1,554 research outputs found
Unraveling nanoscale alterations in liver cell fenestrations - Morphological studies via optical super-resolution microscopy approaches
The endothelium makes up the innermost cell layer of blood vessels. It consists of a thin layer of simple squamous cells, forming an interface between circulating blood and the surrounding tissue. Endothelial cells of different vascular beds are specialized according to tissue-specific functions. For this project emphasis was placed upon high-resolution methods enabling the study of liver sinusoidal endothelial cells (LSECs) below the diffraction limit of visible light (~200 nm). LSECs have unusual morphology with as much of 20% of their surface covered with cellular fenestrations - holes through the cells of 50-300 nm diameter. These allow bi-directional flow of plasma from the sinusoids to the surrounding hepatocytes, while retaining blood cells in the sinusoidal lumen. Little is known about the function of fenestrations, their regulation, and their role in the transfer of metabolites, viruses, lipoproteins and pharmaceuticals to other cells of the liver.
There are two major challenges with the study of LSEC fenestrations; i) the majority have diameters smaller than the diffraction limit of visible light and; ii) they disappear rapidly in cultured LSEC, and there are no cell line alternatives that express fenestrations. To address the first challenge, the project used classical super resolution imaging technologies such as scanning electron microscopy, and two novel super-resolution optical microscopy modalities: dSTORM (direct stochastic optical reconstruction microscopy) and SIM (structured illumination microscopy) to study the in vitro effects of xanthines, sildenafil and oxidized LDL on LSEC fenestrations. One of the xanthines, theobromine, and sildenafil increased both the frequency and diameter of fenestrations in cultured LSEC. While oxidized LDL caused major disruptions in LSEC fenestration morphology. Finally, to address the second challenge, namely the rapid loss of fenestrations in LSEC, a cryopreservation method for freshly isolated LSEC was developed such that they can be used at researchersâ convenience, rather than directly after isolation from liv
A versatile and customizable low-cost 3D-printed open standard for microscopic imaging
Modern microscopes used for biological imaging often present themselves as black boxes whose precise operating principle remains unknown, and whose optical resolution and price seem to be in inverse proportion to each other. With UC2 (You. See. Too.) we present a low-cost, 3D-printed, open-source, modular microscopy toolbox and demonstrate its versatility by realizing a complete microscope development cycle from concept to experimental phase. The self-contained incubator-enclosed brightfield microscope monitors monocyte to macrophage cell differentiation for seven days at cellular resolution level (e.g. 2âÎŒm). Furthermore, by including very few additional components, the geometry is transferred into a 400 Euro light sheet fluorescence microscope for volumetric observations of a transgenic Zebrafish expressing green fluorescent protein (GFP). With this, we aim to establish an open standard in optics to facilitate interfacing with various complementary platforms. By making the content and comprehensive documentation publicly available, the systems presented here lend themselves to easy and straightforward replications, modifications, and extensions
An Optofluidic Lens Biochip and an x-ray Readable Blood Pressure Microsensor: Versatile Tools for in vitro and in vivo Diagnostics.
Three different microfabricated devices were presented for use in vivo and in vitro diagnostic biomedical applications: an optofluidic-lens biochip, a hand held digital imaging system and an x-ray readable blood pressure sensor for monitoring restenosis.
An optofluidic biochipâtermed the âMicrofluidic-based Oil-Immersion Lensâ (mOIL) biochip were designed, fabricated and test for high-resolution imaging of various biological samples. The biochip consists of an array of high refractive index (n = 1.77) sapphire ball lenses sitting on top of an oil-filled microfluidic network of microchambers. The combination of the high optical quality lenses with the immersion oil results in a numerical aperture (NA) of 1.2 which is comparable to the high NA of oil immersion microscope objectives. The biochip can be used as an add-on-module to a stereoscope to improve the resolution from 10 microns down to 0.7 microns. It also has a scalable field of view (FOV) as the total FOV increases linearly with the number of lenses in the biochip (each lens has ~200 microns FOV).
By combining the mOIL biochip with a CMOS sensor, a LED light source in 3D printed housing, a compact (40 grams, 4cmx4cmx4cm) high resolution (~0.4 microns) hand held imaging system was developed. The applicability of this system was demonstrated by counting red and white blood cells and imaging fluorescently labelled cells. In blood smear samples, blood cells, sickle cells, and malaria-infected cells were easily identified.
