Search results for " optics"

Noninvasive optical diagnostics of enhanced green fluorescent protein expression in skeletal muscle for comparison of electroporation and sonoporatio…

2015

We highlight the options available for noninvasive optical diagnostics of reporter gene expression in mouse tibialis cranialis muscle. An in vivo multispectral imaging technique combined with fluorescence spectroscopy point measurements has been used for the transcutaneous detection of enhanced green fluorescent protein (EGFP) expression, providing information on location and duration of EGFP expression and allowing quantification of EGFP expression levels. For EGFP coding plasmid (pEGFP-Nuc Vector, 10  μg/50  ml 10  μg/50  ml ) transfection, we used electroporation or ultrasound enhanced microbubble cavitation [sonoporation (SP)]. The transcutaneous EGFP fluorescence in live mice was monit…

FRET-based method for evaluation of the efficiency of reversible and irreversible sonoporation.

2017

It is widely known that not all of the treated cells survive after introduction of exogenous molecules via any physical method. Therefore, it is important to develop methods that would allow simultaneous evaluation of both molecular delivery efficiency and cell viability. This study presents Forster resonance energy transfer (FRET)-based method that allows molecular transfer and cell viability evaluation in a single measurement by employing two common fluorescent dyes, namely, ethidium bromide and trypan blue. The method has been validated using cell sonoporation. The FRET-based method allows the efficiency evaluation of both reversible and irreversible sonoporation in a single experiment. …

2017

Significant progress in nonlinear and ultrafast optics has recently opened new and exciting opportunities for terahertz (THz) science and technology, which require the development of reliable THz sources, detectors, and supporting devices. In this work, we demonstrate the first solid-state technique for the coherent detection of ultra-broadband THz pulses (0.1–10 THz), relying on the electric-field-induced second-harmonic generation in a thin layer of ultraviolet fused silica. The proposed CMOS-compatible devices, which can be realized with standard microfabrication techniques, allow us to perform ultra-broadband detection with a high dynamic range by employing probe laser powers and bias v…

Super-resolved linear fluorescence localization microscopy using photostable fluorophores: A virtual microscopy study

2017

Abstract Current approaches to overcome the conventional limit of the resolution potential of light microscopy (of about 200 nm for visible light), often suffer from non-linear effects, which render the quantification of the image intensities in the reconstructions difficult, and also affect the quantification of the biological structure under investigation. As an attempt to face these difficulties, we discuss a particular method of localization microscopy which is based on photostable fluorescent dyes. The proposed method can potentially be implemented as a fast alternative for quantitative localization microscopy, circumventing the need for the acquisition of thousands of image frames and…

Tunable-frequency three-dimensional structured illumination microscopy with reduced data-acquisition

2018

The performance of a tunable three-dimensional (3D) structured illumination microscope (SIM) system and its ability to provide simultaneously super-resolution (SR) and optical-sectioning (OS) capabilities are investigated. Numerical results show that the performance of our 3D-SIM system is comparable with the one provided by a three-wave interference SIM, while requiring 40% fewer images for the reconstruction and providing frequency tunability in a cost-effective implementation. The performance of the system has been validated experimentally with images from test samples, which were also imaged with a commercial SIM based on incoherent-grid projection for comparison. Restored images from d…

4D (x-y-z-t) imaging of thick biological samples by means of Two-Photon inverted Selective Plane Illumination Microscopy (2PE-iSPIM)

2015

AbstractIn the last decade light sheet fluorescence microscopy techniques, such as selective plane illumination microscopy (SPIM), has become a well established method for developmental biology. However, conventional SPIM architectures hardly permit imaging of certain tissues since the common sample mounting procedure, based on gel embedding, could interfere with the sample morphology. In this work we propose an inverted selective plane microscopy system (iSPIM), based on non-linear excitation, suitable for 3D tissue imaging. First, the iSPIM architecture provides flexibility on the sample mounting, getting rid of the gel-based mounting typical of conventional SPIM, permitting 3D imaging of…

Three-dimensional multiple-particle tracking with nanometric precision over tunable axial ranges

2017

The precise localization of nanometric objects in three dimensions is essential to identify functional diffusion mechanisms in complex systems at the cellular or molecular level. However, most optical methods can achieve high temporal resolution and high localization precision only in two dimensions or over a limited axial (z) range. Here we develop a novel wide-field detection system based on an electrically tunable lens that can track multiple individual nanoscale emitters in three dimensions over a tunable axial range with nanometric localization precision. The optical principle of the technique is based on the simultaneous acquisition of two images with an extended depth of field while …

Femtosecond Structural Dynamics of Proteins

2016

Proteins are the workhorses of living cells, providing essential functions such as structural support, signal transduction, enzymatic catalysis, transport and storage of small ligands. Atomic-resolution structures obtained with conventional X-ray crystallography show proteins essentially as static. In reality, however, proteins move and their motion is crucial for functioning. Although the structure and dynamics of proteins are intimately related, they are not equally well understood. A very large number of protein structures have been determined, but only a few studies have been able to monitor experimentally the dynamics of proteins in real time. In the last two decades, the availability …

Studies towards a directional polychromatic sodium laser guide star

2018

In this work we discuss a mechanism for generation of a coherent source of light from the mesosphere as a new concept of directional laser guide star. In contrast to the near-isotropic spontaneous emission, nonlinear processes in atomic vapors like amplified spontaneous emission can yield highly directional emission in the forward and backward directions. Along with directional emission, excited sodium atoms also radiate at different wavelength creating a polychromatic laser guide star (PLGS). If feasible, a directional PLGS would provide a net gain in the return flux of several orders of magnitude compared to traditional LGS schemes, making possible laser-guided tip/tilt-correction in adap…

View images with unprecedented resolution in integral microscopy

2018

Integral microscopy is a novel technique that allows the simultaneous capture of multiple perspective images of microscopic samples. This feature is achieved at the cost of a significant reduction of the spatial resolution. In fact, it is assumed that in the best cases the resolution is reduced by a factor that is not smaller than ten, what poses a hard drawback to the utility of the technique. However, to the best of our knowledge, this resolution limitation has never been researched rigorously. For this reason, the aim of this paper is to explore the real limitations in resolution of integral microscopy and to obtain optically, without the need of any image-processing algorithm, perspecti…