A printed nanolitre-scale bacterial sensor array (original) (raw)

Printing Multistrain Bacterial Patterns with a Piezoelectric Inkjet Printer

PLoS ONE, 2007

Many studies involving interacting microorganisms would benefit from simple devices able to deposit cells in precisely defined patterns. We describe an inexpensive bacterial piezoelectric inkjet printer (adapted from the design of the POSaM oligonucleotide microarrayer) that can be used to ''print out'' different strains of bacteria or chemicals in small droplets onto a flat surface at high resolution. The capabilities of this device are demonstrated by printing ordered arrays comprising two bacterial strains labeled with different fluorescent proteins. We also characterized several properties of this piezoelectric printer, such as the droplet volume (of the order of tens of pl), the distribution of number of cells in each droplet, and the dependence of droplet volume on printing frequency. We established the limits of the printing resolution, and determined that the printed viability of Escherichia coli exceeded 98.5%.

Biological laser printing of genetically modified Escherichia coli for biosensor applications

Biosensors and Bioelectronics, 2004

One of the primary requirements of cell-or tissue-based sensors is the placement of cells and cellular material at or near the sensing elements of the device. The ability to achieve precise, reproducible and rapid placement of cells is the focus of this study. We have developed a technique, biological laser printing or BioLP TM , which satisfies these requirements and has advantages over current technologies. BioLP TM is capable of rapidly depositing patterns of active biomolecules and living cells onto a variety of material surfaces. Unlike ink jet or manual spotting techniques, this process delivers small volume (nl to fl) aliquots of biomaterials without the use of an orifice, thus eliminating potential clogging issues and enabling diverse classes of biomaterials to be deposited. This report describes the use of this laser-based printing method to transfer genetically-modified bacteria capable of responding to various chemical stressors onto agar-coated slides and into microtiter plates. The BioLP TM technology enables smaller spot sizes, increased resolution, and improved reproducibility compared to related technologies.

Microbial sensor cell arrays

Current Opinion in Biotechnology, 2012

Motivated by the advantages endowed by high-throughput analysis, researchers have succeeded in incorporating multiple reporter cells into a single platform; the technology now allows the simultaneous scrutiny of a large collection of sensor strains. We review current aspects in cell array technology with emphasis on microbial sensor arrays. We consider various techniques for patterning live cells on solid surfaces, describe different array-based applications and devices, and highlight recent efforts for live cell storage. We review mathematical approaches for deciphering the data emanating from bioreporter collections, and discuss the future of single cell arrays. Innovative technologies for cell patterning, preservation and interpretation are continuously being developed; when they all mature, cell arrays may become an efficient analytical tool, in a scope resembling that of DNA microarray biochips.

A Low-Cost, 3D-Printed Biosensor for Rapid Detection of Escherichia coli

2022

Detection of bacterial pathogens is significant in the fields of food safety, medicine, public health, etc. If bacterial pathogens are not treated promptly, antimicrobial resistance is possible and can lead to morbidity and mortality. Current bacterial detection methodologies rely on laboratory-based techniques that pose limitations such as long turnaround detection times, expensive costs, inadequate accuracy, and required trained specialists. Here, we describe a cost-effective and portable 3D-printed electrochemical biosensor that facilitates rapid detection of certain Escherichia coli (E. coli) strains (DH5α, BL21, TOP10, and JM109) within 15 minutes using 500 μL of sample and costs $2.50 per test. The sensor displayed an excellent limit of detection (LOD) of 53 cfu, limit of quantification (LOQ) of 270 cfu, and showed cross-reactivity with strains BL21 and JM109 due to shared epitopes. This advantageous diagnostic device is a potential candidate for high-frequency testing at poin...

