Larry J. Millet

University of Tennessee at Knoxville

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Martha U. Gillette

University of Illinois Urbana-Champaign

Rashid Bashir

Illinois College

Gabriel Popescu

University of Illinois Urbana-Champaign

Scott T. Retterer

Oak Ridge National Laboratory

Mitchel J. Doktycz

Oak Ridge National Laboratory

William P. King

University of Illinois Urbana-Champaign

Sandra Davern

Oak Ridge National Laboratory

Zhuo Wang

Beihang University

Jonathan V. Sweedler

University of Illinois Urbana-Champaign

Jayde A. Aufrecht

Environmental Molecular Sciences Laboratory

Cooperative Institutions

University of Illinois Urbana-Champaign

Oak Ridge National Laboratory

University of Tennessee at Knoxville

Urbana University

Oak Ridge Associated Universities

Centre National de la Recherche Scientifique

University of Illinois System

Institute of Micro and Nanotechnology

Purdue University West Lafayette

Inserm

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MULTILAYER PLANT-IN-CHIP PLATFORM FOR NON-DESTRUCTIVE VISUALIZATION OF ROOT MORPHOLOGY DURING CO-CULTURE WITH BENEFICIAL BACTERIA

OSTI OAI (U.S. Department of Energy Office of Scientific and Technical Information) (2017)

Jayde A. Aufrecht Collin M. Timm Jennifer Ryan Amber B. Webb Larry J. Millet

Morphology

Root (linguistics)

Source

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Citations (0)

Measuring Physical Properties of Neuronal and Glial Cells with Resonant Microsensors

Analytical Chemistry (2014)

Elise A. Corbin Larry J. Millet Katrina R. Keller William P. King Rashid Bashir

Microelectromechanical systems (MEMS) resonant sensors provide a high degree of accuracy for measuring the physical properties of chemical and biological samples. These sensors enable the investigation of cellular mass and growth, though previous sensor designs have been limited to the study of homogeneous cell populations. Population heterogeneity, as is generally encountered in primary cultures, reduces measurement yield and limits the efficacy of sensor mass measurements. This paper presents a MEMS resonant pedestal sensor array fabricated over through-wafer pores compatible with vertical flow fields to increase measurement versatility (e.g., fluidic manipulation and throughput) and allow for the measurement of heterogeneous cell populations. Overall, the improved sensor increases capture by 100% at a flow rate of 2 μL/min, as characterized through microbead experiments, while maintaining measurement accuracy. Cell mass measurements of primary mouse hippocampal neurons in vitro, in the range of 0.1-0.9 ng, demonstrate the ability to investigate neuronal mass and changes in mass over time. Using an independent measurement of cell volume, we find cell density to be approximately 1.15 g/mL.

Microbead (research)

Fluidics

10.1021/ac5000625

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Citations (25)

Accessing microfluidics through feature-based design software for 3D printing

PLoS ONE (2018)

Peter G. Shankles Larry J. Millet Jayde A. Aufrecht Scott T. Retterer

Additive manufacturing has been a cornerstone of the product development pipeline for decades, playing an essential role in the creation of both functional and cosmetic prototypes. In recent years, the prospects for distributed and open source manufacturing have grown tremendously. This growth has been enabled by an expanding library of printable materials, low-cost printers, and communities dedicated to platform development. The microfluidics community has embraced this opportunity to integrate 3D printing into the suite of manufacturing strategies used to create novel fluidic architectures. The rapid turnaround time and low cost to implement these strategies in the lab makes 3D printing an attractive alternative to conventional micro- and nanofabrication techniques. In this work, the production of multiple microfluidic architectures using a hybrid 3D printing-soft lithography approach is demonstrated and shown to enable rapid device fabrication with channel dimensions that take advantage of laminar flow characteristics. The fabrication process outlined here is underpinned by the implementation of custom design software with an integrated slicer program that replaces less intuitive computer aided design and slicer software tools. Devices are designed in the program by assembling parameterized microfluidic building blocks. The fabrication process and flow control within 3D printed devices were demonstrated with a gradient generator and two droplet generator designs. Precise control over the printing process allowed 3D microfluidics to be printed in a single step by extruding bridge structures to 'jump-over' channels in the same plane. This strategy was shown to integrate with conventional nanofabrication strategies to simplify the operation of a platform that incorporates both nanoscale features and 3D printed microfluidics.

Rapid Prototyping

10.1371/journal.pone.0192752

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Citations (18)

Separating Beads and Cells in Multi-channel Microfluidic Devices Using Dielectrophoresis and Laminar Flow

Journal of Visualized Experiments (2011)

Larry J. Millet Kidong Park Nicholas N. Watkins K. Jimmy Hsia Rashid Bashir

Microfluidic devices have advanced cell studies by providing a dynamic fluidic environment on the scale of the cell for studying, manipulating, sorting and counting cells. However, manipulating the cell within the fluidic domain remains a challenge and requires complicated fabrication protocols for forming valves and electrodes, or demands specialty equipment like optical tweezers. Here, we demonstrate that conventional printed circuit boards (PCB) can be used for the non-contact manipulation of cells by employing dielectrophoresis (DEP) for bead and cell manipulation in laminar flow fields for bioactuation, and for cell and bead separation in multichannel microfluidic devices. First, we present the protocol for assembling the DEP electrodes and microfluidic devices, and preparing the cells for DEP. Then, we characterize the DEP operation with polystyrene beads. Lastly, we show representative results of bead and cell separation in a multichannel microfluidic device. In summary, DEP is an effective method for manipulating particles (beads or cells) within microfluidic devices.