To monitor restenosis, an x-ray readable implantable blood pressure sensor was developed. The sensor is based on the use of an x-ray absorbing liquid contained in a microchamber. The microchamber has a flexible membrane that is exposed to blood pressure. When the membrane deflects, the liquid moves into the microfluidic-gauge. The length of the microfluidic-gauge can be measured and consequently the applied pressure exerted on the diaphragm can be calculated. The prototype sensor has dimensions of 1x0.6x10mm and adequate resolution (19mmHg) to detect restenosis in coronary artery stents from a standard chest x-ray. Further improvements of our prototype will open up the possibility of measuring pressure drop in a coronary artery stent in a non-invasively manner.PhDMacromolecular Science and EngineeringUniversity of Michigan, Horace H. Rackham School of Graduate Studieshttp://deepblue.lib.umich.edu/bitstream/2027.42/111384/1/toning_1.pd
Lab-on-a-Chip Fabrication and Application
The necessity of on-site, fast, sensitive, and cheap complex laboratory analysis, associated with the advances in the microfabrication technologies and the microfluidics, made it possible for the creation of the innovative device lab-on-a-chip (LOC), by which we would be able to scale a single or multiple laboratory processes down to a chip format. The present book is dedicated to the LOC devices from two points of view: LOC fabrication and LOC application
Holographic optical trapping Raman micro-spectroscopy for non-invasive measurement and manipulation of live cells
We present a new approach for combining holographic optical tweezers with confocal Raman spectroscopy. Multiple laser foci, generated using a liquid-crystal spatial light modulator, are individually used for both optical trapping and excitation of spontaneous Raman spectroscopy from trapped objects. Raman scattering from each laser focus is spatially filtered using reflective apertures on a digital micro-mirror device, which can be reconfigured with flexible patterns at video rate. We discuss operation of the instrument, and performance and viability considerations for biological measurements. We then demonstrate the capability of the instrument for fast, flexible, and interactive manipulation with molecular measurement of interacting live cell systems
MEMS Technology for Biomedical Imaging Applications
Biomedical imaging is the key technique and process to create informative images of the human body or other organic structures for clinical purposes or medical science. Micro-electro-mechanical systems (MEMS) technology has demonstrated enormous potential in biomedical imaging applications due to its outstanding advantages of, for instance, miniaturization, high speed, higher resolution, and convenience of batch fabrication. There are many advancements and breakthroughs developing in the academic community, and there are a few challenges raised accordingly upon the designs, structures, fabrication, integration, and applications of MEMS for all kinds of biomedical imaging. This Special Issue aims to collate and showcase research papers, short commutations, perspectives, and insightful review articles from esteemed colleagues that demonstrate: (1) original works on the topic of MEMS components or devices based on various kinds of mechanisms for biomedical imaging; and (2) new developments and potentials of applying MEMS technology of any kind in biomedical imaging. The objective of this special session is to provide insightful information regarding the technological advancements for the researchers in the community
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Diagnostic Biosensors for Detection of Blood-Derived Biomarkers
Standard diagnostic tools used from patient samples, specifically from blood draws, require specialized equipment, personnel, and facilities. Conventional techniques can often be very laborious and time consuming due to required sample preparation. The evident delay from sample collection to a patientâs result immensely impacts their outcome. The aims of this research are to design diagnostic biosensors that decrease time-to-results, minimize reagent and sample handling, and incorporate automated simple optical transduction and user interfaces for the detection of blood-derived biomarkers. Specifically, four biosensing detection mechanisms performed on 3 different point-of-care platforms will be discussed.
First is a static loop-mediated isothermal amplification (LAMP) of nucleic acid aqueous droplet on a silicone chip platform immersed in mineral oil. The target-of-interest is a nucleic acid sequence as a biomarker for antibiotic resistant bacteria. The biosensing technique used related changes in interfacial tension (IFT) at the water-oil interface by measuring the change in contact angle (geometrical-effects) over time. Initially the system was characterized as a linear response in relation to concentration of bacteria in a buffer system down to the limit of detection (LOD) of 100 CFU per uL. Subsequently, with the addition of bacterial infected blood sample models, the system became a binary assay (i.e. yes or no) as low as 10 CFU per uL within 10 min of reaction.
Secondly, a two-layered, paper microfluidic chip was utilized to quantify cancer cells from a buffy coat sample matrix by two detection mechanisms: 1) on-chip particle enumeration via smartphone microscope and 2) capillary flow dynamics via smartphone video processing. The assay resulted in a LOD as low as 1 cell per uL for the on-chip imaging aspect of platform and 0.1 cell per uL for the capillary flow analysis within 13 to 22s post application of blood sample.
Lastly, the same concepts previously described in the first platform utilizes changes in IFT due to amplicon presence in an aqueous solution immersed in mineral oil. An emulsion LAMP platform was investigated to determine the relation between angle-dependent light scatter intensity (based off Mie scatter theory) and nucleic acid amplification progression. The phenomenon attributing to changes in light scatter intensities is due to the interfacial changes occurring in the emulsion droplets, where amplicon amount increases the IFT decreases, resulting in smaller diameter emulsions. Changes in light scatter intensity within 3 min of the reaction shows statistical difference in comparison to no target control (NTC) for 10^3 CFU per uL of bacteria dosed into aqueous sample. These four detection mechanisms and three platforms offer but a few alternatives as biosensing methods for blood-derived diagnostic biosensors
CMOS Circuits and Systems for LabâonâaâChip Applications
Complementary metal oxide semiconductor (CMOS) technology allows the functional integration of sensors, signal conditioning, processing circuits and development of fully electronic integrated labâonâaâchip. On the other hand, labâonâaâchip is a technology which changed the traditional way by which biological samples are inspected and tested in laboratories. A labâonâaâchip consists of four main parts: sensing, actuation, readout circuit and microfluidic chamber. Labâonâaâchip gives the promise of many advantages including better and improved performance, reliability, portability and cost reduction. This chapter reviews the currently used labâonâaâchips based on CMOS technology. Also, this chapter presents and discusses the features of the existing CMOS based labâonâaâchips and their applications at the cell level
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