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This paper presents a nanoprobe insertion for nanoinjection applications for biological species based on environmental-scanning electron microscope (E-SEM) nanorobotic manipulation system. Based on the bio-nano manipulation system, we have been proposed a nano-injection method by nanoprobe insertion. The nanoprobe is designed to have a sharp tip to insert the probe into biological specimen. It is...
There are strong implications that Amyloid beta (Aβ) peptide causes neurotoxicity in Alzheimer's disease (AD) through (i) pore formation, (ii) the disruption of ionic channels that could affect calcium homeostasis, and (iii) receptor binding. The actual mechanism(s) remains unclear. In this study, we utilised cell-sized model membranes to observe the stability of lipid vesicles, in real-time, in the...
By using the time lapse of both phase-contrast and fluorescent images of single cells, we investigated the promoter activity. HeLa cells were transfected with EGFP, which encodes EGFP under the control of CMV promoter or cMyc binding site, in the presence of Lipofectamin™ LTX. After transfection with EGFP, phase-contrast and fluorescent images of cells in each well were recorded at intervals of 10min...
We developed Optical pH regulation using functional nanotool impregnated with photo-responsive chemical for selective cell injection of nanotool. The nanotool was modified by fluorescent dye for intracellular measurement. The nanotool was included in the fusogenic liposome. Membrane fusion of the liposome to the cell membrane was used for invasive cell injection of the nanotool. The liposome fuses...
Indirect immunofluorescence (IIF) with HEp-2 cells has been used to detect antinuclear auto-antibodies (ANA) for diagnosing systemic autoimmune diseases. The aim of this study is to develop an automatic scheme to identify the fluorescence pattern of HEp-2 cell in the IIF images. By using the previously proposed two-staged segmentation method, the similarity-based watershed algorithm with marker techniques...
Oxygen is an important parameter with significant effect on cellular development and function. Based on microfluidics, laminar flow and hydrodynamic focusing provide ideal tools for the generation of controlled micro-environments and their application as stimuli to cells. In this paper we demonstrate, for the first time inside a microfluidic device, the generation and characterization of hydrodynamically...
We report on several techniques that have been pursued in our laboratories for packaging complementary metal-oxide semiconductor (CMOS) sensors for use in biological environments, such as cell medium. These techniques are suited for single CMOS die ranging from 1.5 ?? 1.5 mm2 to 3 ?? 3 mm2 in area. The first method consisted of creating high aspect ratio structures from negative-tone photocurable...
In this work, a nano injection system for single cell with small size (diameter: ~5 ??m) is proposed. An environmental scanning electron microscope (E-SEM) nanorobotic manipulation system has been constructed with three units and seven degrees of freedom (DOFs) in total. It can provide a real-time observation with nanoscale resolution during the experiment. Micro pulling and FIB etching are used to...
The potential environmental and health effects of particles at the nanoscale have not been well studied as of yet. Using silica nanoparticles as a model, this study aims to elucidate some of the uncertainties behind nanoparticle -cellular interactions, in order to determine if the ubiquitous presence of these nanoparticles in our environment could be detrimental to public health and the environment...
A micro-optic-fluidic spectrometer is demonstrated for the fluorescence detection of moving particles in a microfluidic chip. The object is excited within an integrated 3D liquid-liquid waveguide and its fluorescence spectrum is real-time recorded with resolution of 10 nm. Three different types of fluorescence beads were tested and distinguished by this spectrometer which holds a promising new concept...
A new probe for scanning near-field optical microscopy (SNOM) that exploits field concentration at a metal tip allows to image single dye molecules at a resolution far beyond the diffraction limit. This probe combines the high resolution of the apertureless SNOM with the single molecule sensitivity of the aperture SNOM. An image of single Cy-3 fluorophores covalently attached to the termini of DNA...
The SK-N-BE(2c) is an I-type neuroblastoma that develops a strong pacing electrical response over time. This response is likely the result of a developing interconnected network, such that the regular pacemaker-like firing of individual cells becomes synchronous over time forming a multi unit spikes signal (MUS). Immuno-fluorescence was used to show the presence of pre and postsynaptic markers, suggesting...
Complex networks underlie the very foundation of neuroscience and the mammalian nervous system, yet high throughput computationally intensive measurements of molecular signaling between large groups of cells and the extraction of statistically significant quantitative information about the underlying network structure is not possible given current techniques. We have developed novel computational...
For real-time imaging of the transmembrane voltage of Jurkat cells, exposed to nanosecond pulsed electric fields, the cells were stained with a voltage sensitive membrane dye (VSD) and illuminated with a 4.8 ns long dye-laser pulse at various time during the electric field pulse. The stained cells were located in a 100 mum stainless steel electrode arrangement mounted at the stage of an inverted microscope...
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