Bioimaging Systems Research Articles

Comparing the Rod-Like and Spherical BODIPY Nanoparticles in Cellular Imaging.

To design efficient nanoparticles for bioimaging, it is necessary to obtain nanoparticles with desired cellular uptake and biofunction. There are many studies have shown that cellular uptake largely depends on the geometric properties of nanoparticles. In this work, the organic nanoparticles with rod-like and spherical shapes were fabricated, and their cellular behaviors were studied and compared in detail via cellular uptake and bioimaging effect. The nanoparticles with spherical and rod-like morphology both can be internalized by HeLa and HepG2 cells, but the rod-like nanoparticles showed better imaging performance than their spherical counterpart. Above results presented that the rod-like nanoparticles possess great potential for bioimaging in efficient delivery and ideal imaging efficacy. Our studies may provide useful and fundamental information for designing efficient bioimaging systems.

SERS Substrate with Silk Nanoribbons as Interlayer Template.

The formation of hot spots is an effective approach to improve the performance of surface-enhanced Raman scattering (SERS). Silk nanoribbons (SNRs), with a height of about 1-2 nm, and Au nanoparticles (AuNPs) were assembled by electrostatic interactions to introduce sandwich hot spot structures. These sandwich structures were optimized by tuning the ratio of SNRs and AuNPs, resulting in strong SERS signals with a sensitivity of 10-13 M and enhancement factor (EF) of 5.8 × 106. Improved SERS spectrum uniformity with relative standard deviation (RSD) about 11.2% was also achieved due to the homogeneous distribution of these hot spot structures. The inherent biocompatibility of SNRs and facile fabrication processes utilized endowed the SERS substrates significant benefits toward biomedical applications, confirmed by cytocompatibility and improved SERS bioimaging capacity in vitro. The results of this study suggest the feasibility of forming high performance bioimaging systems through the use of naturally derived materials with special nanostructures.

Sample-Efficient Algorithms for Recovering Structured Signals From Magnitude-Only Measurements

We consider the problem of recovering a signal $ \mathbf {x^{*}}\in \mathbb {R}^{n}$ , from magnitude-only measurements, $y_{i} = \left |{ \left \langle{ { \mathbf {a}_{i}}, \mathbf {x^{*}} }\right \rangle }\right | $ for $i=\{1,2,\ldots ,m\}$ . This is a stylized version of the classical phase retrieval problem , and is a fundamental challenge in nano- and bio-imaging systems, astronomical imaging, and speech processing. It is well-known that the above problem is ill-posed, and therefore some additional assumptions on the signal and/or the measurements are necessary. In this paper, we consider the case where the underlying signal $ \mathbf {x^{*}}$ is $s$ -sparse. For this case, we develop a novel recovery algorithm that we call Compressive Phase Retrieval with Alternating Minimization , or CoPRAM . Our algorithm is simple and is obtained via a natural combination of the classical alternating minimization approach for phase retrieval with the CoSaMP algorithm for sparse recovery. Despite its simplicity, we prove that our algorithm achieves a sample complexity of $ \mathcal {O}\left ({{s^{2} \log n}}\right )$ with Gaussian measurements $ { \mathbf {a}_{i}}$ , which matches the best known existing results; moreover, it also demonstrates linear convergence in theory and practice. An appealing feature of our algorithm is that it requires no extra tuning parameters other than the signal sparsity level $s$ . Moreover, we show that our algorithm is robust to noise. The quadratic dependence of sample complexity on the sparsity level is sub-optimal, and we demonstrate how to alleviate this via additional assumptions beyond sparsity. First, we study the (practically) relevant case where the sorted coefficients of the underlying sparse signal exhibit a power law decay. In this scenario, we show that the CoPRAM algorithm achieves a sample complexity of $ \mathcal {O}\left ({{s \log n}}\right )$ , which is close to the information-theoretic limit. We then consider the case where the underlying signal $ \mathbf {x^{*}}$ arises from structured sparsity models. We specifically examine the case of block-sparse signals with uniform block size of $b$ and block sparsity $k=s/b$ . For this problem, we design a recovery algorithm that we call Block CoPRAM that further reduces the sample complexity to $ \mathcal {O}\left ({{ks \log n}}\right )$ . For sufficiently large block lengths of $b=\Theta (s)$ , this bound equates to $ \mathcal {O}\left ({{s \log n}}\right )$ . To our knowledge, our approach constitutes the first family of linearly convergent algorithms for signal recovery from magnitude-only Gaussian measurements that exhibit a sub-quadratic dependence on the signal sparsity level.

