Italiano

The iBright FL1000 Imaging system is a powerful, easy-to-use western blot and gel documentation system. They provides sensitive, streamlined, multimode image capture. This acquisition image instrument connects to the user through a large capacitive touch screen interface and intelligently designed software to provide stunningly easy western blot and gel imaging. Fluorescent detection offers excellent quantitation and multiplex capability. Chemiluminescent detection delivers the ultimate sensitivity for imaging low abundance proteins. These instruments offers both. Fast Chemiluminescent Detection without film. Capture all your gel and blot images on a single instrument, as DNA, protein, colorimetric or photometric. Dedicated chemiluminescence system with extendable fluorescence capabilities, optional epi or trans-illumination options for blot or gel, adaptable to Spectra Capsules for Epi UV, blue, green, red, NIR, IR fluorescence options on blots.  This imaging system is a compact benchtop instrument which is equipped with five fluorescent channels: three in the visible fluorescence range (RGB) and two in the near infrared (NIR) fluorescence range, which enables compatibility with a wide range of fluorescence dyes. Multiplex up to four channels simultaneously in a single western blot. For example, the iBright FL1000 provides fast, accurate, chemiluminescent detection using an advanced 9.1 MP CCD camera to capture chemiluminescent images with high sensitivity and dynamic range. The incorporated advanced CCD technology results in greater to or equal sensitivity to X-ray film and provides enhanced dynamic range and signal linearity. Capture stained nucleic acid gels (e.g. ethidium bromide, SYBR green, SYBR safe etc.) using the integrated environmentally friendly long-life green LED-based transilluminator, which provides a light source without harmful UV rays or mercury waste from UV transilluminator bulbs. The built-in 12.1 inch LCD touch screen features multi-touch technology that enables pinch-and-zoom, pan-to-move images, and swipe-to-switch between channels and overlays. Acquired images can be easily exported through USB ports, local network drive connection, or Wi-Fi and stored in a secure TFCloud. The TFCloud is web browser-based software tools that are flexible, fast and easy to use. These software tools are compatible with Mac or PC. The Qubit 4 Fluorometer apparatus allows a fast, simple and accurate measurement of RNA quality and the Qubit RNA IQ test provides a quick and simple method to check if an RNA sample has degraded. The assay uses two unique dyes: one that binds to large and intact RNA and another that selectively binds to small and degraded RNA. Together they allow to quickly assess the quality and integrity of an RNA sample. To use, simply add the samples to the RNA IQ working solution, then measure the Qubit fluorometer 4. Save samples and bench space The carefully designed Qubit 4 Fluorometer requires samples of only 1-20 ml. It is intended for use at room temperature and occupies only a small amount of counter space. The Qubit 4 Fluorometer is equipped with advanced optical and data analysis algorithms, is equipped with USB key and cable for data transfer in Excel software and software downloads, as well as a universal power supply, four adapters and CE electromagnetic certification. Calculations and settings are performed automatically by the instrument. Qubit assays for use with the Qubit 4 Fluorometer are all performed using the same general protocol, which uses a simple mix-and-read format with incubation times of only two minutes for DNA and RNA analysis.

Fonte di Finanziamento: 
Media o grande attrezzatura acquisita/cofinanziata con fondi di Ateneo
anno del bando: 
2019
anno di collaudo: 
2020
Nome e acronimo del laboratorio o locale che ospita l'attrezzatura: 
V Piano Edificio PL21
Department or host center: 
Edificio: 
PL021 - Clinica Otorino
Contatti: 
cognomenomee-mail
Greco
Antonio
Numero di utenti per anno: 
10
Elenco Imprese utenti: 
denominazionetipologia
Unità di Biomedicina Traslazionale IBBC CNR
Unità di Ricerca
Elenco altri utenti: 
Utilizzo per ricerca clinica: 
Policlinico Umberto I
Ricavi - trasferimenti interni: 
Anno: 
2020
Importo: 
€35868
fatture emesse: 
data
23/11/2020
spese manutenzione: 
anno
2020
Description of research activity: 
The groups at otolaryngology unit of the Department of Sense Organs will be involved in cancer studies described above to characterise Cancer Stem Cells (CSCs) and Circulating Tumour Cells (CTCs), and identify molecular markers will include in vitro study, and the analysis of tissue and blood samples from patients. In these experiments, the Qubit 4 Fluorometer will be used to quantify proteins and RNA/DNA extracted by cells and tissues, and verify RNA integrity. This last data is particularly crucial when the RNAs have to be amplified and quantified by real-time PCR for detection of low expressed molecules like those characterising cancer stem cells ( topic 1), as well as in the analysed miRNAs in blood of patients with cancer, and subsequent test in culture ( topic 3). The Chemiluminescent detection component of iBright magine acquisition and software will be used for the fast analysis of protein/RNA in Western and Southern blot gels (topic 1-3). The software functions will allow the automatic analyze of up to 4 mini blots or mini gels at a time from a single image, and the accurate selection and identification of lanes and band boundaries for molecular weight estimation, densitometry, or normalization, thus improving the reproducibility and realibility of the results. This instrumental improvement will be of great help in the identification of P75NTR as possible marker in LLSC also allowing the study of p75NTR intracellular proteolytic process, by analysing and quantifying the amount of the different fragments ( full-length; intracytosolic; intranuclear) in relation with cancer stage, and patient response to treatment. Further, the high reproducibility of the data obtained by using the iBright software might also contribute to standardize protocol for liquid biopsy for LLSC patients. The ophtalmology unit will benefit of instrument upgrading for their studies addressed to patient profiling, and to evaluate new therapy for GL and AMD. Beside the routine application of Qubit 4 Fluorometer for protein quantification, both the chemioluminescent and fluorenscent opsions of the iBright improve the sensitivity and the spectrum of molecular analysis. Cheifly: 1) Associated to the measure of neurotrophin content in patient samples by Elisa detection method, the identification of neurotrophin peptides ( pre- and pro-peptide, and mature forms) by their molecular weight (chemioluminecsnce blot mode iBright) allow to correlate Neurotrophin maturation and/or degradation to clinical parameters. 2) Biodistribution analysis of Flagged-rhNGF by application of iBright-FL for detection of near-IR fluorophore 3) Simultaneous detection of diverse molecules on the same gels by using multiple immunofluorescence detection by iBright-FL equipped with diverse RGB filters. 4) Analysis of activation of TrK receptors and/or intracellular signals by contemporaneously detection of phosphorylated and unphosphorylated forms.
Responsabile dell'Attrezzatura: 
antonio.greco@uniroma1.it
Settore ERC: 
LS5_4
Ambiti tecnologici trasversali - Key Enabling Technologies: 
Life-science technologies & biotechnologies
Keyword iris: 
molecular biology
anti-cancer applications
Signal Transducing
Stato dell'attrezzatura: 
In funzione

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