FACS Lab. General Information Page

• Booking Information
• Sample Information
• Instrumentation Available

Booking Information:

Sample Information:

• All samples should be in Falcon #2054 12x75mm tubes- most other tubes won't work on our instruments and will cause you delays and hassles.

• Army tube users please book on the "old" FACScan, and have your
  samples in the tubes before you bring them to the lab., not still in the
  96 well plate.

• Samples should be at a density of 1-2 x10E6 per ml for analysis, higher
  cell concentrations (2-5 x10E6/ml) are preferred for cell sorting, with
  around 4 x 10E6/ml being ideal .

• Unfixed stained samples should be brought to the lab on ice.

• Samples which are virus infected should be fixed in 2% paraformaldehyde in PBS.

• Minimum volume for analysis is 100ul. If cell number is limited, maintain
  the minimum volume.

• Samples may be taken directly from 96 well plates using the Cytek
  but please note that you wish to use it in the booking diary. If you've never
  used the Cytek before please come and have a trial run, or you could end up spraying
  your sample all over the wall!

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Instrumentation Available

• BD-FACStar Plus.

  The Becton Dickinson FACStar Plus is a highly sophisticated flow cytometer. This instrument was purchased in 1992, and is setup with a triple laser bench, fitted with a 488/UV/632 wavelength lasers. This instrument routinely sorts cell populations on the basis of up to six parameters, two light scatter and four fluorescence parameters. Routinely, FITC, and PE (or Propidium iodide) are excited by the primary laser, UV sensitive dyes are excited by the second laser and allophycocyanin (APC) by the the third (632 HeNe) laser. The FACStar Plus is ideal for cell sorting of complex cell populations with high sensitivity and high purity at a relatively rapid sorting speed.
  This instrument is also fitted with an ACDU unit allowing single cell deposition into 96, 24 or Terasaki plates. Temperature control is also available for these different plate arrangements.
  This instrument is also fitted with a MACROSORT option allowing sorting of particles >100uM in diameter.

  
  

  BD-FACScan

  The laboratory has two Becton-Dickinson FACScan flow cytometers. Both of the FACScan's are equipped with three fluorescence detectors, allowing simultaneous analysis of five parameters (two light scatter and three fluorochromes). FITC, PE and one of the red emitting fluorochromes (PI/Tri-colour/Cychrome/PerCP) can be combined. Unlike the above described sorters, the FACScan is engineered with fixed optical, electronic and fluidic components, giving it the flexibility to function as an investigator-operated instrument.
  One FACScan is fitted with a Cytek 96 Well plate sampler, and has a HP 340 computer with HP Pascal operating system.
  The newer FACScan offers pulse width and area measurement, in addition to the usual pulse height, and as such is ideally suited to applications like cell cycle analysis, where use of these "additional" parameters can be informative. This instrument has a Macintosh operating system.
  For advice on using either of the instruments, instrument training etc please see Geoff or Sabine.

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Coulter Multisizer

Click on the small picture of the Coulter Multisizer to see a large (185K) Gif of the instrument. This instrument can be used to not only count cells or other particles, but also give an good indication of the size of the particles. When last tested against standard plastic beads of a known diameter it was accurate to 0.3 uM.

Fisons IAsys Biosensor

Click on the small picture of the Fison's IAsys biosensor to see a large (185K) Gif of the instrument. This instrument can be used in the following applications:

• Concentration determination
• Molecular recognition
• Binding patterns
• Co-operativity
• Mapping of multi-molecular interactions
• Kinetics of association and dissociation

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