Enabling magnetic resonance imaging of hollow-core microstructured optical fibers via nanocomposite coating.

Optical fibers are widely used in bioimaging systems as flexible endoscopes that are capable of low-invasive penetration inside hollow tissue cavities. Here, we report on the technique that allows magnetic resonance imaging (MRI) of hollow-core microstructured fibers (HC-MFs), which paves the way for combing MRI and optical bioimaging. Our approach is based on layer-by-layer assembly of oppositely charged polyelectrolytes and magnetite nanoparticles on the inner core surface of HC-MFs. Incorporation of magnetite nanoparticles into polyelectrolyte layers renders HC-MFs visible for MRI and induces the red-shift in their transmission spectra. Specifically, the transmission shifts up to 60 nm have been revealed for the several-layers composite coating, along with the high-quality contrast of HC-MFs in MRI scans. Our results shed light on marrying fiber-based endoscopy with MRI to open novel possibilities for minimally invasive clinical diagnostics and surgical procedures in vivo.

Self-Assembly of Amphiphilic Peptides for Recognizing High Furin-Expressing Cancer Cells.

Self-assembled nanostructures of amphiphilic peptides have a wide range of applications in bioimaging and delivery systems. In this study, we design and synthesize a biocompatible amphiphilic peptide (C-3) consisting of an RVRRFFF sequence and a nitrobenzoxadiazole fluorophore that can self-assemble into stable micelles for specifically detecting furin, a kind of proprotein convertase with promoting tumor progression. The self-assembly of C-3 with a β-sheet nanostructure is capable of a rapid and specific response to furin in only 5 min in aqueous solution because of the existence of the RVRR motif in the C-3 molecule. The C-3 nanostructures thus can selectively distinguish high furin-expressing cancer cells, like MDA-MB-231 cells, a kind of human breast cancer cells, from normal cells. Furthermore, the C-3 self-assembly can stay in living cells for a long time and are capable of durable detection of intracellular furin, being good for tracer analysis. To our knowledge, this is the first example of self-assembly of a soluble amphiphilic peptide that can selectively detect furin in high furin-expressing live cells.

Single-Irradiation Simultaneous Dual-Modal Bioimaging Using Nanostructure Scintillators as Single Contrast Agent.

The rising demand for clinical diagnosis tools has led to extensive research on multimodal bioimaging systems. Unlike single-modal detection, multimodal imaging not only can provide both function and structure information but also can address the issue of sensitivity, depth, and cost. Despite enormous efforts, conventional step-by-step procedures for obtaining multimodal imaging pose a significant constraint on their practical applications. In this work, X-rays as highly penetrating radiation is proposed as a single-irradiation resource, while lanthanide-based nanostructure scintillators are employed as the single contrast agent to attenuate and convert X-rays, achieving computer tomography (CT) and optical dual-modal imaging at the same time. In other words, CT and optical dual-modal imaging are simultaneously produced via single radiation combined with single contrast agent. The function and structure information of targeted tumors in a mouse model can be clearly provided with large penetration and high sensitivity, indicating that this strategy is a simple but promising route for multimodal imaging of molecular disease and preclinical applications.

