We report a single cell-based rapid natural killer (NK) cell activity measurement technique that utilizes a newly developed activation stimulator cocktail (ASC) along with the lens-free shadow imaging platform. The platform, i.e., Cellytics_NK, consists of compact and low-cost optoelectronic components such as LED and CMOS image sensor, and the shadow images of photographed NK cells in a disposable cell chip are analyzed within 30 seconds with an automated algorithm. The shadow parameters such as PPD and WSM have been employed to induce a combined shadow parameter (CSP) which precisely measures and distinguishes quiescent and activated NK cells (p < 0.0001).
KEYWORDS: Bacteria, Ions, Sensors, Field effect transistors, Signal detection, Polymethylmethacrylate, Solid state electronics, Electron beam lithography, Molecules, Microfluidics
This paper presents a nanowell device that detects the nano-scale electric field fluctuations due to ion cascade in bacteria. Solid-state nano devices allow for the measurement and analysis of fluctuation on the single cell or molecule scale, which can offer orders of magnitude higher sensitivity than microscopic measurements through conventional sensors. We fabricated a nanowell that is a 150nm wide gap in the middle of a titanium line on LiNbO3 substrate. The noise in the electrical current through this gap is measured. When bacteria are infected by bacteriophage, a large amount of ions are released, which yields spatiotemporal fluctuations of electric potential captured by this nanowell. It was demonstrated that this technology can be used to identify bacteria within minutes using the high specificity of phage/bacteria interaction. The perspective of building a biochip with hundreds of nano devices, immobilized phages and microfluidic channels so as to identify a large variety of bacteria is also discussed in this paper.
In an era of potential bioterrorism and pandemics of antibiotic-resistant microbes, bacterial contaminations of food and water supplies is a major concern. There is an urgent need for the rapid, inexpensive and specific identification of bacteria under field conditions. Here we describe a method that combines the specificity and avidity of bacteriophages with fluctuation analysis of electrical noise. The method is based on the massive, transitory ion leakage that occurs at the moment of phage DNA injection into the host cell. The ion fluxes require only that the cells be physiologically viable (i.e., have energized membranes) and can occur within seconds after mixing the cells with sufficient concentrations of phage particles. To detect these fluxes, we have constructed a nano-well, a lateral, micron-size capacitor of titanium electrodes with gap size of 150 nm, and used it to measure the electrical field fluctuations in microliter (mm3) samples containing phage and bacteria. In mixtures where the analyte bacteria were sensitive to the phage, large stochastic waves with various time and amplitude scales were observed, with power spectra of approximately 1/f2 shape over at 1 - 10 Hz. Development of this SEPTIC (SEnsing of Phage-Triggered Ion Cascades) technology could provide rapid detection and identification of live, pathogenic bacteria on the scale of minutes, with unparalleled specificity. The method has a potential ultimate sensitivity of 1 bacterium/microliter (1 bacterium/mm3).
Access to the requested content is limited to institutions that have purchased or subscribe to SPIE eBooks.
You are receiving this notice because your organization may not have SPIE eBooks access.*
*Shibboleth/Open Athens users─please
sign in
to access your institution's subscriptions.
To obtain this item, you may purchase the complete book in print or electronic format on
SPIE.org.
INSTITUTIONAL Select your institution to access the SPIE Digital Library.
PERSONAL Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.