Equipment

The EVOS Cell Imaging System offers a flexible, easy-to-use platform for fluorescence microscopy, with features that help streamline your imaging workflow while preserving the viability of your cells. Its wide range of objective lenses, multi-colour imaging capabilities, and compatibility with various sample formats make it a valuable tool for researchers looking for reliable and efficient imaging solutions.

Key features

Light source

• LED light: Provides consistent and bright illumination for a variety of fluorophores, offering long-lasting, stable performance.

Objectives

• 4x, 10x, 20x objectives: Flexible magnification options for a range of imaging needs, from broader views to high-resolution detailed imaging.

Stage accommodation

• Compatible with:
  • Regular glass or plastic slides
  • Chamber slides
  • Multi-well tissue culture plates
  • Tissue culture dishes of varying diameters

Channels

• FITC, GFP, Alexa488: For green fluorescence imaging
• Cy3, Rhodamine, Alexa555/568/594: For red fluorescence imaging
• Cy5, APC, Alexa647: For far-red fluorescence imaging
• DAPI: For blue fluorescence (nuclear imaging)
• Bright field: For imaging samples without fluorescence

Key features

3 lasers

• 20mW 488nm direct diode blue laser,
• 20mW 405nm direct diode violet laser, and
• 50mW 561nm Solid State violet laser.

17 parameters

• 1-2 Six fluorescent detectors off 488nm,
• 3-4. Three fluorescent detectors off 405nm,
• 5-8. Six fluorescent parameters off 405nm,
• 9.  Forward scatter, and
• 10. Side scatter.

Acquisition/analysis software

• FACSChorus

The FACSAria-III is an essential tool for researchers who need precise, high-throughput cell sorting with multiple parameters and fluorescence detection. Whether isolating rare cell populations or sorting complex samples, this system provides the flexibility, safety, and performance required for cutting-edge research. With advanced features such as aerosol management, biosafety containment, and multiple sorting options, it is the ideal choice for a wide range of cell biology applications.

Key features

Lasers

The FACSAria-III system is equipped with three lasers for fluorescence detection:

• 488nm blue laser: For a wide range of fluorophores, including FITC, GFP, and Alexa488.
• 633nm HeNe red laser: Provides sensitive detection for red fluorescent proteins, including APC, Alexa647, and Cy5.
• 405nm violet laser: Enables detection of UV-excitable fluorophores like Pacific Blue, DAPI, and BrilliantViolet dyes.

Nozzle sizes

• 70, 85, and 100 microns: These nozzles accommodate most cell types, providing flexibility for sorting a wide range of sample sizes.

Sample sorting

• Sort samples at a single-cell level with precise control over temperature and agitation.
• Sort into various containers, including:
  • 15mL centrifuge tubes
  • 1.0mL microtubes
  • Multi-well plates
  • Microscope slides

Aerosol Management Option (AMO)

• A safety feature that traps aerosolized particles, reducing the risk of contamination and ensuring safe operation in high-risk experiments.

Bio-safety containment

• The FACSAria-III system is housed in a Class-II Type A2 biosafety cabinet, ensuring that your samples remain sterile during the sorting process.

Fluorescent detection channels

The FACSAria-III can detect up to 17 parameters using 15 different fluorochromes. These are distributed across the three lasers and multiple detection channels, allowing for complex multi-parameter analysis. 

Here’s a breakdown of the detection ranges and fluorochromes:

Key features

Sample handling

• Semi-automatic Single Tube Loading with 12 x 75 mm or 1.2 mL tubes
• High Throughput Automated Sample Loader unit for 96 well plates, 96 deep-well plates and 40 tube racks

4 lasers

• 50mW 488nm Solid State blue laser
• 80mW 640nm Solid State red laser
• 100mW 405nm Solid State violet laser
• 50mW 561nm Solid State yellow-green laser

51 parameters

• 1-14: fourteen fluorescent detectors off blue laser
• 15-22: eight fluorescent detectors off red laser
• 23-38:  sixteen fluorescent detectors off violet laser
• 38-48: ten fluorescent detectors off yellow-green laser
• 49: Blue Laser Forward scatter
• 50: Blue Laser Side scatter
• 51: Violet Laser Side scatter

Acquisition/analysis software

• SpectroFlo

Notes

• How to read longpass filter detection range:
  • e.g., 502LP—wavelenge Longer than 502nm will be allowed to Pass through
• How to read bandpass filter detection range:
  • e.g., 530/30 = 530nm ± (30/2)nm = 530nm ± 15nm = from 515nm to 545nm

Key features

Sample handling

• Semi-automatic Single Tube Loading with 12 x 75 mm or 1.2 mL tubes
• High Throughput Automated Sample Loader unit for 96 well plates, 96 deep-well plates and 40 tube racks

5 lasers

• 50mW 488nm Solid State blue laser,
• 80mW 640nm Solid State red laser
• 100mW 405nm Solid State violet laser.
• 50mW 561nm Solid State yellow-green laser
• 20mW 355nm Solid State UV laser

67 parameters

• 1-14: fourteen fluorescent detectors off blue laser
• 15-22: eight fluorescent detectors off red laser
• 23-38:  sixteen fluorescent detectors off violet laser
• 38-48: ten fluorescent detectors off yellow-green laser
• 49-64: sixteen fluorescent detectors off UV laser
• 65: Blue Laser Forward scatter
• 66: Blue Laser Side scatter
• 67: Violet Laser Side scatter

Acquisition/analysis software

• SpectroFlo

The FACSCanto II is ideal for researchers looking for efficiency and precision. Its semi-automatic loading system minimizes operator intervention, saving time and reducing potential errors. With its three lasers and a wide range of fluorescence detection options, it supports a variety of experiments requiring multiple parameters. The system is designed to handle a large volume of samples, making it a valuable tool for both routine and complex analyses.

