Flow Cytometry Equipment

There are three Cytek Aurora spectral analyzers in the flow lab, named Aurora A , Aurora B and Aurora D. All three instruments have five lasers (355 nm, 405 nm, 488 nm, 561 nm, 640 nm), three light scattering channels, 64 fluorescence channels and are equipped with sensor for measuring in real time the sample flow rate. They also have capability of loading samples from standard 96 well plates or from a rack that can hold 48 FACS tube . These cytometers are the most powerful analyzers in our lab and they can be used for detecting over 40 fluorochromes in one sample.

We have two spectral cell sorters in the Flow lab: Aurora CS-1 and Aurora CS-2. They have the same optical configuration as the Aurora analyzers: five lasers (355 nm, 405 nm, 488 nm, 561 nm, 640 nm), three light scattering channels, 64 fluorescence channels with the same set of optical filters. The Aurora CS are versatile sorters that can be used for simple sorts as well as for sorting populations in very complex polychromatic panels. They can be set up with 100 μm or 70 μm nozzles and up to six populations can be sorted at once. Both Aurora CS sorters can deposit cells in multi-well plates.

The BioSorter (Union Biometrica) is a jet-in-air sorter that can be used for separating large particles. Unlike FACSJazz and MoFlo Astrios which continuously generate droplets, the Biosorter relies on the air deflection of a continuous liquid stream into a tray.

It can be set up with two FOCAs (Fluidics and Optics Core Assemblies): 250 μm and 1,000 μm and it is equipped with two lasers and three fluorescence detectors.

The BioSorter can be used for purifying or enriching, large cells (megakaryocytes, adipocytes), cell clusters (pancreatic islets, Drosophila imaginal discs), whole organisms (C. elegans, zebrafish larvae) and various other particles (e.g. beads).

We encourage end-users to learn how to operate the BioSorter and run their-own sorts. Typically one session of approximately 2 hours is sufficient for gaining basic sorting skills on BioSorter.
 

Researchers who prefer to have their samples sorted by the staff should contact us to discuss the project in detail.

Countess links:

• Countess II FL user manual
• Countess II FL Quick Start Guide 

We have three CytoFLEX cytometers in the flow lab, each one equipped with a different number of lasers and with a different optical configuration. Instead of photomultipliers (PMTs) which are commonly used as detectors in most standard flow cytometers, the CytoFLEX devices rely on avalanche photodiode (APD) detectors which are characterized by excellent sensitivity and linearity. The characteristics of APDs make the compensation workflow very efficient. Microparticles as small as 200 nm can be detected on these devices by triggering off the side scatter detector. The CytExpert software used on CytoFLEX is easy to learn and operate. All CytoFLEX cytometers in our lab can be used with 12 x 75mm FACS tubes and also can be set to automatically load samples from standard 96 well plates. The following CytoFLEX devices are available in the flow lab:

• CytoFLEX LX (6 lasers): 21 color detectors
• CytoFLEX S (4 lasers): 15 color detectors
• CytoFLEX S (3 lasers): 15 color detectors.

Evos is a user-friendly inverted imaging system with the following features:

• Fully-automated digital system, fully controlled through software
• Touch-screen monitor
• Long working distance objectives: 1.25x, 4x, 10x, 20x, 40x
• Color camera for bright field and phase-contrast imaging
• Monochrome camera for fluorescence detection
• Up to four colors fluorescence detection. DAPI, GFP, and RFP light cubes are permanently installed. Cy5 light cube can be transferred from the Countess upon request.
• Motorized 3D stage
• Built-in software for scanning slides or plates, time-lapse imaging, Z stacks

Download the Evos FL Auto Quick Start Guide

Evos FL Auto Application Notes:

• Cell Counting
• Tile Stitching
• Time Lapse Imaging
• Z-Stacking
• Phagocytosis

We have two FACSAria sorters, FACSAria Fusion and FACSAria II. Up to four populations can be sorted simultaneously on FACSAria sorters and an Automatic Cell Deposition Unit (ACDU) is available for sorting single or multiple cells in multi-well plates. The FACSAria Fusion has 4 lasers, and 5 color detectors , while FACSAria II has only three lasers and 8 fluorescence detectors. Please note that FACSAria II is not encased in a class II biosafety cabinet and, for this reason, must be used with samples that require containment at BSL-1. FACSAria Fusion has a built in biosafety cabinet and can be used with samples up to BSL-2.

FACSJazz is an electrostatic droplet sorter with blue, yellow-green and red lasers and 6 color detectors. It is encased in a class II biosafety cabinet and it is permanently fitted with 100 μm nozzle. The maximum cell sorting speed is up to 10,000 events/second (or 36 million events/hour) and maximum two populations can be sorted at a time. It has capabilities to sort in multiwell plates and to index sort.

Online reservations must be done at least one day in advance and all reservations for assisted use or full-service sorting have to be approved by the staff in order to become effective.

Users who need to sort cells on FACSJazz at least once a week are encouraged to learn how to use this instrument and take advantage of lower rates. Contact us for more details.

LSR Fortessa

It is equipped with 5 lasers and 18 color detectors. It is operated using the FACSDiva software. In addition to the standard configuration shown above, LSRFortessa can be easily reconfigured to detect YFP and GFP in the same sample.

LSR II

LSR II has four lasers (355, 405, 488 and 640 nm), 15 color detectors and it is operated using FACSDiva, the same software used with LSRFortessa.

LSR links:

• Download FACSDiva Software Quick Reference Guide.