Detection ofBacillussubtilisSpores Using Peptide-Functionalized Cantilever Arrays

Dhayal, Babita; Henne, Walter A.; Doorneweerd, Derek D.; Reifenberger, R.; Low, Philip S.

doi:10.1021/ja0570887

Cited by 81 publications

(38 citation statements)

References 36 publications

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“…Recently, Low and coworkers [154] considered the detection of B. subtilis spores using resonant and static microcantilevers. For label-free detection of B. subtilis, the cantilever surface was chemically modified using a specific peptide sequence.…”

Section: Cell Detectionmentioning

confidence: 99%

Nanomechanical resonators and their applications in biological/chemical detection: Nanomechanics principles

et al. 2011

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Recent advances in nanotechnology have led to the development of nano-electro-mechanical systems (NEMS) such as nanomechanical resonators, which have recently received significant attention from the scientific community. This has not only been for their capability for the label-free detection of bio/chemical-molecules at single-molecule (or atomic) resolution for future applications such as the early diagnostics of diseases such as cancer, but also for their unprecedented ability to detect physical quantities such as molecular weight, elastic stiffness, surface stress, and surface elastic stiffness for adsorbed molecules on the surface. Most experimental works on resonator-based molecular detection have been based on the principle that molecular adsorption onto a resonator surface increases the effective mass, and consequently decreases the resonant frequencies of the nanomechanical resonator. However, this principle is insufficient to provide fundamental insights into resonator-based molecular detection at the nanoscale; this is due to recently proposed novel nanoscale detection principles including various effects such as surface effects, nonlinear oscillations, coupled resonance, and stiffness effects. Furthermore, these effects have only recently been incorporated into existing physical models for resonators, and therefore the universal physical principles governing nanoresonator-based detection have not been completely described. Therefore, our objective in this review is to overview the current attempts to understand the underlying mechanisms in nanoresonator-based detection using physical models coupled to computational simulations and/or experiments. Specifically, we will focus on issues of special relevance to the dynamic behavior of nanoresonators and their applications in biological/chemical detection: the resonance behavior of micro/nano-resonators; resonator-based chemical/biological detection; physical models of various nanoresonators such as nanowires, carbon nanotubes, and graphene. We pay particular attention to experimental and computational approaches that have been useful in elucidating the mechanisms underlying the dynamic behavior of resonators across multiple and disparate spatial/length scales, and the resulting insight into resonator-based detection that has been obtained. We additionally provide extensive discussion regarding potentially fruitful future research directions coupling experiments and simulations in order to develop a fundamental understanding of the basic physical principles that govern NEMS and NEMS-based sensing and detection applications.

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Section: Cell Detectionmentioning

confidence: 99%

Nanomechanical resonators and their applications in biological/chemical detection: Nanomechanics principles

et al. 2011

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“…16 Next, we conducted in vivo targeting experiments, where the bacteria were injected into the mouse thigh, and fluorescent probe was introduced into the bloodstream. The images in Figure 2 show the change in fluorescence intensity emanating from one mouse, out of a cohort of four, with a S. aureus NRS11 infection (∼5×10 7 Colony Forming Units in 50 μL Luria Bertani (LB) broth) in the left rear thigh. The opposite side of the mouse was injected with only the LB vehicle as a negative control.…”

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confidence: 99%

Optical Imaging of Bacterial Infection in Living Mice Using a Fluorescent Near-Infrared Molecular Probe