Laser printed flat lenses for optofluidics

Conventional three-dimensional optics requires curvature to control the wave front of light thus making it difficult to reduce the size of the optical systems. Furthermore, for correction of optical aberrations, complex optical systems comprising more than one lens are used. This adds additional bulk, mass and complexity to the optical systems. Recent development in diffractive optics has enabled new thin lightweight optical elements such as metalenses. We introduce resonant laser printing technique as a flexible photo-thermal technology for metalens fabrication with the ability to control the light with microscale precision. Our laser printed metalenses can be integrated in bio-sensors, bio-imaging systems, and optofluidical devices.

Preparation of calcium phosphate nanoparticles hybridized with europium(III) complex for novel luminescent organic-inorganic systems

We have synthesized novel inorganic–organic hybrid systems based on the interactions between calcium phosphate (CP) and tris(2,2,6,6-tetramethyl-3,5-heptanedionato)europium (III) (EuTH) complex to clarify the states of the EuTH molecules. We achieved the nucleation and crystal growth of electronically localized of EuTH molecules using two different synthetic processes (based on the timing of the addition EuTH before and after the CP nucleation stage). The coordination environment of the Eu3+ ion inside/on the hybrids was clearly changed, having the higher spatial symmetry states in the hybrid. The aspect ratios of the hybrid crystallites were 2.5–6.0, suggesting that the EuTH molecules selectively interacted with the a–plane of the CP nuclei to induce preferential crystal growth along with the uncovered c–plane. The segregated EuTH molecules on CP surfaces suppressed Ca2+ dissolution from the hybrids in phosphate buffer saline. In the excitation spectra of the hybrids, the 7F0 → 5L6 transition appeared as a result of the interaction between the EuTH and phosphate ions, suggesting that the phosphate ions affected the coordination environment. In the luminescence spectra of the hybrids, the luminescence intensity ratio of the magnetic dipole transition (5D0 → 7F1) to electric dipole transition (5D0 → 7F2) were higher than those of EuTH and the other Eu3+-doped inorganic systems. Thus, the higher spatial symmetry states of the Eu3+ ion in the hybrids resulted in greater luminescence intensities and quantum efficiencies, which would be affected by the phosphorus anti-quenching. In the future, CP hybrids with various organic complexes will be prepared, and these could be applied in the biomedical fields such as in bio-imaging and drug delivery systems.

OpenHiCAMM: High-Content Screening Software for Complex Microscope Imaging Workflows

SummaryHigh-content image acquisition is generally limited to cells grown in culture, requiring complex hardware and preset imaging modalities. Here we report an open source software package, OpenHiCAMM (Open Hi Content Acquisition for μManager), that provides a flexible framework for integration of generic microscope-associated robotics and image processing with sequential workflows. As an example, we imaged Drosophila embryos, detecting the embryos at low resolution, followed by re-imaging the detected embryos at high resolution, suitable for computational analysis and screening. The OpenHiCAMM package is easy to use and adapt for automating complex microscope image tasks. It expands our abilities for high-throughput image-based screens to a new range of biological samples, such as organoids, and will provide a foundation for bioimaging systems biology.

Microwave‐Assisted Synthesis of Biocompatible Silk Fibroin‐Based Carbon Quantum Dots

AbstractA biocompatible silk fibroin‐based carbon quantum dot (SF‐CQD) is first synthesized under microwave irradiation for a short time. This fast and environmentally safe technique produce well‐defined nanosized SF‐CQDs. The SF‐CQDs have good crystallinity, a strong emission peak in the blue‐color region, high quantum yield, and the potential for modification with various functional groups on the surface. These SF‐CQDs demonstrate stable emission, good water dispersity, low toxicity, and good biocompatibility. These properties show the great potential of these SF‐CQDs for use in biomedical applications including bioimaging, biosensing, and drug delivery systems.

Electrochemicolor Imaging Using an LSI-Based Device for Multiplexed Cell Assays.

