English

NovoCyte® Flow Cytometer
NovoCyte flow cytometer system is a high performance benchtop flow cytometer designed for efficient use in clinical laboratories. This budget-friendly
instrument is capable of detecting up to 15 parameters with impressive sensitivity and resolution. The customizable laser and optical configuration of NovoCyte
offers high degree of flexibility while providing complex cell analysis capabilities. The NovoExpress software enables intuitive and easy sample acquisition and
analysis. It allows switching to analysis while still in the process of acquiring other samples. The NovoSampler Pro is an optional high throughput system that
provides versatility in acquiring multiple sample formats such as single tubes, multiple tubes, 24-,48-, 96-well plates in a single, simple setup. Below are listed
the main features: Powerful - up to 15 parameter detection with enhanced sensitivity and resolution; Intuitive - complete acquisition and analysis software,
providing high level of automation and advanced data analysis capability; Customizable - 1 to 3 laser options, exchangeable filters, multiple sampling options
and flexible analysis formats; Volumetric Fluidics System for Increased Accuracy & Easy Maintenance Accurate pressure sensors monitor fluidic status in realtime,
providing warning messages at abnormal high pressures due to possible flow path obstruction; Automated fluidic functions maintain the instrument at an
optimum status, allowing for reliable and accurate data acquisition; Automated SIP (sample injection probe) washing following sample acquisition minimizes
sample carryover and increases capability to detect rare events; High speed data acquisition of up to 35,000 events per second; High Sensitivity and High
Resolution Detection Highly efficient light collection ensures reliable detection of submicron particles and dim signals; Using innovative optical design for light
collection and high quality PMTs, fluorescence signals are efficiently detected with an exceptionally high signal-to-noise ratio, ensuring a high detection
sensitivity for dim markers and small particles; High Fluorescence Signal Resolution Improves Detection Accuracy; Advanced optical and fluidic system
design, premier quality components, and innovative signal processing algorithms collectively ensure accurate extraction of detection parameters with
consistently low variation; Spherotech 8-peak Rainbow Bead.
CytoFLEX Instrument
The advantages of CytoFLEX Research cytometer systems over other instruments include: Up to 21 available configurations to easily adapt to your application
needs with flexibility to obtain exceptionally high levels of information from a single sample. CytoFLEX can acquire up to 15 parameters on a single sample, up
to 13 color fluorescence data channels, plus forward and side scatter. All lower configuration systems have full upgrade capability to higher configurations
enabling future application expansion. CytoFLEX can be easily upgraded. Process samples at sample flow rates providing high sensitivity and adjustable flow
rates, sample sizes as low as 10 μL and a high degree of flexibility in the types of analysis that can be performed. Provides 7 decades of resolution, 24 Bit;
16,777,216 channels resolution and an event collection rate of (30,000 events/sec). Technical specifications enable excellent separation between populations
as well as a large dynamic range of detection to accommodate applications measuring the following: Rare populations, Dim fluorescence, Bright staining index,
or applications that require separation of populations of similar staining. Calibrated sample volume delivery to perform absolute cell counts without a
requirement to use expensive bead products. The full configuration (5/3/5) provides simultaneous measurement of up to 5 fluorescent channels from the
(488nm) Blue laser excitation, 3 fluorescent channels from the (638nm) Red laser, and five CytoFLEX CytExpert Software CytExpert software allows
visualization of data acquisition and data analysis modules and includes a license to CytExpert software at no cost that is designed to collect and analyze flow
cytometry data. CytExpert software allows users to start sample acquisition in 3 mouse clicks or less. Software package is easy to set-up and use. On-line
training module is convenient. Industry standard FCS 3.0 file format supports the use of third party software. Full matrix fluorescence compensation supports
both automatic compensation and manual overwriting for fine tuning. Unique Compensation Library lets users store a repository of compensation spillover
values of dyes in a library to easily determine the correct compensation matrix with virtual multicolor panel and/or with new gain settings. Fully featured analysis
software featuring time saving functions such as linear gain to automatically modify compensation following gain setting changes and auto threshold function
whereby the software automatically sets the threshold based on population scaling in order to easily find target population. CytoFLEX Design The innovative
design of the CytoFLEX flow cytometer offers built¿in system flexibility and superior sensitivity and resolution to get the most of your multicolor analysis.
Compact size 42.5 x 42.5 x 34cm (16.7 x 16.7 x 13.4in) with Tank/Holder 14 x 35.6 x 43.4cm (5.5 x 14.0 x 17.1in) is small enough to allow the unit to be placed
either on a counter or in a laminar flow hood. All instrument fluid containers on board the cytometer without the use of separate fluidics cart reduce space
consumption and simplify operations. Accepts 5mL (12 x 75mm) polystyrene and polypropylene and micro centrifuge 1.5mL and 2mL formats. Minimal weight
requirements 23.4kg (51.6lbs) enabling movement of the instrument if necessary. No special electrical or environmental requirements (i.e. heating, cooling) are
necessary beyond those normally found in a standard laboratory environment. fluorescent channels from the (405nm) Violet laser.