Dielectrophoresis

Bead

Fluidics

Polystyrene

10.3791/2545-v

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Citations (1)

Hydrogel Microstructures: Characterization of Mass and Swelling of Hydrogel Microstructures using MEMS Resonant Mass Sensor Arrays (Small 16/2012)

Small (2012)

Larry J. Millet Elise A. Corbin Robert Free Kidong Park Hyunjoon Kong

Through photolithography, poly(ethylene glycol)diacrylate microstructures can be attached to microelectromechanical system resonant mass sensor arrays to enable the characterization of hydrogel mass and swelling. Mass values obtained coincide with the values obtained through imaging-based measures of microstructure volume for independent mass determination. The data shown on page 2555 indicate a size-dependent increase in gel mass with increasing fluid density; volume comparisons of a range of bulk hydrogel concentrations (5% to 100% v/v) show a non-linear swelling trend.

Characterization

Mass concentration (chemistry)

10.1002/smll.201290089

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Citations (3)

Guiding neuron development with planar surface gradients of substrate cues deposited using microfluidic devices

Lab on a Chip (2010)

Larry J. Millet Matthew E. Stewart Ralph G. Nuzzo Martha U. Gillette

Wiring the nervous system relies on the interplay of intrinsic and extrinsic signaling molecules that control neurite extension, neuronal polarity, process maturation and experience-dependent refinement. Extrinsic signals establish and enrich neuron–neuron interactions during development. Understanding how such extrinsic cues direct neurons to establish neural connections in vitro will facilitate the development of organized neural networks for investigating the development and function of nervous system networks. Producing ordered networks of neurons with defined connectivity in vitro presents special technical challenges because the results must be compliant with the biological requirements of rewiring neural networks. Here we demonstrate the ability to form stable, instructive surface-bound gradients of laminin that guide postnatal hippocampal neuron development in vitro. Our work uses a three-channel, interconnected microfluidic device that permits the production of adlayers of planar substrates through the combination of laminar flow, diffusion and physisorption. Through simple flow modifications, a variety of patterns and gradients of laminin (LN) and fluorescein isothiocyanate-conjugated poly-L-lysine (FITC–PLL) were deposited to present neurons with an instructive substratum to guide neuronal development. We present three variations in substrate design that produce distinct growth regimens for postnatal neurons in dispersed cell cultures. In the first approach, diffusion-mediated gradients of LN were formed on cover slips to guide neurons toward increasing LN concentrations. In the second approach, a combined gradient of LN and FITC–PLL was produced using aspiration-driven laminar flow to restrict neuronal growth to a 15 µm wide growth zone at the center of the two superimposed gradients. The last approach demonstrates the capacity to combine binary lines of FITC–PLL in conjunction with surface gradients of LN and bovine serum albumin (BSA) to produce substrate adlayers that provide additional levels of control over growth. This work demonstrates the advantages of spatio-temporal fluid control for patterning surface-bound gradients using a simple microfluidics-based substrate deposition procedure. We anticipate that this microfluidics-based patterning approach will provide instructive patterns and surface-bound gradients to enable a new level of control in guiding neuron development and network formation.

Laminar organization

Biological neural network

Neurite

Neural Development

10.1039/c001552k

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Citations (142)

Fourier Transform Light Scattering of Biological Structure and Dynamics

IEEE Journal of Selected Topics in Quantum Electronics (2010)

Huafeng Ding Erin Berl Zhuo Wang Larry J. Millet Martha U. Gillette

We review the principle and application of Fourier transform light scattering (FTLS), a new technique developed in our laboratory to study static and dynamic light scattering (DLS) from the biological tissues and live cells. The results demonstrate that FTLS has significant benefits over existing light scattering techniques in terms of sensitivity and resolution. We anticipate that FTLS will set the basis for disease diagnosis based on the intrinsic tissue optical properties and provide an efficient tool for quantifying cell structures and dynamics.