Multiplexed bioimaging systems have triggered the development of effective assays, contributing new biological information. Although electrochemical imaging is beneficial for quantitative analysis in real time, monitoring multiple cell functions is difficult. We have developed a novel electrochemical imaging system, herein, using a large-scale integration (LSI)-based amperometric device for detecting multiple biomolecules simultaneously. This system is designated as an electrochemicolor imaging system in which the current signals from two different types of biomolecules are depicted as a multicolor electrochemical image. The mode-selectable function of the 400-electrode device enables the imaging system and two different potentials can be independently applied to the selected electrodes. The imaging system is successfully applied for detecting multiple cell functions of the embryonic stem (ES) cell and the rat pheochromocytoma (PC12) cell aggregates. To the best of our knowledge, this is the first time that a real-time electrochemical mapping technique for multiple electroactive species, simultaneously, has been reported. The imaging system is a promising bioanalytical method for exploring complex biological phenomena.

Two-Level Shape Changes of Polymeric Microcuboids Prepared from Crystallizable Copolymer Networks

Polymeric microdevices bearing features like nonspherical shapes or spatially segregated surface properties are of increasing importance in biological and medical analysis, drug delivery, and bioimaging or microfluidic systems as well as in micromechanics, sensors, information storage, or data carrier devices. Here, a method to fabricate programmable microcuboids with shape-memory capability and the quantification of their recovery at different levels is reported. The method uses the soft lithographic technique to create microcuboids with well-defined sizes and surface properties. Microcuboids having an edge length of 25 μm and a height of 10 μm were prepared from cross-linked poly[ethylene-co-(vinyl acetate)] (cPEVA) with different vinyl acetate contents and were programmed by compression with various deformation degrees at elevated temperatures. The microlevel shape-recovery of the cuboidal geometry during heating was monitored by optical microscopy (OM) and atomic force microscopy (AFM) studying the re...

Quantitation of Suicidal Netosis Using Convolutional Neural Networks in Human Neutrophils

Suicidal NETosis (Neutrophil Extracellular Traps) is a mode of cell death involving de-condensation of chromatin and fusion with cytoplasmic granule proteins to create a net-like structure for entrapping pathogenic elements. Despite its name, NETosis does not occur exclusively in neutrophils but rather in all myeloid cells of the immune system except dendritic cells. A growing list of diseases and conditions implicate NETs in their etiology and the progression of symptoms. These conditions include sepsis, cystic fibrosis, lupus nephritis, acute respiratory distress syndrome (ARDS) and deep vein thrombosis. There are several different quantitative methods to assess NETosis making comparative analysis across the field of NETosis research extremely challenging. To further define NETosis and facilitate new discoveries in this field, we have developed a novel assay that combines high-content microscopy with Convolutional Neural Networks (CNN). CNN is the revolutionary image recognition technology used in popular social media facial features recognition systems to 1) quantify the percentage of NETotic and non-NETotic nuclei in a cell population, 2) classify NETotic, necrotic and normal nuclei in a mixed population of neutrophils and 3) differentiate between two different NETotic nuclei generated by two of the most commonly used agonists in NETosis research, the PKC activator PMA and A23187, a Ca+2 ionophore. Furthermore we have optimized neutrophil isolation and culture conditions to yield reproducible results with low inter-and intra-assay variability. Image acquisition was performed on adherent neutrophils using the automated BD Pathway Bio-imaging Systems with autofocusing function that allows imaging of approximately 10% of a well in a 96-well plate. An algorithm was created for image segmentation and recognition of NETotic nuclei based on size, DNA staining intensity and pattern. Data generated by this semi-automated analysis system were used to train a CNN to count NETotic and non-NETotic nuclei. These methods allowed us to calculate the EC50 for PMA (2.2 nM) and A23187 (2.5 uM) and to differentiate between their distinct mechanisms of NETosis. CNNs are able to classify NETotic phenotypes, without the use of expensive antibodies or reagents. Our assay represents a crucial first step for standardizing and quantitating NETosis, which will allow better data interpretation across different labs and provide a potential screening tool for known and as yet undiscovered NETosis agonists. DisclosuresNo relevant conflicts of interest to declare.