Fonte di Finanziamento: 
Media o grande attrezzatura acquisita/cofinanziata con fondi di Ateneo
anno del bando: 
2018
anno di collaudo: 
2019
Name and acronym of the laboratory or room hosting the Infrastructure: 
Laboratorio di Virologia
Department or host center: 
DIPARTIMENTO DI MEDICINA MOLECOLARE
Laboratory / Infrastructure web page: 
Link al sito del laboratorio
Contatti: 
surnamenamee-mail
Scagnolari
Carolina
carolina.scagnolari@uniroma1.it
Elenco Imprese utenti: 
Elenco altri utenti: 
Ricavi - trasferimenti interni: 
Anno: 
2020
fatture emesse: 
data
05/08/2020
spese manutenzione: 
anno
2020
Description of research activity: 
The mucosal surfaces are densely colonized by commensal bacteria and represents a primary site of interaction between the host and microbes (Freihorst J et 2001). The host immune system has evolved mechanisms to tolerate these commensal organisms while at the same time providing protection for the host from pathogens (Freihorst J et 2001). Therefore, not surprisingly, extensive crosstalk exists between the host and the mucosal surfaces of the respiratory, digestive, and genital tract. In addition to the widely recognized role of commensal bacteria, viruses participate actively in this crosstalk (Almand et al 2017). Indeed, bacteria and viruses often occupy the same niches, however, interest in their potential collaboration in promoting wellness or disease states has only recently gained traction (Moore et al 2018). There are two overarching types of bacterial-virus disease causing interactions: direct interactions that in some way aid the viruses, and indirect interactions aiding bacteria (Almand et al 2017). The virus-promoting direct interactions occur when the virus exploits a bacterial component to facilitate penetration into the host cell. Conversely, indirect interactions result in increased bacterial pathogenesis as a consequence of viral infection. Enteric viruses mainly utilize the direct pathway, while respiratory viruses largely affect bacteria in an indirect fashion. Interestingly mucosal immunity appeared to be strictly controlled in response viral and bacterial pathogens. However, how bacteria can shape the immune response to viral pathogens and how innate and adaptive immune mechanisms drive protective activity against these pathogens remain currently unknown. Indeed, the interaction between commensal microorganisms and host cells in the mucosal surfaces is tightly regulated to maintain homeostasis and to distinguish commensals from pathogens. In this context strategies to defend mucosal surfaces have developed early in evolution and in mammals engage the innate as well as the adaptive arms of the immune system (Durbin RK et al 2013). Host cells express a variety of pattern recognition receptors (PRRs) that aid in the detection of microbes, including Toll-like receptors (TLRs), retinoic acid inducible gene I (RIG-I)-like receptors (RLRs), and nucleotide binding domain and leucine-rich repeat containing receptors (NLRs) (SW Brubaker et al 2015). Upon pathogen recognition by PRRs at the mucosal surfaces, the activation of downstream signalling pathways leads to the production of pro-inflammatory cytokines that are important regulators of the mucosal host response. Among pro-inflammatory cytokines, type I and III interferons (IFN) are mainly produced after viral infection (Yang K et al 2005). While IFNs are universally acknowledged for their antiviral and immunostimulatory functions, there is increasing appreciation of the detrimental effects of inappropriate, excessive, or mistimed type I IFN responses in viral and bacterial infections (Scagnolari et al 2017). Bacterial superinfections following influenza infection are a prominent example of a situation where type I IFNs can misdirect the immune response (Davidson et al 2015). However, less information is available regarding the role of IFN and mucosal cellular immunity during other diseases characterized by viral-bacteria interactions such as HIV, HPV and respiratory diseases. For instance, HIV targets helper T lymphocytes, macrophages, and dendritic cells. By targeting a wide variety of cells within the immune system, its pathogenesis promotes highly complex polymicrobial interactions characterized by bacterial co-infections within the entire human microbiome (Scagnolari et , 2018). It is also known that to prevent over-activation, several feedback mechanisms are in place, which can induce a refractory state to successive stimuli (Nahid et al, 2011; Neagos et al, 2015; de Vos et al, 2009). However, if cells encounter multiple pathogens, this can lead to a lack or delay in the response to secondary infection and breakdown in this immunological process can potentially lead to severe inflammatory disease. The mechanisms employed by the mucosal immune system to maintain this delicate balance are many and varied, and a comprehensive understanding of how they collectively operate would provide novel insight into viral-bacterial interaction in a wide variety of acute or chronic infectious diseases. This analysis can be achieved through flow cytometry technology because it allows the examination of multiple immunonological surface and intracellular markers and can be easily integrate with molecular techniques for determining the transcriptomics, metabolomics, and microbial signature at the mucosal surfaces in different clinical settings.
Keyword iris: 
virologia
microbiologia
immunità innata
malattie infettive
immunità adattativa

© Università degli Studi di Roma "La Sapienza" - Piazzale Aldo Moro 5, 00185 Roma