Cell bioprinting as a potential high-throughput method for fabricating cell-based biosensors (CBBs)

2009 IEEE Sensors, 2009

Cell-based biosensors (CBBs) are becoming an important tool for biosecurity applications and rapid diagnostics. For current CBBs technology, cell immobilization and high throughput fabrication are the main challenges. To address these in this study, the feasibility of bioprinting cell-laden hydrogel to fabricate CBBs at high throughput was investigated and cell response was tracked by using lensless charge-coupled device (CCD) technology. This study indicated that (i) a cellladen collagen printing platform was capable of immobilizing cells (smooth muscle cells) in collagen droplets with precise spatial control and pattern them onto surfaces, (ii) high postprinting cell viability was achieved (>94%) and the immobilized cells proliferated over five days, (iii) the immobilized cells maintain their biological and physiological sensitivity to environmental stimuli (e.g. environmental temperature change and lysis by adding of de-ionized water), as quantified by change in cell spread size (decreasing from ~3000 μm 2 at t=0 hour to ~600 μm 2 I.

Microbial Nanocellulose Printed Circuit Boards for Medical Sensing

Sensors, 2020

We demonstrate the viability of using ultra-thin sheets of microbially grown nanocellulose to build functional medical sensors. Microbially grown nanocellulose is an interesting alternative to plastics, as it is hydrophilic, biocompatible, porous, and hydrogen bonding, thereby allowing the potential development of new application routes. Exploiting the distinguishing properties of this material enables us to develop solution-based processes to create nanocellulose printed circuit boards, allowing a variety of electronics to be mounted onto our nanocellulose. As proofs of concept, we have demonstrated applications in medical sensing such as heart rate monitoring and temperature sensing—potential applications fitting the wide-ranging paradigm of a future where the Internet of Things is dominant.

Arrays of nanoelectromechanical biosensors functionalized by microcontact printing

Nanotechnology, 2012

The biofunctionalization of nanoelectromechanical structures is critical for the development of new classes of biosensors displaying improved performances and higher-level of integration. We propose a modified microcontact printing method for the functionalization and passivation of large arrays of nanocantilevers in a single, self-aligned step. Using fluorescence microscopy and resonant frequency measurements, we demonstrate (1) the bioactivity and the anti-fouling property of deposited antibodies and BSA molecules and (2) the preservation of the nanostructures' mechanical integrity.

Controlled assembly of bacteria on chemical patterns using soft lithography

Colloids and Surfaces B: Biointerfaces, 2008

Highly ordered arrays of single living bacteria were obtained by selective adsorption of bacteria onto chemical patterns with micrometric resolution. The chemically engineered template surfaces were prepared with the combination of microcontact printing process and a simple incubation technique. This methodology can be used for fundamental studies of bacterium's inner mechanisms and sub-cellular organization as well as for interfacing living bacteria with artificial microsystems.

Electronically directed integration of whole-cell biosensors on bio-chips

ECS Transactions, 2010

This paper presents a whole-cell bio-chip system where viable, functioning cells are deposited onto solid surfaces that are a part of a micro-machined system. The development of such novel hybrid functional sensors depends on the cell deposition methods; in this work new approach integrating live bacterial cells on a bio-chip using electrophoretic deposition is presented. The bio-material deposition technique was characterized under various driving potential and chamber configurations. The deposited bio-mass included genetically engineered bacterial cells generating electrochemically active byproduct upon exposure to toxic materials in the aqueous solution. In this paper we present the deposition apparatus and methods, as well as the characterization results, e.g. signal vs. time and induction factor, of such chips and discussing the highlight and problems of the new deposition method.

Redesigning of Microbial Cell Surface and Its Application to Whole-Cell Biocatalysis and Biosensors

Applied Biochemistry and Biotechnology, 2017

Microbial cell surface display technology can redesign cell surfaces with functional proteins and peptides to endow cells some unique features. Foreign peptides or proteins are transported out of cells and immobilized on cell surface by fusing with anchoring proteins, which is an effective solution to avoid substance transfer limitation, enzyme purification, and enzyme instability. As the most frequently used prokaryotic and eukaryotic protein surface display system, bacterial and yeast surface display systems have been widely applied in vaccine, biocatalysis, biosensor, bioadsorption, and polypeptide library screening. In this review of bacterial and yeast surface display systems, different cell surface display mechanisms and their applications in biocatalysis as well as biosensors are described with their strengths and shortcomings. In addition to single enzyme display systems, multi-enzyme co-display systems are presented here. Finally, future developments based on our and other previous reports are discussed.