Anion-induced emissive nanoparticles for tunable fluorescence detection of pyrophosphate and bioimaging application

Nanostructures based on molecular assemblies are emerging with promising applications in biosensing, bioimaging and light-harvesting systems. In this study, three terpyridine-based Zn2+ complexes have been prepared and utilized as fluorescent probes for selective detection of pyrophosphate (PPi) over ATP and ADP. These complexes exhibited differential fluorescence responses upon addition of PPi. Both turn-on and ratiometric probes were successfully developed. More importantly, PPi-induced emissive nanoparticles were clearly observed when these terpyridine-based Zn2+ probes were treated with PPi. All probes have been employed for cell imaging and nuclei of cells are lighted up.

Pluronic mimicking fluorescent carbon nanoparticles conjugated with doxorubicin via acid-cleavable linkage for tumor-targeted drug delivery and bioimaging

Fluorescent carbon nanoparticles based target drug delivery with bio-imaging systems offer tremendous scope for future. We described Pluronic mimicking fluorescent nanoparticles (Plu-FNPs) surface decorated doxorubicin (DOX) via acid-cleavable hydrazone linkages with tumor target folic acid (FA) [Plu-FNPs-FA-DOX]. The acid labile of hydrazine linkage to DOX can easily break off by controlled pH inducing release of conjugated DOX at site of over-expressed folate receptors (FR). This nanoparticle system can deliver DOX to the target FR and trace the delivery pathway of cancer cells. The approaching platform demonstrates the selectivity and sensitivity of molecular targets, thereby able to maximize the therapeutic efficiency.

Functionalized Nanolipobubbles Embedded Within a Nanocomposite Hydrogel: a Molecular Bio-imaging and Biomechanical Analysis of the System

The purpose of this study was to explore the use of molecular bio-imaging systems and biomechanical dynamics to elucidate the fate of a nanocomposite hydrogel system prepared by merging FITC-labeled nanolipobubbles within a cross-linked hydrogel network. The nanocomposite hydrogel system was characterized by size distribution analysis and zeta potential as well as shears thinning behavior, elastic modulus (G'), viscous loss moduli (G"), TEM, and FTIR. In addition, molecular bio-imaging via Vevo ultrasound and Cell-viZio techniques evaluated the stability and distribution of the nanolipobubbles within the cross-linked hydrogel. FITC-labeled and functionalized nanolipobubbles had particle sizes between 135 and 158nm (PdI = 0.129 and 0.190) and a zeta potential of -34mV. TEM and ultrasound imaging revealed the uniformity and dimensional stability of the functionalized nanolipobubbles pre- and post-embedment into the cross-linked hydrogel. Biomechanical characterization of the hydrogel by shear thinning behavior was governed by the polymer concentration and the cross-linker, glutaraldehyde. Ultrasound analysis and Cell-viZio bio-imaging were highly suitable to visualize the fluorescent image-guided nanolipobubbles and their morphology post-embedment into the hydrogel to form the NanoComposite system. Since the nanocomposite is intended for targeted treatment of neurodegenerative disorders, the distribution of the functionalized nanolipobubbles into PC12 neuronal cells was also ascertained via confocal microscopy. Results demonstrated effective release and localization of the nanolipobubbles within PC12 neuronal cells. The molecular structure of the synthetic surface peptide remained intact for an extended period to ensure potency for targeted delivery from the hydrogel ex vivo. These findings provide further insight into the properties of nanocomposite hydrogels for specialized drug delivery.

Supramolecular Assembly Mediates the Formation of Single-Chain Polymeric Nanoparticles.