et al. 2006

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An optical imaging probe was synthesized by attaching a near-infrared carbocyanine fluorophore to an affinity group containing two zinc(II) dipicolylamine (Zn-DPA) units. The probe has a strong and selective affinity for the surfaces of bacteria, and it was used to image infections of Gram-positive S. aureus and Gram-negative E. coli bacteria in living nude mice. After intravenous injection, the probe selectively accumulates at the sites of localized bacterial infections in the thigh muscles of the mice.Bacterial imaging is an emerging technology that has many health and environmental applications. 1 For example, there is an obvious need to develop highly sensitive assays that can detect very small numbers of pathogenic bacterial cells in food, drinking water or biomedical samples. In other situations, the goal is to study in vivo the temporal and spatial distribution of bacteria in live animals.Optical imaging of bacteria in vivo is much less developed than methods such as radioimaging and MRI. One approach is to use bacteria that are genetically encoded to produce luciferase or green fluorescent protein. 2 A second strategy, which is the focus of this study, employs a molecular probe with a fluorescent reporter group. An obvious limitation with a live animal is restricted tissue penetration of the light. However, near-infrared (NIR) dyes with emission wavelengths in the region of 650−900 nm can propagate through two or more centimeters of tissue, and may enable deeper tissue imaging if sensitive detection techniques are employed. Molecular imaging probes can often be deconstructed into two structural components, an affinity ligand and a reporter group. In the case of bacterial targeting, previously reported affinity ligands include antibodies, 5 sugars, 6 bacteria binding peptides, 7 antimicrobial peptides, 8 enzyme substrates, 9 and antibiotic drugs. 10 Recently, we discovered that fluorescent molecular probes containing synthetic zinc(II) dipicolylamine (Zn-DPA) coordination complexes as affinity groups are able to selectively stain the surfaces of bacterial cells 11 and apoptotic animal cells. 12 Zn-DPA affinity ligands bind strongly to the anionic surfaces that are a common characteristic of these two cell-types, whereas affinity for the zwitterionic surfaces of healthy animal cells is weak. These in vitro results have motivated us to pursue in vivo studies, and we report that molecular probe 1, which has a NIR fluorophore attached to an affinity group with two Zn-DPA units, can be used for targeted, fluorescence imaging of bacterial infection in a living whole animal.The bacterial imaging probe 1 (λ max abs: 794 nm, em: 810 nm) was prepared in straightforward fashion using a carbocyanine dye as the NIR fluorophore. 13 Researchers have incorporated this fluorophore into probes for other optical imaging applications. 14 In vitro fluorescence microscopy studies proved that probe 1 can effectively stain the periphery of bacterial cells (Figure 1). In contrast, the cells are not stained when treated un...

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“…Fluorocarbon coated polymeric cantilevers have been shown to offer advantages over gold coated silicon nitride cantilevers, such as stability against temperature and pH changes (Calleja et al, 2006). Numerous biosensor systems based on surface functionalized cantilevers have been implemented with biomolecular probes for the detection of various biological targets, such as kinases and myoglobins (Arntz et al, 2003), glucose (Pei et al, 2004), bacterial cells, including Escherichia coli (Ilic et al, 2000), Listeria innocua (Gupta et al, 2004a), Bacillus subtilis spores (Dhayal et al, 2006), vaccinia virus particles (Johnson et al, 2006), RNA (Zhang et al, 2006), and cancer markers (Wu et al, 2001). …”

Section: Microcantileversmentioning

confidence: 99%

Micro- and nanodevices integrated with biomolecular probes

Alapan

İçöz

Gürkan

2015

Biotechnology Advances

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Understanding how biomolecules, proteins and cells interact with their surroundings and other biological entities has become the fundamental design criterion for most biomedical micro- and nanodevices. Advances in biology, medicine, and nanofabrication technologies complement each other and allow us to engineer new tools based on biomolecules utilized as probes. Engineered micro/nanosystems and biomolecules in nature have remarkably robust compatibility in terms of function, size, and physical properties. This article presents the state of the art in micro- and nanoscale devices designed and fabricated with biomolecular probes as their vital constituents. General design and fabrication concepts are presented and three major platform technologies are highlighted: microcantilevers, micro/nanopillars, and microfluidics. Overview of each technology, typical fabrication details, and application areas are presented by emphasizing significant achievements, current challenges, and future opportunities.

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Detection ofBacillussubtilisSpores Using Peptide-Functionalized Cantilever Arrays

Cited by 81 publications

References 36 publications

Nanomechanical resonators and their applications in biological/chemical detection: Nanomechanics principles

Nanomechanical resonators and their applications in biological/chemical detection: Nanomechanics principles

Optical Imaging of Bacterial Infection in Living Mice Using a Fluorescent Near-Infrared Molecular Probe

Micro- and nanodevices integrated with biomolecular probes

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