The possibility to perform single cell analysis, the speed of analysis (> 10,000 cells / sec) and the opportunity to analyze up to 20 parameters simultaneously (polychromatic analysis) make flow cytometry a powerful technique to quickly characterize biological multiples profiles (proteins, epitopes, nucleic acids, ion concentration, high content) in a single cell, in a very high number of cells (high throughput). “Deep profiling” of single cell through the diversity of cellular subtypes is complementary to “omics” technologies and is particularly important to understand the complexity of cell biological functions, such as intracellular signal, and mechanisms of differentiation.
The flow cytometry center also makes use of instrumentation that monitors the bioenergetics of live cells, in real time, in 24-well plates. The two main pathways for energy production, mitochondrial respiration and glycolysis, involve cellular oxygen consumption (OCR) and proton efflux (ECAR), respectively. Seahorse XF technology uses sensors to detect extracellular changes in these analytes and measure the rates of cellular respiration, glycolysis and ATP production.
OUR INSTRUMENTS:
- Becton Dickinson LSRFortessa X-20. Benchtop flow cytometer equipped with three lasers (488nm-blue, 640nm-red and 405 nm), with the possibility of being equipped with 5 lasers, for the simultaneous acquisition of 14 fluorescence parameters, in addition to 2 physical parameters. Equipped with automatic acquisition system for reading 96- 384-well plates for high-throughput analysis.
- Becton Dickinson FACSAria Fusion. A cell sorter that delivers high multicolor performance, equipped with five lasers (488nm, 561nm, 640nm, 405 nm e 355 nm), allows the simultaneous analysis of 18 parameters (16 fluorescence and 2 scatter). It is used to separate up to 4 different cell populations simultaneously, based on multidimensional phenotypic and / or functional analyses. Sorted cell subsets (up to 100% purity) are viable and can be grown and / or used for subsequent genomic-functional analyses.
- Seahorse XFe-24 (Agilent). The machine allows to make real time analysis of live cells metabolic activity, by measuring the activity of common metabolic pathways, such as mitochondrial respiration (O2 consumption, OCR) and glycolysis (acidification, ECAR), in order to perform kinetic functional measurements of cell bioenergetics. These analysis are important for either the study of cell response to stimuli, and for the characterization of 2D and 3D cell models like organoids. Moreover, by using specific kits, the Seahorse allows to customize tests for specific metabolic pathways of interest. The Seahorse has a widespread potential of assessing cell functional data, especially when it is used in synergy with other technologies available in our core facility. The wide application range of Seahorse allows to study many bioenergetic parameters in a lot of different physiological and pathological cell types to thoroughly assess critical cell processes such as metabolic activation, proliferation, differentiation, cell death and homeostasis.
Please find below more informations:
Configuration BD-FACSAria Fusion
Configuration BD-LSR-Fortessa X20
Platform Manager: Giulia Finotti (045 8027212)