Fluorescence-Activated Cell Sorting Facility Announcements

Featured Announcements

Improvements to the ImagestreamX Imaging Cytometer have finally been scheduled by Amnis Corporation. This instrument will be shipped out for upgrades on June 9th. The analysis computer will remain at Yale but please plan experiments around the June 9th to mid-July frame, when the system will be at the manufacturer.
The upgrade will result in a system similar to their latest model, ImagestreamX MarkII, and will include:
- A new SSC detector that will now allow the simultaneous detection of SSC and PE-Cy7
- A new optical nose pieces with 20x, 40x, 60x objectives for increased resolute
- A new 642 nm laser to replace the 658 nm laser for better separation of APC-Cy7 and Alexa Fluor 647
- Improved fluidics that are extremely stable, minimize foaming and will provide greater consistency and reliability
  • Amistad Stratedigm will be relocated to TAC S633 on June 2nd
  • This price increase is the first since September 2009. It was necessitated by increased service contract, labor and supply costs as well as the need to renew and upgrade some of our aging equipment. Because we lowered our prices on two occasions, our current prices after the increase will be still lower than in 2003.
  • Dr. Ann Haberman, an immunologist and director of In Vivo Imaging Core, now is also the new Director of Cell Sorter Core Facility.
  • The FACS Core has upgraded the 488nm blue laser on the LSRII Green (TAC613) and the 488nm and 640nm lasers on the LSRII (TAC 6).
    Both machines previously had 20mw 488nm lasers; they now are equipped with 50mw 488nm lasers. The LSRII has also been upgraded to a 40mw 640nm laser instead of a 20mw line. Users will notice increased sensitivity and resolution of populations with these new lasers. Users will also notice slight differences in FSC and SSC voltages on these machines. Questions about laser power or filters can be directed to: Ewa.Menet@yale.edu (5-7958) or Geoffrey.Lyon@yale.edu (7-5959) as there are slight difference between each machine. Users can also view the lasers and filters at:http://medicine.yale.edu/labmed/cellsorter/instrumentation/analysis/index.aspx
  • Mario Roederer, Ph.D., Senior Investigator at ImmunoTechnology Section of Vaccine Research Center, NIAID, NIH discovered that one type of "bullet" tube they use for staining leads to large increases in autofluorescence, primarily in the "V450" channel (i.e., excited by near-UV laser, 405 nm, and emitting in the 450 nm range). This is the detector you would use for Pacific Blue, ViViD, or Brilliant Violet 450 dyes. Interestingly, the problem only occurs in the tubes that have been sterilized by gamma-irradiation -- the unsterilized tubes are fine. The tubes are polypropylene, so it's quite possible that this issue may extend to other tubes that are similarly sterilized. The specific tubes come from USA Scientific. The "problem tubes" are catalog #1412-1410; the acceptable tubes are catalog #1412-0400. The sterilization process must be leading to the generation of a UV-excitable dye which leaches into the cells. The longer you incubate cells in the "problem" tubes, the brighter they become. Curiously, this doesn't always happen uniformly -- sometimes half the cells become bright and half do not. Perhaps it requires close proximity of the cell to the plastic. This heterogeneity is insidious -- it looks like you are generating new subsets of cells (rather than having a staining artefact).
  • Stratedigm analyzers: how you can export an experiment or any part of it, including Instrument Settings, and then load it on any of the other Stratedigm machines. Please see the detailed instructions.
  • The FACS Core has upgraded the 488nm blue laser on the LSRII Green (TAC613) and the 488nm and 640nm lasers on the LSRII (TAC 6).
    Both machines previously had 20mw 488nm lasers; they now are equipped with 50mw 488nm lasers. The LSRII has also been upgraded to a 40mw 640nm laser instead of a 20mw line. Users will notice increased sensitivity and resolution of populations with these new lasers. Users will also notice slight differences in FSC and SSC voltages on these machines. Questions about laser power or filters can be directed to: Ewa.Menet@yale.edu (5-7958) or Geoffrey.Lyon@yale.edu (7-5959) as there are slight difference between each machine. Users can also view the lasers and filters at:http://medicine.yale.edu/labmed/cellsorter/instrumentation/analysis/index.aspx