Note di applicazione

Visualization of microglia-neuron junctions with super-resolution and confocal microscopy

luglio 2020

Microglia are the main immune cells in the brain, and play roles in brain homeostasis and neurological diseases. However, the fundamental mechanisms underlying microglia-neuron communication remain unclear. Dr. Csaba Cserép, Dr. Balázs Pósfai and colleagues, (Laboratory of Neuroimmunology led by Dr. Ádám Dénes, Institute of Experimental Medicine) identified an interaction site between neuronal cell bodies and microglial processes in the mouse brain and studied the function of microglia (C. Cserép and B. Pósfai et al., Science 10.1126/science.aax6752 (2020)). In this Application Note, we introduce how the structure of neuron-microglia junctions was revealed on a nano scale resolution using the confocal and super-resolution microscopes.

Imaging sodium transients in neurons and astrocytes

maggio 2020

Mapping brain cell types with CARTANA in situ sequencing on the Nikon Ti2-E microscope

gennaio 2020

In situ sequencing (ISS) kits offered by CARTANA allow scientists to detect transcripts from hundreds of genes directly inside tissue sections with unprecedented throughput and single-cell resolution. The Nikon Ti2-E inverted microscope enables this application by combining its Perfect Focus System 4 focus locking technology and an automated stage with large image scan capabilities. This note demonstrates ISS for mapping transcriptional cell types within the spatial context of mouse brain sections.

Nikon NIS-Elements Software: utilizing deep learning to denoise confocal data

gennaio 2020

Noise is a fundamental component of confocal images, a result of discreet digital sampling of continuously emitting photons from samples. The contribution of noise to image quality (signal-to-noise ratio) increases as the signal decreases as a square-root function. Using a trained neural network, we use artificial intelligence to remove the shot noise component from confocal image data, allowing an increase in image quality and the ability to acquire dimmer samples at faster rates. NIS-Elements software’s deploys this trained network for live or post-acquisition processing.

Ti2: Increasing data collection and fidelity by maximizing field of view

settembre 2019

With the release of the Nikon Ti2 inverted microscope, the world’s first 25mm FOV became available. Now, Nikon has taken advantage of this improvement by building the largest FOV point scanner in the world, the A1 HD25®.

A1R HD25: the latest in resonant scanning technology allows new live-cell imaging approaches

febbraio 2019

Capturing the dynamics of living systems requires high acquisition rates. Large samples, such as whole model organisms, additionally require a large field of view. The Nikon A1R HD25 confocal system provides both, combining Nikon’s improved HD high speed resonant scanner with an unprecedented 25 mm field of view. The performance of this system is evaluated in zebrafish embryos.

Increasing Data Collection and Fidelity by Maximizing Confocal Field of View

novembre 2018

For years, the field of view (FOV) of confocal systems has been limited by the FOV of the microscope they are attached to. With the release of the Nikon Ti2 inverted microscope, the world’s first 25-mm FOV became available. Now, Nikon has taken advantage of this improvement by building the largest FOV point scanner in the world, the A1 HD25. This Application Note focuses on the impact of this technology on simple, everyday experiments.

Structured Illumination Microscopy (SIM) Imaging Comparison with Confocal

settembre 2018

The super-resolution microscopy technique structured illumination microscopy (SIM) imaging of dendritic spines along the dendrite has not been previously performed in fixed tissues, mainly due to deterioration of the stripe pattern of the excitation laser induced by light scattering and optical aberrations.

Reflectance imaging for visualization of unlabeled structures using Nikon A1 and N-SIM

gennaio 2018

Reflectance imaging allows the user to form an intensity image from light backscattered by the sample. Highly reflective markers, including a variety of nanoparticles, allows for imaging with very high signal-to-noise and virtually free of photobleaching, ideal for both confocal and structured illumination microscopies.

Hardware Triggering: Maximizing Speed and Efficiency for Live Cell Imaging

dicembre 2017

Live cell imaging experiments now require higher speeds and more data throughput than ever before. Nikon Instruments has robust tools that enable hardware triggering of imaging devices in microscopy via direct signaling between hardware. This minimizes delays, synchronizes devices, and reduces the exposure of specimens to light. This Application Note explains how Nikon’s NIS-Elements hardwaretriggering workflow operates, and details its benefits for common time-lapse acquisition routines.

