet and bitter stimuli. Sucralose and denatonium benzoate elicited robust increases in the intracellular Ca²⁺ concentration. However, glucose evoked a rapid elevation of intracellular Ca²⁺ followed by reduced responses to subsequent glucose stimulation. Using Gymnema sylvestre as a blocking agent for the sweet taste receptor confirmed that different taste receptors were utilized for sweet and bitter tastes. This automated microfluidic platform is cost-effective, easy to fabricate and operate, and may be generally applicable for high-throughput and high-content single-cell analysis and drug screening. Full article

(This article belongs to the Special Issue Micro/Nano Fluidics and Bio-MEMS)

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16604 KiB

Open AccessArticle

A PDMS-Based Microfluidic Hanging Drop Chip for Embryoid Body Formation

by Huei-Wen Wu, Yi-Hsing Hsiao, Chih-Chen Chen, Shaw-Fang Yet and Chia-Hsien Hsu

Molecules 2016, 21(7), 882; https://doi.org/10.3390/molecules21070882 - 6 Jul 2016

Cited by 30 | Viewed by 9171

Abstract

The conventional hanging drop technique is the most widely used method for embryoid body (EB) formation. However, this method is labor intensive and limited by the difficulty in exchanging the medium. Here, we report a microfluidic chip-based approach for high-throughput formation of EBs. The device consists of microfluidic channels with 6 × 12 opening wells in PDMS supported by a glass substrate. The PDMS channels were fabricated by replicating polydimethyl-siloxane (PDMS) from SU-8 mold. The droplet formation in the chip was tested with different hydrostatic pressures to obtain optimal operation pressures for the wells with 1000 μm diameter openings. The droplets formed at the opening wells were used to culture mouse embryonic stem cells which could subsequently developed into EBs in the hanging droplets. This device also allows for medium exchange of the hanging droplets making it possible to perform immunochemistry staining and characterize EBs on chip. Full article

(This article belongs to the Special Issue Micro/Nano Fluidics and Bio-MEMS)

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2959 KiB

Open AccessArticle

The Deformation of Polydimethylsiloxane (PDMS) Microfluidic Channels Filled with Embedded Circular Obstacles under Certain Circumstances

by Changhyun Roh, Jaewoong Lee and Chankyu Kang

Molecules 2016, 21(6), 798; https://doi.org/10.3390/molecules21060798 - 18 Jun 2016

Cited by 14 | Viewed by 7933

Abstract

Experimental investigations were conducted to determine the influence of polydimethylsiloxane (PDMS) microfluidic channels containing aligned circular obstacles (with diameters of 172 µm and 132 µm) on the flow velocity and pressure drop under steady-state flow conditions. A significant PDMS bulging was observed when the fluid flow initially contacted the obstacles, but this phenomenon decreased in the 1 mm length of the microfluidic channels when the flow reached a steady-state. This implies that a microfluidic device operating with steady-state flows does not provide fully reliable information, even though less PDMS bulging is observed compared to quasi steady-state flow. Numerical analysis of PDMS bulging using ANSYS Workbench showed a relatively good agreement with the measured data. To verify the influence of PDMS bulging on the pressure drop and flow velocity, theoretical analyses were performed and the results were compared with the experimental results. The measured flow velocity and pressure drop data relatively matched well with the classical prediction under certain circumstances. However, discrepancies were generated and became worse as the microfluidic devices were operated under the following conditions: (1) restricted geometry of the microfluidic channels (i.e., shallow channel height, large diameter of obstacles and a short microchannel length); (2) operation in quasi-steady state flow; (3) increasing flow rates; and (4) decreasing amount of curing agent in the PDMS mixture. Therefore, in order to obtain reliable data a microfluidic device must be operated under appropriate conditions. Full article

(This article belongs to the Special Issue Micro/Nano Fluidics and Bio-MEMS)

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2042 KiB

Open AccessArticle

Comparison of Chip Inlet Geometry in Microfluidic Devices for Cell Studies

by Yung-Shin Sun

Molecules 2016, 21(6), 778; https://doi.org/10.3390/molecules21060778 - 15 Jun 2016

Cited by 18 | Viewed by 5864

Abstract

Micro-fabricated devices integrated with fluidic components provide an in vitro platform for cell studies best mimicking the in vivo micro-environment. These devices are capable of creating precise and controllable surroundings of pH value, temperature, salt concentration, and other physical or chemical stimuli. Various cell studies such as chemotaxis and electrotaxis can be performed by using such devices. Moreover, microfluidic chips are designed and fabricated for applications in cell separations such as circulating tumor cell (CTC) chips. Usually, there are two most commonly used inlets in connecting the microfluidic chip to sample/reagent loading tubes: the vertical (top-loading) inlet and the parallel (in-line) inlet. Designing this macro-to-micro interface is believed to play an important role in device performance. In this study, by using the commercial COMSOL Multiphysics software, we compared the cell capture behavior in microfluidic devices with different inlet types and sample flow velocities. Three different inlets were constructed: the vertical inlet, the parallel inlet, and the vertically parallel inlet. We investigated the velocity field, the flow streamline, the cell capture rate, and the laminar shear stress in these inlets. It was concluded that the inlet should be designed depending on the experimental purpose, i.e., one wants to maximize or minimize cell capture. Also, although increasing the flow velocity could reduce cell sedimentation, too high shear stresses are thought harmful to cells. Our findings indicate that the inlet design and flow velocity are crucial and should be well considered in fabricating microfluidic devices for cell studies. Full article