Key features

Sample handling

• Semi-automatic single tube loading with 12 x 75 mm tubes
• Can automatically load up to 40 tubes without operator intervention

Lasers

• 488nm Solid State blue laser (OmW)
• 633nm HeNe red laser (7mW)
• 405nm Solid State violet laser (OmW)

Detection parameters

• 10 parameters total:
  • 1-4: Four fluorescent detectors off the 488nm laser
  • 5-6: Two fluorescent detectors off the 633nm laser
  • 7-8: Two fluorescent parameters off the 405nm laser
  • 9: Forward scatter
  • 10: Side scatter

Fluorochrome and filter details

The FACSCanto II offers various filter and mirror options for a wide range of fluorochromes. Here are some of the key channels: 

Software

The FACSCanto II uses FACSDiva acquisition and analysis software, which enables easy setup, data collection, and analysis for efficient workflows.

Notes

• Longpass filter detection range: e.g., 502LP—wavelength longer than 502nm will be allowed to pass through.
• Bandpass filter detection range: e.g., 530/30 = 530nm ± (30/2)nm = 530nm ± 15nm = from 515nm to 545nm.

The Fortessa is perfect for researchers needing detailed multicolour analysis, capable of detecting up to 18 fluorescent parameters. Its high-throughput sampler unit speeds up sample acquisition in multi-well plates, while flexible sample handling options save time. Equipped with five lasers (blue, red, violet, yellow-green, and UV), it supports a wide range of fluorochromes and applications, offering versatility and efficiency for complex experiments.

Key features

Sample handling

• Semi-automatic single tube loading with 12 x 75 mm or 1.2 mL tubes
• High throughput sampler unit for multi-well plates, available in standard and high-throughput modes

Lasers

The Fortessa is equipped with five solid-state lasers to enable broad spectrum fluorescence detection:

• 50mW 488nm solid state blue laser
• 40mW 633nm solid state red laser
• 50mW 405nm solid state violet laser
• 50mW 561nm solid state yellow-green laser
• 20mW 355nm solid state UV laser

Detection parameters

• Blue laser (488nm): Detects up to 2 fluorescent parameters
• Red laser (640nm): Detects up to 3 fluorescent parameters
• Violet laser (405nm): Detects up to 6 fluorescent parameters
• Yellow-green laser (561nm): Detects up to 4 fluorescent parameters
• UV laser (355nm): Detects up to 3 fluorescent parameters
• Forward scatter (FSC) and side scatter (SSC) for cell size and granularity analysis

Fluorochrome and filter details

Each laser has specific filters to detect a wide range of fluorochromes: 

Software

The Fortessa uses FACSDiva software for acquisition and analysis, offering an intuitive platform for data collection, visualization, and analysis.

Notes

• Longpass filter detection range: A filter like 502LP allows wavelengths longer than 502nm to pass through.
• Bandpass filter detection range: A filter like 530/30 detects wavelengths within 530nm ± 15nm (515-545nm).

All new users must complete the mandatory instrument training before being granted access to our online booking system. Once the training is completed and budget number provided, the core facility supervisor will add you onto the member list of our online booking system. You will then be able to view the calendar and book the flow cytometry analyzers of your choice at the available time.

Please read to following important guidelines when reserving instrument for your use:

1. Your online booking calendar ID is meant to be used by you ONLY. Please do not share it with others, especially with the untrained users. You should ALWAYS make reservation on the instruments before using the flow cytometers as they are shared by many research groups.
    
2. There are six instruments housed in our facility (FACSCanto-II, LSR-II, FACSAria-III, Cytoflex LX, LSRFortessa, Zeiss Confocal), each instrument has its own calendar. Please select and reserve the correct instrument you have been trained on from the drop-down list on the upper-right corner.
    
3. Please note that you cannot make changes on the event one hour prior to the scheduled time. The instruments, especially the FACSCanto-II, are used heavily by many users, so be considerate to others and reserve only the length of the time you need. ALWAYS cancel your booking in advance if you cannot make it to your appointment. If cancellation is within the same day of your scheduled time, contact me IMMEDIATELY. This is for emergency only and we do not accept booking cancellation after the scheduled booking has passed.
    
4. Although the calendar only shows time from 7am to 6:30pm by default. Instrument reservation is still required for any usage before 7:00 am or after 6:30 pm. You can expand the calendar to see or make bookings before 7:00 am or after 6:30 pm by clicking the small yellow triangle on the top right corner in the Day Grid View.
    
5. You are granted access to reservation of ONLY the instrument you have been trained on (either FACSCanto-II or LSR-II). Additional training is required if you'd like to use the other instrument. Please contact me first to arrange for a training course before reserving the instrument for use.
    
6. Reservation and operation of the FACSAria-III for cell sorting will be done only by the Core Facility Supervisor. Regular users will be able to view but not to book sorting on the calendar. Please check the calendar for available time slot and contact the facility manager by email to book for a cell sorting service.