Application of Patterned Illumination Using a DMD for Optogenetic Control of Signaling

novembre 2017

Digital micromirror devices (DMDs) are powerful tools for photostimulation applications, including photoconversion and optogenetic manipulation, owing to their robust ability to produce novel illumination patterns with high spatiotemporal resolution. In this Application Note we showcase recent work describing how DMD technology integrated into a Nikon system can be applied toward light-gated optogenetic control of intracellular signaling.

Robotic Microscopy with the Nikon Ti2 for High-Content Analysis Applications

dicembre 2016

Robotic Microscopy—a combination of high-content screening methods—enables multivariate experimental approaches with large cell populations and member-level sensitivity. Here we explore how the new Nikon Ti2 line of inverted research microscopes is uniquely suited to Robotic Microscopy applications, focusing on work utilizing induced pluripotent stem cells (iPSCs) as disease models in drug screening.

N-STORM with DNA-PAINT for Reliable Multicolor & 3D Single Molecule Localization Imaging

novembre 2016

N-SIM for Quantitative Ultra-Structural Analyses of the Nuclear Lamina

ottobre 2016

Super-resolution Structured Illumination Microscopy (SIM), available from Nikon via the N-SIM and N-SIM E systems, allows for the observation of details inaccessible to traditional microscopes, such as confocal and widefield. In this application note we see how the N-SIM system enables quantitative multi-color evaluation of the distribution of different nuclear lamin proteins and the structures they form.

Quantitative Cluster Analysis Applications for N-STORM

ottobre 2016

Nikon’s N-STORM super-resolution microscope is applied to mapping the distribution of nucleosomes on chromatin fibers, shining a light on the mechanisms of chromatin folding, gene expression, and pluripotency. Multicolor STORM is used to explore how nucleosome distribution differs between differentiated and pluripotent stem cells and correlates with the distribution of other important factors in replication, such as RNA Polymerase II.

Nikon’s Large-Format Multiphoton System for Intravital Imaging

dicembre 2015

Nikon’s A1R MP multiphoton confocal microscope system provides a choice platform for deep tissue imaging. Researchers at the Allen Institute for Brain Science have applied the A1R MP towards fast dual-color intravital imaging of the mouse brain. The open architecture instrument design allows for the addition of custom stages for large samples, external photostimulation devices, and more – perfect for constantly evolving needs.

Quantitative Analysis Tools and Correlative Imaging Applications for N-STORM

novembre 2015

Stochastic Optical Reconstruction Microscopy (STORM) has had a significant impact on our understanding of nano-scale biological processes. In this application note, we see how Nikon’s N-STORM system, combined with quantitative analysis tools, enables single molecule level studies of cannabinoid signaling in the brain. Correlative 3D-STORM, confocal microscopy, and patch-clamp electrophysiology is also explored.

Ricerca sulle cellule staminali con Nikon

maggio 2014

This flyer provides a quick visual orientation of example Nikon products and how they support various research aspects of stem cell biology and regenerative medicine. This includes detection of differentiation markers, routine cell culture checking, time lapse imaging, and more.

Sviluppo di un bioprocesso integrato per la produzione di cellule NK-92 per l'immunoterapia

gennaio 2014

L’uso dei linfociti anti-cancro NK-92 si è dimostrato promettente in immunoterapia. Questa nota applicativa esplora un flusso di lavoro completo per la produzione di cellule NK-92 di grado clinico, utilizzando la BioStation CT di Nikon per monitorare il fenotipo e la potenza dei NK-92.

Test di citotossicità cinetica linfocitaria antitumorale (collaborazione con CCRM)

gennaio 2013

In questa nota applicativa esploriamo l'uso della piattaforma di microscopia di screening ad alto contenuto Nikon BioStation completamente robotizzata per quantificare il tasso di mortalità di un modello di linea cellulare di leucemia umana co-incubata con i linfociti antitumorali NK-92. Questo test integra l'imaging automatizzato della fluorescenza di Calcein-AM per eseguire l’analisi di cellule vive/morte.

