BD FACS Canto™ II flow cytometer

Instrument Details

• Make : Becton, Dickinson and Company BD Biosciences
• Model : BD FACS Canto™ II

Specifications :

• Optical platform: Fixed optical assembly
• Beam geometry (all lasers): 9 µm x 65 µm elliptical beam

Emission Optics·

• Collection lens: Optical gel–coupled to flow cell;Numerical aperture (NA) = 1.2
• Fluorescence detection: 6 to 8 photomultiplier tube detectors:
• Wavelength ranges detected from 488-nm laser:750–810 nm (PE-Cy7)
  670–735 nm (PerCP-Cy5.5)
  610–637 nm (PE-Texas Red®, optional)
  564–606 nm (PE)
  515–545 nm (FITC)
  Wavelength ranges detected from 633-nm laser:
  750–810 nm (APC-Cy7)
  701–723 nm (Alexa Fluor® 700, optional)
  650–670 nm (APC)
  Wavelength ranges detected from 405-nm laser:
  502–535 nm (AmCyan)
  425–475 nm (Pacific Blue™)
• Forward scatter detectiont: Photodiode with 488/10 bandpass filter
• Side scatter detection: PMT with 488/10 bandpass filter

Working Principle:The basic principle of flow cytometry is the passage of cells in single file in front of a laser so they can be detected, counted and sorted. Cell components are fluorescently labelled and then excited by the laser to emit light at varying wavelengths. The fluorescence can then be measured to determine the amount and type of cells present in a sample. Up to thousands of particles per second can be analyzed as they pass through the liquid stream. Several detectors are carefully placed around the stream, at the point where the fluid passes through the light beam. One of these detectors is in line with the light beam and is used to measure Forward Scatter or FSC. Another detector is placed perpendicular to the stream and is used to measure Side Scatter (SSC). Since fluorescent labels are used to detect the different cells or components, fluorescent detectors are also in place. The suspended particles or cells, which may range in size from 0.2 to 150μm, pass through the beam of light and scatter the light beams. The fluorescently labelled cell components are excited by the laser and emit light at a longer wavelength than the light source. This is then detected by the detectors. The detectors therefore pick up a combination of scattered and fluorescent light. This data is then analyzed by a computer that is attached to the flow cytometer using special software. The brightness of each detector (one for each fluorescent emission peak) is adjusted for this detection. Using the light measurements, different information can be gathered about the physical and chemical structure of the cells. Generally, FSC can detect the cell volume whereas the SSC reflects the inner complexity of the particle such as its cytoplasmic granule content or nuclear structure.
 

Applications:

• For ploidy analysis in crop varieties (diploid, triploid, tetraploid, hexaploid etc)
• To check the nuclear DNA content of Crop varieties.

Contact Us:

• Director, ICAR-CPRI,Shimla(H.P.)-171001
• Phone: 0177-2625073
• Mobile:+91-9418046415
• Email Id: director.cpri@icar.gov.in