Flow Cytometry

The CytoFlex S is a quick-to-use and easy-to-master benchtop instrument with advanced performance that presents optimal excitation and emission, minimizing light loss and maximizing sensitivity. Relying on fiber optics and band pass filters, its Wavelength Division Multiplier deconstructs and measures multiple wavelengths of light. Light loss due to refraction is minimized as multiple dichroic filters to direct the light path are not needed. High quantum efficiency is achieved by utilizing Avalanche Photodiode Detectors (APD) instead of photomultiplier tubes. To focus light, integrated optics instead of series of lenses and filters are used in the flow cell, ensuring minimal light loss.

Equipped with a 405-nm violet laser, a 488-nm blue laser, 561-nm yellow-green laser and a 640-nm red laser, the CytoFlex S allows the detection of 13 fluorescence and two scatter parameters. Up to 30,000 events per second can be analyzed. APD gain is adjustable by the user, but the 7-decade signal scale and automated data scaling enables analysis even without APD adjustments.

Nanoparticle detection with outstanding sensitivity of particles down to 80 nm is possible with 405 violet side scatter configuration.

Absolute linear gain amplification enables compensation updates with new gain settings. Spillover values can be stored to a compensation library and exported between experiments. Sample acquisition is possible from tubes or a 96 well plate.

CytExpert software is intuitive and easy to learn. It provides automation in quality control, standardization, startup and shutdown. Data is exportable in FCS 3.0 format and the software can be freely installed to personal computers.

Combining accuracy, reproducibility and performance to ease of use, the FACSCanto II is a powerful tool for flow cytometric analyses with multicolor panels and large sample quantities. The fixed-alignment flow cell enables minimal startup times while the optical system provides outstanding signal detection, resolution and sensitivity.

Equipped with a 488-nm blue laser and a 633-nm red laser, the instrument is capable of detecting six compensated fluorescence parameters and two scatter parameters at a rate of 10,000 events per second. PMT voltage is completely adjustable by the user.

The High Throughput Sampler (HTS) supports standard plates of 96 and 384 wells and offers fully automated data acquisition from a well plate as fast as in 15 minutes. --- Automated fluidics startup, shutdown and cleaning procedures simplify workflows and shorten the time consumed at the instrument.

BD FACSDiva™ software controls all the steps of flow cytometric analysis, managing experiment templates, application settings and automated calculations of compensation matrices as well as post-acquisition data analysis. Daily quality control of the cytometer performance is a fundamental feature of the software. Data files can be exported as FCS 3.0 files into third party analysis programs.

FACSAria Fusion offers a very wide repertoire of multicolor analysis possibilities with a state-of-the-art cell sorting capability and bioprotection. The fiber-launched lasers and patented, next-generation quartz cuvette flow cell with fixed alignment ensure experiment reproducibility and data collection efficiency with optimized resolution.

Equipped with four lasers – a 405-nm violet laser, a 488-nm blue laser, 561-nm yellow-green laser and a 640-nm red laser – FACSAria Fusion provides the most versatile analysis options of the core instruments, reaching up to 16 fluorescence and two scatter parameters. Samples can be processed at a maximum rate of 70,000 events/second.

Sorting of up to four populations is possible to different tube formats. Automatic Cell Deposition Unit sorts onto slides and well plates and sorting can be performed at a precision of one cell per well. The facility provides nozzles of 70, 85 and 100 µm for cells of varying size and sensitivity.

Biosafety is taken into consideration by the biosafety cabinet that houses the FACSAria Fusion. It protects the user and the environment as well as the samples from contamination. An independent aerosol management system evacuates any aerosols from the sort chamber. Automation in startup, shutdown and cleaning add user-friendliness and speed to instrument use.

BD FACSDiva™ software in FACSAria Fusion is nearly identical to the software in FACSCanto II, with same features. Having the same software in both cytometers helps users master both instruments. As with the other instruments, FACSAria Fusion data files can be exported as FCS 3.0 files.

For work that requires analysis and sorting of biosafety level (BSL)2, BSL2+ and BSL3 samples in the virus core, the FACSMelody is a great option. Simplified workflow and smart automation shorten considerably the time spent on the sorter, which is highly advantageous when working in a BSL3 environment. The performance of the FACSMelody is, however, not reduced. It has a patented gel-coupled cuvette and fixed laser alignment, guaranteeing optimal resolution of dim cells.

Equipped with a 488-nm blue laser capable of detecting two fluorescence parameters and two scatter parameters, the FACSMelody is suited for simple analysis and sorting of, for example, cells expressing a fluorescent protein. Sorting of up to four populations to tubes is possible.

Automation and ease of use is an essential feature of the FACSMelody. It is ready for sorting in less than 17 minutes, increasing lab efficacy and productivity. BD FACSChorus™ Software monitors and guides every step of instrument usage from startup to quality control, sorting and shutdown, leaving no room for user mistakes. Data files can be exported as FCS 3.0 files.

Reproducibility of results is maximized with spillover values that are automatically updated after quality control runs. New fluorochromes can be added to the software’s list of regents, independent of experimental settings, and spillover can be calculated even if detector gains are changed.

Biosafety in the BSL3 environment is ensured with a HEPA-filtered custom Class II biological safety cabinet and a built-in aerosol management system within the FACSMelody. Sorting with the instrument will be performed by the Virus Facility staff as a service only. The research work must have been authorized by the appropriate national authorities.

The Flow Cytometry Facility offers researchers two FlowJo dongles that enable the use of BD FlowJo software, free of charge. FlowJo is an intuitive and easy to learn analysis software that is compatible with the data files from all flow cytometers in the facility. It features models for simple and advanced analysis in numerical and visual form, with a constantly updating plugin architecture of novel algorithms. Publication quality figures are easy to create with FlowJo. Users can download the software to their personal computers and use it with the dongle at their own office.

By measuring the oxygen consumption rate (OCR) and extracellular acidification rate (ECAR), the Seahorse determines the mitochondrial respiration, glycolysis and ATP production of living cells, providing a systems-level view of cellular metabolic function in cultured and ex vivo cells.

OCR and ECAR are measured at intervals of approximately 5-8 minutes in an extremely small volume of medium above a monolayer of cells. Cellular respiration and glycolysis cause easily measurable changes to the concentrations of oxygen and protons in the medium. The concentrations are measured by solid state sensors for 2-5 minutes and the instrument then calculates the OCR and ECAR. Once a measurement is complete, the small measuring volume is mixed with a larger medium volume, restoring cell values to baseline.

A four-port injection system with automated mixing function allows the label-free detection of cell responses up to four inhibitors, substrates or other compounds in real time. Typically, basal OCR and ECAR are determined first, followed by user-defined addition of drugs at desired intervals and multiple measurements of OCR and ECAR for each condition. Total assay time is usually between 60-90 minutes.

Samples are run on a 24 well format. Instrument use is free, but cartridges and reagents must be provided by the user.