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Concept

Photomicrographie

thumb|Exemple d'image d'un Acarien (Aceria anthocoptes) prise au microscope électronique à balayage La photomicrographie est l'ensemble des techniques photographiques permettant de photographier des sujets à l'aide d'un microscope composé. Le terme désigne aussi l'activité photographique associée à ces techniques. Les rapports de grandissement en photomicrographie se situent en général au-delà du rapport de grandissement 10:1 sur la cible photographique (film ou capteur) (dix fois plus gros sur la cible photographique que la taille réelle). En deçà, et à partir de 1:1, on parle de macrophotographie, technique plus accessible au grand public avec des compléments optiques plus simples à l'appareil photographique. Mais les deux techniques se concurrencent dans le domaine de grandissement de 10:1 à 50:1 car elles ne se distinguent que par l'usage de l'outil, microscope composé, en photomicrographie. La photomicrographie est surtout utilisée en imagerie médicale, pour l'étude des matériaux,

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Morphogenesis of the T4 tail and tail fibers

Petr Leiman

Remarkable progress has been made during the past ten years in elucidating the structure of the bacteriophage T4 tail by a combination of three-dimensional image reconstruction from electron micrographs and X-ray crystallography of the components. Partial and complete structures of nine out of twenty tail structural proteins have been determined by X-ray crystallography and have been fitted into the 3D-reconstituted structure of the "extended" tail. The 3D structure of the "contracted" tail was also determined and interpreted in terms of component proteins. Given the pseudo-atomic tail structures both before and after contraction, it is now possible to understand the gross conformational change of the baseplate in terms of the change in the relative positions of the subunit proteins. These studies have explained how the conformational change of the baseplate and contraction of the tail are related to the tail's host cell recognition and membrane penetration function. On the other hand, the baseplate assembly process has been recently reexamined in detail in a precise system involving recombinant proteins (unlike the earlier studies with phage mutants). These experiments showed that the sequential association of the subunits of the baseplate wedge is based on the induced-fit upon association of each subunit. It was also found that, upon association of gp53 (gene product 53), the penultimate subunit of the wedge, six of the wedge intermediates spontaneously associate to form a baseplate-like structure in the absence of the central hub. Structure determination of the rest of the subunits and intermediate complexes and the assembly of the hub still require further study.

BioMed Central2010

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A Process for Digitizing and Simulating Biologically Realistic Oligocellular Networks Demonstrated for the Neuro-Glio-Vascular Ensemble

Marwan Muhammad Ahmed Abdellah, Marco Agus, Daniya Boges, Corrado Cali, Jay Coggan, Stefan Eilemann, Markus Hadwiger, Renaud Jolivet, Kalpana Kare, Daniel Keller, Pierre Magistretti, Henry Markram, Felix Schürmann

One will not understand the brain without an integrated exploration of structure and function, these attributes being two sides of the same coin: together they form the currency of biological computation. Accordingly, biologically realistic models require the re-creation of the architecture of the cellular components in which biochemical reactions are contained. We describe here a process of reconstructing a functional oligocellular assembly that is responsible for energy supply management in the brain and creating a computational model of the associated biochemical and biophysical processes. The reactions that underwrite thought are both constrained by and take advantage of brain morphologies pertaining to neurons, astrocytes and the blood vessels that deliver oxygen, glucose and other nutrients. Each component of this neuro-glio-vasculature ensemble (NGV) carries-out delegated tasks, as the dynamics of this system provide for each cell-type its own energy requirements while including mechanisms that allow cooperative energy transfers. Our process for recreating the ultrastructure of cellular components and modeling the reactions that describe energy flow uses an amalgam of state-of the-art techniques, including digital reconstructions of electron micrographs, advanced data analysis tools, computational simulations and in silico visualization software. While we demonstrate this process with the NGV, it is equally well adapted to any cellular system for integrating multimodal cellular data in a coherent framework.

