The goal of this work is to develop in vivo photoacoustic (PA) flow cytometry (PAFC) for time-resolved detection of circulating absorbing objects, either without labeling or with nanoparticles as PA labels. This study represents the first attempt, to our knowledge, to demonstrate the capability of PAFC with tunable near-infrared (NIR) pulse lasers for real-time monitoring of gold nanorods, Staphylococcus aureus and Escherichia coli labeled with carbon nanotubes (CNTs), and contrast dye Lymphazurin in the microvessels of mouse and rat ears and mesenteries. PAFC shows the unprecedented threshold sensitivity in vivo as one gold nanoparticle in the irradiated volume and as one bacterium in the background of 108 of normal blood cells. The CNTs are demonstrated to serve as excellent new NIR high-PA contrast agents. Fast Lymphazurin diffusion in live tissue is observed with rapid blue coloring of a whole animal body. The enhancement of the thermal and acoustic effects is obtained with clustered, multilayer, and exploded nanoparticles.This novel combination of PA microscopy/spectroscopy and flow cytometry may be considered as a new powerful tool in biological research with the potential of quick translation to humans, providing ultrasensitive diagnostics of pathogens (e.g., bacteria, viruses, fungi, protozoa, parasites, helminthes), metastatic, infected, inflamed, stem, and dendritic cells, and pharmacokinetics of drug, liposomes, and nanoparticles in deep vessels (with focused transducers) among other potential applications.
Effective and rapid detection of nitroaromatic explosive compounds, especially trinitrotoluene (TNT), is very important to homeland security as well as to environmental monitoring of contaminants in soil and water, and landmine detection. In this research, we explore a novel nanoscale flagellar motor based TNT detection system (nFMTNT). The nFMTNT is a bio-hybrid MEMS system which combines genetically engineered flagellar motors and MEMS devices. The system consists of three major components: (1) a non-pathogenic, genetically modified Escherichia coli strain KAF95 with a rotating flagellar filament, (2) a microchannel with tethered cells, and (3) a sub-micron bead attached to a rotating flagellar filament. The operational principle of nFMTNT is based on detecting the change in the rotational behavior of the nanoscale flagellar filament in the presence of TNT. The rotational behavior of flagellar filaments of E. coli KAF95 was shown to be extremely sensitive to the presence of nitrate or nitrite. Normally, the flagellar filaments were locked in to rotate in the counterclockwise direction. However, when a nitrate or nitrite was present in the immediate environment, the filaments cease to rotate. Our results indicate that the threshold concentrations required for this response were 10-4 M for nitrate and 10-3 M for nitrite. This is equivalent to around 10 pg of nitrate and 100 pg of nitrite, based on the dimension of the MEMS-based reaction system used for the experiment (400 μm × 100 μm × 40 μm). These detection limits can be even lower when the size of the system is reduced.
Conference Committee Involvement (1)
Micro (MEMS) and Nanotechnologies for Defense and Security