Metodo non invasivo per il conteggio delle cellule staminali pluripotenti umane con imaging di cellule vive

gennaio 2012

Presentiamo un metodo per il conteggio delle cellule staminali pluripotenti vive utilizzando la colorazione fluorescente nucleare SYTO24 e l'imaging a contrasto di fase sul microscopio di screening ad alto contenuto della Nikon BioStation CT. I risultati mostrano che l'area di copertura delle colonie dedotta dalle immagini a contrasto di fase può essere utilizzata di per se come stima accurata del solo conteggio delle cellule staminali, correlando con le misurazioni basate sull'emocitometro.

Schema delle applicazioni delle cellule staminali

gennaio 2011

The Nikon BioStation CT cell culture observation system and CL-Quant software provide a powerful platform for stem cell research applications, including automated induced pluripotent stem (iPS) cell colony identification, proliferation analysis, and neural differentiation analysis. Real experimental data is provided for each of these use cases.

Test di screening per le risposte PSC ai substrati

gennaio 2011

An automated imaging-based screening assay for assessing stem cell pluripotency is described. Researchers from Nikon and the Centre for Commercialization of Regenerative Medicine use a Nikon BioStation CT with CL-Quant software to perform stem cell pluripotency maintenance assays in experiments testing feeder cell-free culture vessel substrates. Such commercially available substrates provide a number of process standardization benefits.

Analisi automatica di cellule formanti colonie (CFC) ematopoietiche

gennaio 2011

Il sistema di screening per cellule vive ad alto contenuto Nikon BioStation CT viene utilizzato in un'analisi automatizzata per il conteggio e l'identificazione di cellule formanti colonie ematopoietiche. Questo test consente l'identificazione di diversi tipi di colonie, aiutando a distinguere tra i diversi lineage.

Generazione di una curva di crescita per cellule iPS in coltura senza feeder con analisi di immagini non invasiva

gennaio 2011

In questa applicazione viene dimostrato l’uso del sistema Nikon BioStation CT per l'osservazione con time-lapse prolungato di colture di cellule staminali senza strato di feeder, insieme al rilevamento automatizzato delle colonie di cellule staminali. Le curve di crescita, sia per le cellule differenziate che non, sono state generate in un periodo di tempo di circa 1 settimana.

Creazione di un algoritmo per il rilevamento automatico di cellule iPS/non-iPS in condizioni di coltura mediante analisi di immagine non invasiva

gennaio 2011

In questa nota applicativa viene descritto un metodo automatizzato ad alto rendimento per osservare e classificare le colonie di cellule staminali con un algoritmo standardizzato utilizzando la Nikon BioStation CT. L'analisi automatizzata dell'immagine discrimina le colonie di cellule staminali dalle colonie di cellule non staminali utilizzando l'imaging a contrasto di fase e la successiva analisi delle caratteristiche morfologiche distintive.

Sistema automatizzato, non invasivo di cultura e valutazione per cellule iPS in differenziamento neuronale

gennaio 2011

Questa nota applicativa descrive un flusso di lavoro integrato per la coltura, l'osservazione, la valutazione e la differenziazione di cellule staminali pluripotenti indotte (iPS) in neuroni, introducendo metodi di processamento di immagine per localizzare le rosette neurali e lo sviluppo di neuriti - due importanti checkpoint morfologici nella differenziazione neuronale. L'imaging automatizzato per questo test viene eseguito utilizzando una Nikon BioStation CT.

Risposta delle cellule staminali embrionali a substrati disponibili in commercio per la manutenzione delle cellule staminali

gennaio 2011

In collaborazione con il Centro per la Commercializzazione della Medicina Rigenerativa (CCRM), sviluppiamo un saggio che utilizza il microscopio e il sistema di incubazione di screening ad alto contenuto di Nikon BioStation CT per confrontare le prestazioni di 8 substrati comuni per la crescita e il mantenimento delle cellule staminali. L'imaging a fluorescenza di Oct4 viene utilizzato come marker per la pluripotenza.

Analisi della Sopravvivenza di Neuroni Umani derivati in vitro utilizzando la nuova tecnologia di imaging di Time-lapse prolungato

gennaio 2011

In questa nota applicativa dimostriamo l’uso della Nikon BioStation CT per l'imaging prolungato di cellule vive su motoneuroni derivati da cellule staminali, tracciandone caratteristiche chiave, come la dimensione del corpo cellulare e la lunghezza dei neuriti, prima, durante e dopo il trattamento con diversi fattori di stress. Questo approccio fornisce un metodo standardizzato per gli studi di drug discovery in modelli di malattia neurodegenerativa.