Ricerc@Sapienza

The equipment is characterized by a Cell Imaging tool combined with a Multi-Mode Reader and an incubator-like system for real-time live cell and longer-term kinetic studies. The instrument should work by multiple advanced combined technologies, including fluorescence and high contrast brightfield imaging, monochromator-based ultraviolet and visible (UV-VIS) absorbance, high performance filter-based fluorescence and luminescence. All these functions are controlled and combined by the same software which provides an easy and flexible means for imaging capturing, intensity data detection and output data analysis.
Searching for a possible equipment to gain all these objectives and taking into account that all instruments possess unique and distinctive features, we found that Cytation 1 Cell Imaging Multi-Mode Reader by BioTek Instruments better reflects our requirements. Among other commercially available equipments, multimode detection technology has been employed by different companies for multianalyte testing. For instance, GloMax Explorer Multimode Detection System (Promega Corporation), Varioskan LUX Multimode Microplate Reader (Thermo Fisher Scientific), Mithras LB 940 Multimode Microplate Reader (Berthold Technologies GmbH & Co. KG) and Synergy 2 Multi-Mode Microplate Reader (BioTek Instruments) and Spark multimode microplate reader (Tecan Group Ltd.) possess high performance features as microplate-based readers, based on different technologies (e.g. fluorescence, UV-VIS absorbance, luminescence , fluorescence polarization, etc.) and costs. Nevertheless, along with this tool, additional equipments for imaging and live-cell analysis are required. At the moment, inverted microscope (e.g. Axiovert A1 FL by Zeiss) and microplateincubator systems (es. Ibidi gas and temperature controller) are suitable commercial equipments for our requirements.

Cytation 1 takes advantages of most advanced imaging technologies, among which high intensity LED light sources, filter cubes, hard coated optical fibres and automatic focusing. Also, high-contrast brightfield is the built-in method for transmitted light microscopy which provides significant contrast improvement over standard brightfield, thus allowing powerful and intuitive label-free measurements, such as cell count, proliferation, cell toxicity and confluence.
The fluorescence microscopy module is suitable for both traditional quantitative measurement of fluorescence, and cell imaging, especially during cell-based applications, and more expensive and complex cell imaging, such as 3D spheroids, also taking advantage of Z-stack function, which enables capturing up to 50 slices in a stack and z-projecting the image.
The Multi-Mode Reader works through spectrophotometric methods, based on UV-VIS absorbance, fluorescence and luminescence, suitable for a lot of both biological and nonbiological measurements, including phytochemical quantification of foods and vegetables, pharmaceutical dosage. Also, they can be useful for assaying a wide spectrum of biological activities, for quantifying DNA or RNA, for free-label cell-counting, cell proliferation, apoptosis, necrosis and migration, genotoxicity, enzymatic reactions, kinetic measures, signalling pathways and molecular mechanisms, and for morphology profiling of cell structures and delivery systems.
The incubator-like environmental control (temperature, CO2/O2 control and shaking) ensures optimal conditions for continuous and real-time live cell monitoring and longer-term kinetic analysis.
This instrument is also easy to use without compromising performance, being supported by the advance BioTek's powerful Gen5 software, characterized by a logical and user-friendly interface without requiring extensive user training.
It is designed to easily control and combine all the instrument tools, to easily capture raw data and images (from high magnification, intracellular image analysis, plate reading, ELISA etc.), to analyze data output and transform them into meaningful results.
Furthermore, it allows to use each module separately, by setting the required working protocol, thus making the instrument suitable for both basic and advanced analysis. This software can be also started without connection to instrument thus allowing to perform independently data analysis and elaboration, and to make the instrument available for further users.
Another important advantage of this software is the "Discontinuous kinetics option" to be selected for a long-term kinetic reading with multiple measurements on a plate over any period of time without tying up the reader. With respect to traditional kinetics, this option allows removal of the plate for processing after each read interval, so that other experiments can be conducted with the reader inbetween read intervals. In addition, it also allows flexibility in the read interval: because the plate read is initiated manually the time between reads is not rigidly fixed. When enabled, the Discontinuous Kinetics option uses the internal clock of the PC to provide a time and date stamp to each read event, allowing Gen5 to keep track of the total run time of the entire assay. can be with kinetic curves being generated for each well of each plate automatically by the Gen5 Data Analysis Software.