Spatial light modulator

10.1109/jstqe.2009.2034752

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Citations (30)

Direct Cellular Peptidomics of Supraoptic Magnocellular and Hippocampal Neurons in Low-Density Cocultures

ACS Chemical Neuroscience (2009)

Larry J. Millet Adriana Bora Jonathan V. Sweedler Martha U. Gillette

Genomic and proteomic studies of brain regions of specialized function provide evidence that communication among neurons is mediated by systems of diverse chemical messengers. These analyses are largely tissue- or population-based, whereas the actual communication is from cell-to-cell. To understand the complement of intercellular signals produced by individual neurons, new methods are required. We have developed a novel neuron-to-neuron, serum-free coculture approach that was used to determine the higher-level cellular peptidome of individual primary mammalian neurons. We isolated magnocellular neurons from the supraoptic nucleus of early postnatal rat and maintained them in serum-free low-density cultures without glial support layers; under these conditions, they required low-density cocultured neurons. Coculturing magnocellular neurons with hippocampal neurons permitted local access to individual neurons within the culture for mass spectrometry. Using direct sampling, we obtained peptide profiles for spatially distinct, identifiable neurons within the coculture. We repeatedly detected 10 peaks that we assign to previously characterized peptides and 17 peaks that remain unassigned. Peptides from the vasopressin prohormone and secretogranin-2 are attributed to magnocellular neurons, whereas neurokinin A, peptide J, and neurokinin B are attributed to cultured hippocampal neurons. This approach enables the elucidation of cell-specific prohormone processing and the discovery of cell−cell signaling peptides.

Magnocellular cell

Supraoptic nucleus

10.1021/cn9000022

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Citations (23)

Patterning the differentiation of C2C12 skeletal myoblasts

Integrative Biology (2011)

Piyush Bajaj Bobby Reddy Larry J. Millet Chunan Wei Pınar Zorlutuna

Journal Article Patterning the differentiation of C2C12 skeletal myoblasts Get access Piyush Bajaj, Piyush Bajaj Department of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA. Fax: +1 217 244-6375; Tel: +1 217 333-30972000 Micro and Nanotechnology Laboratory, University of Illinois at Urbana-Champaign, 208 North Wright Street, Urbana, IL 61801, USA Search for other works by this author on: Oxford Academic Google Scholar Bobby Reddy, Jr., Bobby Reddy, Jr. 2000 Micro and Nanotechnology Laboratory, University of Illinois at Urbana-Champaign, 208 North Wright Street, Urbana, IL 61801, USADepartment of Electrical and Computer Engineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA Search for other works by this author on: Oxford Academic Google Scholar Larry Millet, Larry Millet 2000 Micro and Nanotechnology Laboratory, University of Illinois at Urbana-Champaign, 208 North Wright Street, Urbana, IL 61801, USADepartment of Electrical and Computer Engineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA Search for other works by this author on: Oxford Academic Google Scholar Chunan Wei, Chunan Wei Department of Electrical and Computer Engineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA Search for other works by this author on: Oxford Academic Google Scholar Pinar Zorlutuna, Pinar Zorlutuna Department of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA. Fax: +1 217 244-6375; Tel: +1 217 333-30972000 Micro and Nanotechnology Laboratory, University of Illinois at Urbana-Champaign, 208 North Wright Street, Urbana, IL 61801, USANow at Harvard-MIT's Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA Search for other works by this author on: Oxford Academic Google Scholar Gang Bao, Gang Bao Department of Biomedical Engineering Georgia Institute of Technology and Emory University Atlanta GA 30332 USA Search for other works by this author on: Oxford Academic Google Scholar Rashid Bashir Rashid Bashir Department of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA. Fax: +1 217 244-6375; Tel: +1 217 333-30972000 Micro and Nanotechnology Laboratory, University of Illinois at Urbana-Champaign, 208 North Wright Street, Urbana, IL 61801, USADepartment of Electrical and Computer Engineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA E-mail: rbashir@illinois.edu Search for other works by this author on: Oxford Academic Google Scholar Integrative Biology, Volume 3, Issue 9, September 2011, Pages 897–909, https://doi.org/10.1039/c1ib00058f Published: 15 August 2011 Article history Received: 09 June 2011 Accepted: 29 July 2011 Published: 15 August 2011

Wright

C2C12

10.1039/c1ib00058f

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Citations (185)

Rapid thermal lysis of cells using silicon–diamond microcantilever heaters

Lab on a Chip (2010)

Natalya L. Privorotskaya Yi-Shao Liu Jungchul Lee Hongjun Zeng John A. Carlisle

This paper presents the design and application of microcantilever heaters for biochemical applications. Thermal lysis of biological cells was demonstrated as a specific example. The microcantilever heaters, fabricated from selectively doped single crystal silicon, provide local resistive heating with highly uniform temperature distribution across the cantilevers. Very importantly, the microcantilever heaters were coated with a layer of 100 nm thick electrically insulating ultrananocrystalline diamond (UNCD) layer used for cell immobilization on the cantilever surface. Fibroblast cells or bacterial cells were immobilized on the UNCD/cantilever surfaces and thermal lysis was demonstrated via optical fluorescence microscopy. Upon electrical heating of the cantilever structures to 93 degrees C for 30 seconds, fibroblast cell and nuclear membrane were compromised and the cells were lysed. Over 90% of viable bacteria were also lysed after 15 seconds of heating at 93 degrees C. This work demonstrates the utility of silicon-UNCD heated microcantilevers for rapid cell lysis and forms the basis for other rapid and localized temperature-regulated microbiological experiments in cantilever-based lab on chip applications.

10.1039/b923791g

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Citations (59)