Flow Cytometry Core

BD FACSCanto II™

The Becton-Dickinson FACSCanto II™ cytometer is an easy to operate 3-laser (8 colour) analytical flow cytometer.

• 405nm 30mW -2 detectors (450/50BP, 525/50BP)
• 488nm 20mW – 4 detectors (530/30BP, 585/42BP, 670LP, 780/60BP)
• 633nm 17mW – 2 detectors (660/20BP, 780/60BP)

The patented optical design features fiber optics and a gel coupled cuvette flow cell. Fluorescent signals are collected using highly efficient octagon/trigon collection detector arrays that use serial light reflections to guide signals to their target detectors, resulting in highly efficient light collection and providing maximum signal retention at the detector level. This BD serial reflective design further enhances instrument sensitivity by collecting the dimmest emission signals first, moving from the longest wavelengths (typically PE-Cy™7) to the shortest (FITC).to maximize signal detection, increasing sensitivity and resolution. The sample injection tube features a rinse function that automatically washes down the probe between tubes, to reduce sample cross contamination.

The cytometer runs BD FACSDiva acquisition software.

FACScan

The Becton-Dickinson FACScan is an easy to operate single laser analytical flow cytometer. Excitation is via a 488 nm laser line. In addition to the usual scatter parameters, 3 fluorescence colors may be detected for each cell – fixed filters are green (530/30), orange (585/42), and red (650 LP). The instrument has a pulse-processing option for doublet discrimination.

1. Neither cytometer nor computer will not turn on.
   Make sure that UPS under bench is turned on.
2. Computer will not connect to cytometer.
   Restart computer.
3. Cytometer status is "NOT READY".
   Allow to warm up for 5 minutes.
   Check sheath is full and waste is empty.
   If status window says that waste is full and you know it is empty, rap on waste tank to dislodge sensor float.
   Check that tubing leading to tanks is not pinched or obstructed.
4. Cytometer status remains at "STANDBY".
   Fluidics control not turned to "RUN"
   Tube is cracked.
   Check that pressure is ON (vent valve UP).
   Check that sheath tank is actually pressurized – it should not be moveable.
   Ensure that sheath tank cap is screwed down tightly.
   Stuck fluidics – turn to STANDBY and back to RUN.
5. Status READY, but sample will not run.
   No cells in sample.
   Clumpy sample – filter it.
   Blocked SIP or flow cell – Remove sample, then DRAIN/FILL 3 times.
   Waste not venting – loosen waste tank cap. Report problem.
6. Sample runs for a few seconds then stops.
   Clumpy sample – filter it.
   Blocked SIP or flow cell – Remove sample, then DRAIN/FILL 3 times.
   Waste not venting – loosen waste tank cap. Report problem.
7. Sample still will not run.
   Air bubbles in sheath filter – purge filter.
   SIP blocked – Set fluidics to "HIGH". Run 100% bleach for 30 sec. followed by at least 2 changes of DI water for 1 minute.

LSRII

The Becton-Dickinson LSR II is a 5-laser 15-color analytical flow cytometer. The cytometer runs BD FACSDiva acquisition software.

Configuration

Please contact staff if your fluorochome cannot be accommodated by the standard filters shown; we have other filters in our collection.

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FACS Vantage SE DiVa

The Becton-Dickinson FACS Vantage SE DiVa is a high-speed multi-color multi-laser cell sorter.

Excitation Sources

• Coherent CUBE 405 solid state laser: 100 mW at 407 nm
• Coherent Sapphire 488nm solid state laser: 200 mW at 488 nm
• Coherent Sapphire 568nm solid state laser: 200 mW at 568 nm

Becton-Dickinson (BD) DiVa™ digital electronics enable generation of pulse height, area, and width for detected parameters. Regular or large cells may be sorted. DiVa™ electronics allows sorting of up to 4 populations simultaneously, or cells may be sorted into multiwell plates (up to 96w plate format).

BD InFlux™

The Becton-Dickinson Influx™ cytometer is a state-of-the-art, 13 parameter 5-laser high-speed jet-in-air cell sorter.

Excitation Sources

• 355nm 60mW – 2 detectors (460/50BP, 670/30BP)
• 405nm 100mW -2 detectors (460/50BP, 520/35BP)
• 488nm 200mW – 2 detectors (530/40BP, 692/40BP)
• 561nm 150mW – 3 detectors (593/40BP, 670/30BP, 750LP)
• 640nm 120mW – 2 detectors (670/30BP, 750LP)

To achieve very sensitive, low noise electronics, the BD Influx system uses analog electronics in the first stage of signal amplification. The output of the analog peak and hold circuitry is digitized using 16-bit ADCs and placed on a patented parallel processing digital bus. The bus handles real-time compensation as well as other digitized signals, including area and width of up to 8 additional pulses.

The Influx has several nozzle sizes available to accommodate a variety of particle sizes and can sort 6 populations simultaneously (for suitable sample types). Cells may be sorted into a variety of tube types or multiwell plates (e.g. 384 well).

Additionally, the BD Influx™ is equipped with an aerosol management option; air is drawn away from the operator through a HEPA filter, reducing the amount of aerosols in the sort chamber for an added level of safety.

Quick Links

• Fluorochrome Table
• BD Spectra Analyzer
• Biolegend Spectra Analyzer
• Molecular Probes Spectra Viewer
• Biocompare – Antibody search tool
• Flowjo – Tutorials

Education & Reference

• Practical Flow Cytometry– 4th Edition (Howard Shapiro)
• Flow Cytometry: A Basic Introduction (Wiki) (Mike Ormerod)
• BD Introduction to Flow Cytometry (on-line course)
• Selecting Reagents for Multicolor Flow Cytometry (Holden Maecker, Joe Trotter)
• Interpreting flow cytometry data: a guide for the perplexed (Herzenberg et al)
• Cytokine Encyclopedia
• Common phenotypic markers (Mouse and Human)
• Maturation markers reference
• Pathways database