(This article belongs to the Special Issue Micro/Nano Fluidics and Bio-MEMS)

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Review

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4296 KiB

Open AccessReview

Cardiac Meets Skeletal: What’s New in Microfluidic Models for Muscle Tissue Engineering

by Roberta Visone, Mara Gilardi, Anna Marsano, Marco Rasponi, Simone Bersini and Matteo Moretti

Molecules 2016, 21(9), 1128; https://doi.org/10.3390/molecules21091128 - 26 Aug 2016

Cited by 42 | Viewed by 14450

Abstract

In the last few years microfluidics and microfabrication technique principles have been extensively exploited for biomedical applications. In this framework, organs-on-a-chip represent promising tools to reproduce key features of functional tissue units within microscale culture chambers. These systems offer the possibility to investigate the effects of biochemical, mechanical, and electrical stimulations, which are usually applied to enhance the functionality of the engineered tissues. Since the functionality of muscle tissues relies on the 3D organization and on the perfect coupling between electrochemical stimulation and mechanical contraction, great efforts have been devoted to generate biomimetic skeletal and cardiac systems to allow high-throughput pathophysiological studies and drug screening. This review critically analyzes microfluidic platforms that were designed for skeletal and cardiac muscle tissue engineering. Our aim is to highlight which specific features of the engineered systems promoted a typical reorganization of the engineered construct and to discuss how promising design solutions exploited for skeletal muscle models could be applied to improve cardiac tissue models and vice versa. Full article

(This article belongs to the Special Issue Micro/Nano Fluidics and Bio-MEMS)

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4272 KiB

Open AccessReview

Microfluidic Devices in Advanced Caenorhabditis elegans Research

by Muniesh Muthaiyan Shanmugam and Tuhin Subhra Santra

Molecules 2016, 21(8), 1006; https://doi.org/10.3390/molecules21081006 - 2 Aug 2016

Cited by 26 | Viewed by 8429

Abstract

The study of model organisms is very important in view of their potential for application to human therapeutic uses. One such model organism is the nematode worm, Caenorhabditis elegans. As a nematode, C. elegans have ~65% similarity with human disease genes and, therefore, studies on C. elegans can be translated to human, as well as, C. elegans can be used in the study of different types of parasitic worms that infect other living organisms. In the past decade, many efforts have been undertaken to establish interdisciplinary research collaborations between biologists, physicists and engineers in order to develop microfluidic devices to study the biology of C. elegans. Microfluidic devices with the power to manipulate and detect bio-samples, regents or biomolecules in micro-scale environments can well fulfill the requirement to handle worms under proper laboratory conditions, thereby significantly increasing research productivity and knowledge. The recent development of different kinds of microfluidic devices with ultra-high throughput platforms has enabled researchers to carry out worm population studies. Microfluidic devices primarily comprises of chambers, channels and valves, wherein worms can be cultured, immobilized, imaged, etc. Microfluidic devices have been adapted to study various worm behaviors, including that deepen our understanding of neuromuscular connectivity and functions. This review will provide a clear account of the vital involvement of microfluidic devices in worm biology. Full article

(This article belongs to the Special Issue Micro/Nano Fluidics and Bio-MEMS)

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2480 KiB

Open AccessReview

Development of Droplet Microfluidics Enabling High-Throughput Single-Cell Analysis

by Na Wen, Zhan Zhao, Beiyuan Fan, Deyong Chen, Dong Men, Junbo Wang and Jian Chen

Molecules 2016, 21(7), 881; https://doi.org/10.3390/molecules21070881 - 5 Jul 2016

Cited by 82 | Viewed by 15289

Abstract

This article reviews recent developments in droplet microfluidics enabling high-throughput single-cell analysis. Five key aspects in this field are included in this review: (1) prototype demonstration of single-cell encapsulation in microfluidic droplets; (2) technical improvements of single-cell encapsulation in microfluidic droplets; (3) microfluidic droplets enabling single-cell proteomic analysis; (4) microfluidic droplets enabling single-cell genomic analysis; and (5) integrated microfluidic droplet systems enabling single-cell screening. We examine the advantages and limitations of each technique and discuss future research opportunities by focusing on key performances of throughput, multifunctionality, and absolute quantification. Full article

(This article belongs to the Special Issue Micro/Nano Fluidics and Bio-MEMS)

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