H2020Индивидуална стипендия2019–2021

MitoQuant · Development of Deep-UV Quantitative Microscopy for the Study of Mitochondrial Dysfunction

„Хоризонт 2020“ — Действия „Мария Склодовска-Кюри“

Период: 2019-07-01 → 2021-06-30
Финансиране от ЕС: 202 159 €
Участници: 1
Схема: MSCA-IF-EF-ST

Линиите свързват координатора с партньорите.

Накратко на български

Митохондриите и техните нарушения при болести като диабет и невродегенерация се изучават чрез нов вид микроскопия без използване на флуоресцентни маркери. Това помага за по-точното наблюдение на живи клетки, без да се променя тяхната естествена функция или структура.

Този кратък обзор е генериран от изкуствен интелект

Кратко обяснение, генерирано от езиков модел по текста на CORDIS. Оригиналът е по-долу.

Резултати накратко

Development of Deep-UV Quantitative Microscopy for the Study of Mitochondrial Dysfunction

Mitochondria are potentially the most well-known organelle; they are a fundamental part of the eukaryotic cellular machinery and their dysfunction has been linked to many diseases, ranging from diabetes to neurodegeneration. To gain information on these and other sub-cellular compartments, which contain features well below 200 nm, as well as their interactions, recent research focuses more and more on superresolution microscopy to study sub-cellular and even molecular dynamics. Superresolution microscopy, also called nanoscopy, can resolve details below 100 nm and is on its way to become an indispensable tool in biomedical research. However, almost all optical nanoscopy methods require the sample to be fluorescently labelled to break the diffraction barrier of optical imaging. This is often seen as a serious caveat as photobleaching limits the duration of time-lapse recordings, certain labels require the cell membrane to be damaged, and engineered fusion-proteins can alter the native proteins’ function. A further worry is overexpression, i.e. the cell ends up with far too many proteins of a certain type, which limits the meaningfulness of conclusions from those types of experiments. Therefore, label-free techniques with excellent resolution and specificity are of high interest to alleviate these drawbacks. This is where the MitoQuant project comes into play. Seeing the need for label-free nanoscopy, this project aims to develop a microscopy solution capable of delivering quantitative sub-cellular resolution time-lapse imaging with adequate label-free specificity and contrast that is suitable for organelle imaging. Importantly, and in difference to electron microscopy, this microscope shall be applicable to largely “unprepared” samples and even living cells. Plenty of applications for such a microscope exist. One such example application area, which gives the project its name, is the label-free study of mitochondria and their interaction with other cellular organelles or potentially even individual proteins. The main route to achieving this feat is the exploration of deep ultra-violet (DUV) wavelengths for imaging in combination with quantitative phase microscopy and machine learning. Therefore, the development of a novel instrument to provide adequate resolution and contrast, approaching label-based live-cell superresolution techniques like structured illumination microscopy, is the first main goal of this project. To counter low signals and yet offer highest resolution and specificity, MitoQuant’s second objective is to recover weak signal from mitochondrial by enhancing image quality through novel image processing based on machine learning.

Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз

Цел на проекта

Mitochondria play a vital role in the cellular machinery, hence it is little surprising that their dysfunction has been linked to many diseases, from diabetes to neurodegeneration. However, as many studies on the interplay of organelles and molecular dynamics often employ fluorescence microscopy, a continued worry overshadowing findings and deductions is the possibility that the transfection-induced overexpression of fluorescent proteins skews the obtained results. A recent approach, the gene editor CRISPR-CAS9, which modifies rather than adds DNA sequences, circumvents this issue, but in turn often reduces the available signal levels. To counter low signals and yet offer highest resolution and specificity, MitoQuant aims to image contextual mitochondrial information with label-free superresolution, while simultaneously enhance image quality of specific but sparse fluorescently labelled proteins of interest through recently presented de-noising routines based on machine learning. Therefore, the development of a novel instrument to provide adequate resolution and contrast, matching label-based live-cell superresolution techniques like structured illumination microscopy, is the first main goal of this project. The proposed microscope will work in the deep UV range and employ dedicated optics originally developed for material science to provide high numerical apertures at short wavelengths, thus enabling live-cell imaging in the 100nm range. Concurrently, a neural network will be compiled and trained to enhance signals under low-light conditions and to extract and classify cellular organelles based on their quantitative phase and autofluorescence information. Building on an excellent track record of developing application-tailored microscopes as well as advanced image reconstruction and processing algorithms particularly suited for live-cell superresolution, the researcher strives to start with first live-cell experiments in good time after establishing the technique.

Оригинален текст от CORDIS (на английски).

Участници

UNIVERSITETET I TROMSOE - NORGES ARKTISKE UNIVERSITET · TromsoКоординаторНорвегия