Christopher N. LaFratta

Multiplexed Sandwich Immunoassays using Electrochemiluminescence Imaging Resolved at the Single Bead Level

A new class of bead-based microarray that uses electrogenerated chemiluminescence (ECL) as a readout mechanism to detect multiple antigens simultaneously is presented. This platform demonstrates the possibility of performing highly multiplexed assays using ECL because all the individual sensing beads in the array are simultaneously imaged and individually resolved by ECL. Duplex and triplex assay results are demonstrated as well as a cross reactivity study.

Acrylic-based resin with favorable properties for three-dimensional two-photon polymerization

We describe an acrylic-based prepolymer resin that is ideally suited for the fabrication of three-dimensional structures with two-photon polymerization. We characterize the photochemical and photophysical properties of the photoinitiator and present representative structures that demonstrate the favorable mechanical and optical properties of the polymer.

Multiphoton laser direct writing of two-dimensional silver structures.

We report a novel and efficient method for the laser direct writing of two-dimensional silver structures. Multiphoton absorption of a small fraction of the output of a Ti:sapphire oscillator is sufficient to photoreduce silver nitrate in a thin film of polyvinylpyrrolidone that has been spin-coated on a substrate. The polymer can then be washed away, leaving a pattern consisting of highly interconnected silver nanoparticles. We report the characterization of the silver patterns using scanning electron and atomic force microscopies, and demonstrate the application of this technique in the creation of diffraction gratings.

Polymer microcantilevers fabricated via multiphoton absorption polymerization

We have used multiphoton absorption polymerization to fabricate a series of microscale polymer cantilevers. Atomic force microscopy has been used to characterize the mechanical properties of microcantilevers with spring constants that were found to span more than four decades. From these data, we extracted a Young’s modulus of E=0.44GPa for these microscale cantilevers. The wide stiffness range and relatively low elastic modulus of the microstructures make them attractive candidates for a range of microcantilever applications, including measurements on soft matter.

CMOS Microelectrode Array for Electrochemical Lab-on-a-Chip Applications

Microelectrode arrays (MEAs) offer numerous benefits over macroelectrodes due to their smaller sample size requirement, small form factor, low-power consumption, and higher sensitivity due to increased rates of mass transport. These features make MEAs well suited for microfluidic lab-on-a-chip applications. This paper presents two implementations of MEAs with and without an on chip potentiostat. We first describe an 8times8 array of 6 mum circular microelectrodes with center to center 37 mum spacing fabricated on silicon using conventional microfabrication techniques. Pads are provided for external connections to a potentiostat for electrochemical analysis. The second implementation is an individually addressable 32times32 array of 7 mum square microelectrodes with 37 mum center to center spacing on a CMOS chip with built-in very-large-scale integration potentiostat for electrochemical analysis. The integrated CMOS MEA is post processed at the die level to coat the exposed Al layers with Au. To verify microelectrode array behavior with individual addressability, cyclic voltammetry was performed using a potassium ferricyanide (K3Fe(CN)6) solution.

Infochemistry and infofuses for the chemical storage and transmission of coded information

This article describes a self-powered system that uses chemical reactions—the thermal excitation of alkali metals—to transmit coded alphanumeric information. The transmitter (an “infofuse”) is a strip of the flammable polymer nitrocellulose patterned with alkali metal ions; this pattern encodes the information. The wavelengths of 2 consecutive pulses of light represent each alphanumeric character. While burning, infofuses transmit a sequence of pulses (at 5–20 Hz) of atomic emission that correspond to the sequence of metallic salts (and therefore to the encoded information). This system combines information technology and chemical reactions into a new area—“infochemistry”—that is the first step toward systems that combine sensing and transduction of chemical signals with multicolor transmission of alphanumeric information.

InfoBiology by printed arrays of microorganism colonies for timed and on-demand release of messages

This paper presents a proof-of-principle method, called InfoBiology, to write and encode data using arrays of genetically engineered strains of Escherichia coli with fluorescent proteins (FPs) as phenotypic markers. In InfoBiology, we encode, send, and release information using living organisms as carriers of data. Genetically engineered systems offer exquisite control of both genotype and phenotype. Living systems also offer the possibility for timed release of information as phenotypic features can take hours or days to develop. We use growth media and chemically induced gene expression as cipher keys or “biociphers” to develop encoded messages. The messages, called Steganography by Printed Arrays of Microbes (SPAM), consist of a matrix of spots generated by seven strains of E. coli, with each strain expressing a different FP. The coding scheme for these arrays relies on strings of paired, septenary digits, where each pair represents an alphanumeric character. In addition, the photophysical properties of the FPs offer another method for ciphering messages. Unique combinations of excited and emitted wavelengths generate distinct fluorescent patterns from the Steganography by Printed Arrays of Microbes (SPAM). This paper shows a new form of steganography based on information from engineered living systems. The combination of bio- and “photociphers” along with controlled timed-release exemplify the capabilities of InfoBiology, which could enable biometrics, communication through compromised channels, easy-to-read barcoding of biological products, or provide a deterrent to counterfeiting.

Dynamic microbead arrays for biosensing applications

In this paper we present the development of an optical tweezers platform capable of creating on-demand dynamic microbead arrays for the multiplexed detection of biomolecules. We demonstrate the use of time-shared optical tweezers to dynamically assemble arrays of sensing microspheres, while simultaneously recording fluorescence signals in real time. The detection system is able to achieve multiplexing by using quantum dot nanocrystals as both signaling probes and encoding labels on the surface of the trapped microbeads. The encoding can be further extended by using a range of bead sizes. Finally, the platform is used to detect and identify three genes expressed by pathogenic strains of Escherichia coli O157:H7. The in situ actuation enabled by the optical tweezers, combined with multiplexed fluorescence detection offers a new tool, readily adaptable to biosensing applications in microfluidic devices, and could potentially enable the development of on-demand diagnostics platforms.

Two-Photon Polymerization Metrology: Characterization Methods of Mechanisms and Microstructures

The ability to create complex three-dimensional microstructures has reached an unprecedented level of sophistication in the last 15 years. For the most part, this is the result of a steady development of the additive manufacturing technique named two-photon polymerization (TPP). In a short amount of time, TPP has gone from being a microfabrication novelty employed largely by laser specialists to a useful tool in the hands of scientists and engineers working in a wide range of research fields including microfluidics. When used in combination with traditional microfabrication processes, TPP can be employed to add unique three-dimensional components to planar platforms, thus enabling the realization of lab-on-a-chip solutions otherwise impossible to create. To take full advantage of TPP, an in-depth understanding is required of the materials photochemistry and the fabricated microstructures’ mechanical and chemical properties. Thus, we review methods developed so far to investigate the underling mechanism involved during TPP and analytical methods employed to characterize TPP microstructures. Furthermore, we will discuss potential opportunities for using optofluidics and lab-on-a-chip systems for TPP metrology.

Optical tweezers for medical diagnostics

Laser trapping by optical tweezers makes possible the spectroscopic analysis of single cells. Use of optical tweezers in conjunction with Raman spectroscopy has allowed cells to be identified as either healthy or cancerous. This combined technique is known as laser tweezers Raman spectroscopy (LTRS), or Raman tweezers. The Raman spectra of cells are complex, since the technique probes nucleic acids, proteins, and lipids; but statistical analysis of these spectra makes possible differentiation of different classes of cells. In this article the recent development of LTRS is described along with two illustrative examples for potential application in cancer diagnostics. Techniques to expand the uses of LTRS and to improve the speed of LTRS are also suggested.