Search results for "Super-Resolution"

Mutant p53 induces Golgi tubulo-vesiculation driving a prometastatic secretome

2020

TP53 missense mutations leading to the expression of mutant p53 oncoproteins are frequent driver events during tumorigenesis. p53 mutants promote tumor growth, metastasis and chemoresistance by affecting fundamental cellular pathways and functions. Here, we demonstrate that p53 mutants modify structure and function of the Golgi apparatus, culminating in the increased release of a pro-malignant secretome by tumor cells and primary fibroblasts from patients with Li-Fraumeni cancer predisposition syndrome. Mechanistically, interacting with the hypoxia responsive factor HIF1α, mutant p53 induces the expression of miR-30d, which in turn causes tubulo-vesiculation of the Golgi apparatus, leading …

Quantitative super-resolution localization microscopy of DNA in situ using Vybrant® DyeCycle™ Violet fluorescent probe.

2016

Single Molecule Localization Microscopy (SMLM) is a recently emerged optical imaging method that was shown to achieve a resolution in the order of tens of nanometers in intact cells. Novel high resolution imaging methods might be crucial for understanding of how the chromatin, a complex of DNA and proteins, is arranged in the eukaryotic cell nucleus. Such an approach utilizing switching of a fluorescent, DNA-binding dye Vybrant® DyeCycle™ Violet has been previously demonstrated by us (Żurek-Biesiada et al., 2015) [1]. Here we provide quantitative information on the influence of the chemical environment on the behavior of the dye, discuss the variability in the DNA-associated signal density,…

Super-resolution binding activated localization microscopy through reversible change of DNA conformation

2018

ABSTRACT Methods of super-resolving light microscopy (SRM) have found an exponentially growing range of applications in cell biology, including nuclear structure analyses. Recent developments have proven that Single Molecule Localization Microscopy (SMLM), a type of SRM, is particularly useful for enhanced spatial analysis of the cell nucleus due to its highest resolving capability combined with very specific fluorescent labeling. In this commentary we offer a brief review of the latest methodological development in the field of SMLM of chromatin designated DNA Structure Fluctuation Assisted Binding Activated Localization Microscopy (abbreviated as fBALM) as well as its potential future app…

Fluorogenic Tetrazine-Siliconrhodamine Probe for the Labeling of Noncanonical Amino Acid Tagged Proteins

2018

Tetrazine-bearing fluorescent labels enable site-specific tagging of proteins that are genetically manipulated with dienophile modified noncanonical amino acids. The inverse electron demand Diels-Alder reaction between the tetrazine and the dienophile fulfills the criteria of bioorthogonality allowing fluorescent labeling schemes of live cells. Here, we describe the detailed synthetic and labeling protocols of a near infrared emitting siliconrhodamine-tetrazine probe suitable for super-resolution imaging of residue-specifically engineered proteins in mammalian cells.

Advanced Indexing Schema for Imaging Applications: Three-Case Studies

2007

Single molecule localization microscopy of the distribution of chromatin using Hoechst and DAPI fluorescent probes.

2014

Several approaches have been described to fluorescently label and image DNA and chromatin in situ on the single-molecule level. These superresolution microscopy techniques are based on detecting optically isolated, fluorescently tagged anti-histone antibodies, fluorescently labeled DNA precursor analogs, or fluorescent dyes bound to DNA. Presently they suffer from various drawbacks such as low labeling efficiency or interference with DNA structure. In this report, we demonstrate that DNA minor groove binding dyes, such as Hoechst 33258, Hoechst 33342, and DAPI, can be effectively employed in single molecule localization microscopy (SMLM) with high optical and structural resolution. Upon ill…

2019

From the very beginnings of radiotherapy, a crucial question persists with how to target the radiation effectiveness into the tumor while preserving surrounding tissues as undamaged as possible. One promising approach is to selectively pre-sensitize tumor cells by metallic nanoparticles. However, though the “physics” behind nanoparticle-mediated radio-interaction has been well elaborated, practical applications in medicine remain challenging and often disappointing because of limited knowledge on biological mechanisms leading to cell damage enhancement and eventually cell death. In the present study, we analyzed the influence of different nanoparticle materials (platinum (Pt), and gold (Au)…

Diffraction-managed superlensing using metallodielectric heterostructures

2012

We show that subwavelength diffracted wave fields may be managed inside multilayered plasmonic devices to achieve ultra-resolving lensing. For that purpose we first transform both homogeneous waves and a broad band of evanescent waves into propagating Bloch modes by means of a metal/dielectric (MD) superlattice. Beam spreading is subsequently compensated by means of negative refraction in a plasmon-induced anisotropic effective-medium that is cemented behind. A precise design of the superlens doublet may lead to nearly aberration-free images with subwavelength resolution in spite of using optical paths longer than a wavelength. This research was funded by the Spanish Ministry of Economy and…

Desarrollo de productos avanzados para la misión SEOSAT/Ingenio

2016

SEOSAT/Ingenio es la futura misión española de observación de la Tierra en el óptico en alta resolución espacial. Mientras que los productos de imagen a Nivel 1, radiancias geo-referenciadas a nivel de sensor, se encuentran en una fase avanzada de desarrollo existiendo para ello un contrato industrial, los productos de Nivel 2 deben ser desarrollados por los propios usuarios. Este hecho limita el uso de las imágenes a la comunidad científica, restringiendo sus posibles aplicaciones fuera de ésta. Así pues, bajo el marco de un proyecto coordinado y motivados por ofrecer productos de Ingenio/SEOSAT de Nivel 2 a disposición de cualquier usuario, se origina y desarrolla este trabajo. En este ar…

Statistical Performance Analysis of a Fast Super-Resolution Technique Using Noisy Translations.

2014

It is well known that the registration process is a key step for super-resolution reconstruction. In this work, we propose to use a piezoelectric system that is easily adaptable on all microscopes and telescopes for controlling accurately their motion (down to nanometers) and therefore acquiring multiple images of the same scene at different controlled positions. Then a fast super-resolution algorithm \cite{eh01} can be used for efficient super-resolution reconstruction. In this case, the optimal use of $r^2$ images for a resolution enhancement factor $r$ is generally not enough to obtain satisfying results due to the random inaccuracy of the positioning system. Thus we propose to take seve…