2018

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Nanoscopy in nonlinear scanning fluorescence imaging systems

Grégoire Pierre Jean Laporte

In the last 30 years, superresolution in optical microscopy has been a major field of research. During this time, different techniques have been created to break the diffraction limit in order to make observations at a nanometric scale. Given that optical microscopy is non-invasive, those superresolution methods pave the way for a better understanding of biological mechanism at a molecular level. Most of those methods are based on a nonlinear interaction between the excitation light intensity and the sample response (often fluorescent signal). In the same time, nanodiamonds containing fluorescent defects have been proven to be a choice probe for superresolution nanos-copy since they exhibit a strong and stable fluorescent signal even under high light intensities exposure (often required to obtain nonlinear photoresponse). Nanodiamonds containing Nitrogen Vacancy (NV) defects that exhibit a red fluorescent signal had been previously shown to be a viable biomarker for STED superresolved image. First, we demonstrated that green fluorescent nanodia-monds containing Nitrogen-Vacancy-Nitrogen (NVN) defects can be used with a Stimulated Emission Depletion (STED) superreso-lution microscope. Then, we implemented a STED microscope in our lab and compared the properties of NVN and NV centers for STED imaging. We conclude that even if nanodiamonds with NVN defects are less intense, they can be used as a second color nonbleaching biomarker. To illustrate the potential use of green nanodiamonds as bio-compatible probe, we superresolved them internalized into a cell with STED microscopy. Second, we tried to work on one of the main limitation in STED nanoscopy: the lack of information in the axial direction within a single scan. We combined our home made STED microscope with a Double Helix phase mask that modifies the detection point spread function in order to obtain axial localization of the superresolved emitters. We achieved three dimensional localization of nanometric fluorescent emitters but we note that photobleaching was the main limitation of this approach with organic dyes. We discussed different solutions to limit the photobleaching and their feasibility. We also worked on a different superresolution technique that we named Computational Nonlinear Saturated (CNS) microscopy. We showed that with digital post treatment of the acquired data, a nonlinear photoresponse can be harnessed to any scanning microscope equipped with a camera detector to enhance the resolution. We demonstrated that increasing the excitation power and inducing fluorescence saturation, it is possible to break the diffraction limit in a conventional confocal microscope (after data post-treatment). However, with this method, we did not obtain a gain in resolution as high as with other superresolution tech-niques involving fluorescence saturation, such as saturated structured illumination microscopy. To understand the origin of this limitation, we carried out simulation to investigate the performance of CNS microscopy in noisy environments compared with wide field techniques. We propose alternative implementation and quantify the possible resolution gain with simulations. Finally, we demonstrated how a technique, initially created for optical microscopy, can be adapted to lensless endoscopic imaging...

EPFL2016

Chargement

Microscope optique

Le microscope optique ou microscope photonique est un instrument d'optique muni d'un objectif et d'un oculaire qui permet de grossir l'image d'un objet de petites dimensions (ce qui caractérise sa pu

Microscope

Un microscope est un instrument scientifique utilisé pour observer des objets trop petits pour être vus à l'œil nu. La microscopie est la science de l'étude de petits objets et structures à l'aide d'

Microscopie

La microscopie est un ensemble de techniques d' des objets de petites dimensions. Quelle que soit la technique employée, l'appareil utilisé pour rendre possible cette observation est appelé un . De

MSE-352: Introduction to microscopy + Laboratory work

Ce cours d'introduction à la microscopie a pour but de donner un apperçu des différentes techniques d'analyse de la microstructure et de la composition des matériaux, en particulier celles liées aux microscopies électronique et optique. Le cours comprend des cours ainsi que des démonstrations.

MICRO-530: Nanotechnology

This course gives the basics for understanding nanotechnology from an engineer's perspective: physical background, materials aspects and scaling laws, fabrication and imaging of nanoscale devices.

MSE-451: Polymer morphological characterization techniques

Sample preparation and direct observation techniques (optical microscopy, AFM, electron microscopy) and their practical application to the study of morphology and microdeformation in polymers.