A breakthrough innovation in water-based polymeric nanoparticles has enabled significant progress in mimicking the folding of natural proteins by generating novel "single-chain polymeric nanoparticles" (SCPNs) via supramolecular interactions. In this study, a practical approach to the living polymerization of functionalized oligo(ethylene glycol) methacrylate monomers allows the incorporation of self-constituted multiple hydrogen-bonded groups into physically cross-linked polymer networks, which enables the formation of highly functionalized SCPNs in an aqueous environment. The newly developed materials are particularly attractive from a practical point of view since they have a very low critical micellization concentration and uniform particle diameters of ca. 25 nm, making them extremely stable under dilute conditions. Concentration-dependent experiments showed that SCPNs formed at polymer concentrations up to 40 mg/mL with no significant change in morphology observed. Moreover, the formed SCPNs had a very high stability in an aqueous solution containing surfactant, suggesting potential for a wide variety of applications as a promising candidate nanocarrier for bioimaging, controlled release, and drug delivery systems.

A Computational Framework for Bioimaging Simulation.

Using bioimaging technology, biologists have attempted to identify and document analytical interpretations that underlie biological phenomena in biological cells. Theoretical biology aims at distilling those interpretations into knowledge in the mathematical form of biochemical reaction networks and understanding how higher level functions emerge from the combined action of biomolecules. However, there still remain formidable challenges in bridging the gap between bioimaging and mathematical modeling. Generally, measurements using fluorescence microscopy systems are influenced by systematic effects that arise from stochastic nature of biological cells, the imaging apparatus, and optical physics. Such systematic effects are always present in all bioimaging systems and hinder quantitative comparison between the cell model and bioimages. Computational tools for such a comparison are still unavailable. Thus, in this work, we present a computational framework for handling the parameters of the cell models and the optical physics governing bioimaging systems. Simulation using this framework can generate digital images of cell simulation results after accounting for the systematic effects. We then demonstrate that such a framework enables comparison at the level of photon-counting units.

Nanobonding: A key technology for emerging applications in health and environmental sciences

In this paper, surface-activation-based nanobonding technology and its applications are described. This bonding technology allows for the integration of electronic, photonic, fluidic and mechanical components into small form-factor systems for emerging sensing and imaging applications in health and environmental sciences. Here, we describe four different nanobonding techniques that have been used for the integration of various substrates — silicon, gallium arsenide, glass, and gold. We use these substrates to create electronic (silicon), photonic (silicon and gallium arsenide), microelectromechanical (glass and silicon), and fluidic (silicon and glass) components for biosensing and bioimaging systems being developed. Our nanobonding technologies provide void-free, strong, and nanometer scale bonding at room temperature or at low temperatures (<200 °C), and do not require chemicals, adhesives, or high external pressure. The interfaces of the nanobonded materials in ultra-high vacuum and in air correspond to covalent bonds, and hydrogen or hydroxyl bonds, respectively.

Hepatitis B virus preS1-derived lipopeptide functionalized liposomes for targeting of hepatic cells

To enhance the liver-specific delivery, HBVpreS/2-48myr (HBVP), a synthetic HBVpreS-derived lipopeptide endowed with compelling liver tropism, was conjugated to PEGylated liposomes (HBVP-Lip) for hepatic cell-specific delivery. Compared with the non-targeted liposomes, a significantly higher amount of HBVP-Lip were taken up by the primary mice hepatocytes through a receptor-mediated endocytosis mechanism. The endocytosis inhibition assay demonstrated that the endocytosis of HBVP-Lip was mediated mainly by caveolin and clathrin. After systemic administration in mice, HBVP-Lip could be specifically internalized into hepatocytes efficaciously. Furthermore, the hepatoprotective effects of HBVP-Lip loaded with silybin (SLB) on carbon tetrachloride induced acute liver damage were remarkably stronger than the SLB solution and SLB loaded non-targeted liposomes. Preliminary safety results suggested that no acute systemic toxicity or immunotoxicity was observed after intravenous administration with HBVP-Lip. These results indicated that the HBVP-Lip could deliver the payloads to the hepatocytes with high specificity in vitro and in vivo, and raise new possibilities for liver-specific drug delivery systems, gene delivery systems, and bio-imaging systems.