We present a novel approach, interferometric phase intensity nanoscopy, to clearly define nanometer-scale objects situated closely together, going beyond the traditional diffraction limits. Our approach capitalizes on the strengths of interferometric scattering, which we've found remarkably effective for detecting entities as minute as individual nanoparticles and proteins. We integrated multiphase analysis with enhanced sensitivity to reveal elliptical Airy patterns that directly correlate with the nanostructures we observe. A key aspect of our methodology is the use of circular polarized illumination, which is essential for differentiating nanometer-scale objects that are closely spaced and below the diffraction limit. This technique opens up new possibilities for prolonged observation of nanoscale dynamics in biology, biomedicine, and bioengineering.
Phase intensity nanoscope (PINE) is a new super-resolution method to further improve the resolution of existing techniques. PINE utilizes an integrated phase-intensity device to modulate phase differences between electric field components to distinguish nanoprobes within a diffraction-limited region. This phase-intensity separation enables continuous imaging without photobleaching. PINE achieved sub-10nm resolution of cellular structures through precise localization of populations of randomly distributed nanorod probes. The distribution of localized nanorods forms patterns of underlying structures. By defining features from probe distribution patterns and minimizing the distances of each probe to its feature projection, PINE extracts sub-10nm structural information. PINE will pave the way for new sub-10nm long-term investigations of previously unexplored material, chemical and biological dynamics.
Light-matter interactions play essential roles in realizing a new generation of nanoscale imaging to overcome traditional trade-offs between spatial resolution and time capabilities. By harnessing engineered nanophotonic materials, scattered light can be shaped to enhance light matter interactions for nanoscale bioimaging. We achieved nanoscale bioimaging with record long time capabilities. Long time nanoscopy has the potential to unlock a wide range of new dynamical studies and insights in assembly, organization and pattern formation in biological and material systems, where nanoscopic rearrangements drive group-level movements at the macroscale over time.
Bulk optical components are conventionally used to control light. However, bulk optical components are limited by optical loss and ability for miniaturization. To address this problem, we developed soft multimaterial photonic devices. The theoretical predictions and experimental results are compared with the device characterization. Plasmonic super-resolution imaging based on this device is presented as an example. We find the result of plasmonic super-resolution imaging enabled by the compact design is comparable to commercial devices.
Actin, as microfilaments in the cytoskeleton, is essential biological structure and mechanical properties for cell migration and division. These processes require new probes for visualization of actin. Fluorescent labeling as a traditional method accompanies photo bleaching and formation of free radicals that are harmful for live cells, resulting to hardly find a balance between more signals of observation and less light exposure. Here, we present new nanoparticle probes for continuous visualization of actin. We demonstrate continuous imaging of different cell division phases to reveal actin biological and mechanical properties.
The distribution and morphology of cortical actin filaments are essential in cell dynamics, such as cell division. In this work, we propose a plasmonic approach for long term imaging to study the specifications of actin dynamics in live cells. Here, we employ a new imaging setup which can acquire a set of images with spatial sparsity. Long-term and super-resolution imaging can be achieved by our imaging setup. We demonstrate examples of different morphologies of cortical actin filaments during cell division.
The method of delivery is considered to be one of the significant hurdles for CRISPR technologies. Conventional delivery of CRISPR by viral vectors are subject to their immunogenicity. Non-viral vectors methods are preferable but are limited by their biocompatibility and precision. Nano-plasmonic particles have shown great potential as noninvasive and spatiotemporally controllable vehicles for biomacromolecule delivery. Hereby, as a safe and precise way to delivery of CRISPR, we propose a remote optically manipulating method based on gold nanorods carrier. We have characterized gold nano-plasmonic carriers on single particle level by imaging spectroscopy. Then we have demonstrated the precise manipulation of individual nano plasmonic carriers on cells using optical trapping system. Lastly, the biocompatibility of the method is shown.
Cell migration is an essential cellular process that help cells to develop complex organisms, organs, and tissues, arrange cells into specialized architecture and help organize the nervous system. Nanoscale imaging has the potential to provide new insight into the mechanics of cell migration. However, quantification over time in nanoscale imaging remains difficult. Here, we visualize cell migration by non-bleaching nanoscale imaging. We present a set of quantitative metrics - length, branching, gaps, distribution, and curvature - to quantitatively analyze cell migration at the nanoscale over time.
CRISPR-Cas9 is an efficient and reliable tool to cleave targeted portions of DNA. Once the DNA been cut by the tool, genes are ready to be cut, inserted or modified as desired. Conventionally, CRISPR is carried by a virus into cells. However, potential safety issues exists, as the cell can physiologically respond to viral invasion, or its genome can be inserted with unwanted virus genes. Here, we show that gold nanoparticles serves as preferable successful vectors of CRISPR-Cas9, with extra benefit to allow stable imaging. This toxicity-free alternative method avoids the aforementioned issues and introduces the CRISPR-Cas9 complex safely
Actin dynamics plays an important role in cell activities and drives a large range of cellular process such as cell division and cell motility. To visualize the actin dynamics, we introduce non-bleaching nanoscale imaging to capture actin dynamics over long time scales. Here, we demonstrate the high resolution quantification of actin dynamics by non-bleaching nanoscale imaging for the purpose of revealing resulting mechanical properties. Through this method, we visualized and quantified actin twisting dynamics through disassembly process by actin binding proteins.
Fluorescence super-resolution imaging relies on fluorophores as contrast agents. As a result of the inevitable photobleaching and thus a limited photon budget, they put fundamental limitations on the temporal resolution. We propose non-bleaching, plasmonic nanoparticles as non-bleaching contrast agents with an unlimited photon budget such that both the temporal and spatial resolution can be pursued at the same time. We demonstrated a continuous observation of fine structural features in resolved actin networks. We believe the non-bleaching nanoscale imaging will open new doors for observing biological processes on a much longer timescales with higher spatial and temporal details.
CRISPR-Cas9 is a gene editing tool which has promise for the treatment and prevention of many complex diseases. Together with a short guide RNA, CRISPR-Cas9 can recognize and cut the corresponding DNA at a targeted location. Viruses are traditionally used as carriers of CRISPR-Cas9 into target cells. However, viruses may cause
immunogenic complications. Alternatively, gold nanoparticles can be utilized as carriers of CRISPR-Cas9.
Gold nanoparticles in the near infrared regime exhibit unique optical properties. In this work, we show gold nanoparticles can be stably and efficiently crosslinked to the CRISPR complex.These alternative carriers are toxicity free to and allow effective and efficient introduction of the CRISPR system.
High-performance UAV has been popular and in great need in recent years. The paper introduces a new method in debugging Quadrotor UAVs. Based on the infrared thermal technology and heat transfer theory, a UAV is under debugging above a hot-wire grid which is composed of 14 heated nichrome wires. And the air flow propelled by the rotating rotors has an influence on the temperature distribution of the hot-wire grid. An infrared thermal imager below observes the distribution and gets thermal images of the hot-wire grid. With the assistance of mathematic model and some experiments, the paper discusses the relationship between thermal images and the speed of rotors. By means of getting debugged UAVs into test, the standard information and thermal images can be acquired. The paper demonstrates that comparing to the standard thermal images, a UAV being debugging in the same test can draw some critical data directly or after interpolation. The results are shown in the paper and the advantages are discussed.
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