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URMC / Research / Flow Cytometry / Library

 

Library

The following reference materials are available for researchers to borrow:

  • Methods in Cell Biology, Volume 75: Cytometry 4th Edition. New Developments. (2004) Darzynkiewick, Roederer and Tanke (eds)
  • Current Protocols in Flow Cytometry
  • Course Notebook - 27th Annual Course in Flow Cytometry
    PROTOCOLS

Frequently Asked Questions:

This page is a work in progress and will be updated regularly.  If you have suggestions please let us know.

Updated as of Nov. 2020 - Here

Connecting to the FCR Database

Connect to the database in order to fill out sort request forms or instrument issues.  For help with sort request forms please click here.

Software Guides

Software guides are available in the "User Docs" folder located on every instrument computer as well as in the FCC_Library (links below) server folder.

Troubleshooting and After Hours

Troubleshooting and after hours guides are available in the "User Docs" folder located on every instrument computer as well as in the FCC_Library (links below) server folder.

Peak 6 Bead Information

The reference bead that is used in the facility is from Spherotech. We use the Rainbow Calibration Particles, 6th peak of 8. The part number for this product is RCP-30-5A-6 and the current lot that we use is AK01.

Connecting to the FCC_Transfer server

Double click the "shortcut to transfer" icon on the desktop of the instrument computer. An explorer window with a folder for each PI will open. NOTE - you will be able to view the contents of the folders but will not be able to read the contents of any of these folders, even your own. These folders are write only from the instruments. As before, drag the folders you want to transfer from the FCS folder to your group folder.

To recover your data on your lab computer:

For PC:

Windows 7:

Click "Start" then in the "Search programs and files" field type:

Medical Center: \\smdnas\FCC_Transfer\PILastName_FirstInitial (ie. bushnell_t)

River Campus: \\smdnas.urmc-sh.rochester.edu\FCC_Transfer\PILastName_FirstInitial (ie. bushnell_t)

 

For Mac:

Click "Go" in Finder, choose "Connect to Server" then type:

Medical Center: smb://smdnas/fcc_transfer/PILastName_FirstInitial (ie. bushnell_t)

River Campus: smb://smdnas.urmc-sh.rochester.edu/fcc_transfer/PILastName_FirstInitial (ie. bushnell_t)


If you are prompted for a username and password use your URMC
network credentials (what you would use for webmail or to log onto a
Windows computer here).

REMEMBER!!

Data will be automatically removed from the transfer partition after two weeks.

Connecting to the FCC_Library server

This is done similarly to the Transfer space. Here we have shared documents that are available to all registered users. This includes past lectures, IDEAS software, and other goodies.

For PC:

Start Menu >In the search menu type:

\\smdnas\fcc_library

For Mac:

Finder >Go > Connect to server

smb://smdnas/fcc_library

If you are prompted for a username and password use your URMC
network credentials (what you would use for webmail or to log onto a
Windows computer here).

Current Instrument Baseline

Available upon request

Grant Information

The URMC Flow Cytometry Resource (FCR), under the scientific direction of Timothy Bushnell, Ph.D. and the technical direction of Matt Cochran, MS., is a full service facility with approximately 1100 sq. feet of newly renovated lab space, employing a staff of 6. The Core currently has traditional analytical tools including two Accuri C6’s (4-colors) and 5 LSR-II’s (one 8 color, one 12-color and three 18-color instruments) from BD Biosciences. We also have a BD 17-color FACSAria-II and an 18-color FACSAria-II for all cell sorting projects. In addition we have several non traditional analytical cytometers. The Amnis ImageStream GenX allows for imaging flow cytometry and the detection of up to 10 fluorescent parameters with cellular localization information. The CyTOF Mass Cytometer, which came online late 2012, uses lanthanide metal mass spectrometry as opposed to traditional fluorescent tags, allowing the analysis of upwards of thirty parameters to be studied simultaneously. The resource also added the Nanosight NS3000 in 2015 for the detection of nano sized particles. The Nanosight is capable of measuring the size distribution and concentration of particles as small as 10 nanometers by using light scatter and Brownian motion. Instrumentation is only part of what we offer. We also offer comprehensive training for staff, consultation for maximizing the effectiveness and cost of your experiments, as well as an environment that fosters success in both existing methods and crossing the boundaries into new frontiers.  All data generated in the FCC is archived onto a 10-terabyte server with daily tape backup.

The FCC has established a series robust QA/QC protocols and procedures that are designed to ensure the instruments are performing optimally. These include standard quality control measurements, as recommended by the instrument manufacturers. Additional detailed instrumentation characterization and QC are performed based on best practices in the field. All QA/QC information is available to investigators and instrumentation characterization data is available on the FCC’s website.

All investigators go through a robust training program to allow them run the analyzers independently, allowing for experienced users to have 24/7 access. All booking is done via an on-line calendar system. All sorting is done by the FCC staff, and is available to investigators from ~10 am till ~8 pm daily.

Recent Publications

  1. Enos, M., Bancos, S., Bushnell, T., and Crispe, I.N. (2008) E2F4 modulates differentiation and gene expression in hematopoietic progenitor cells during commitment to the lymphoid lineage J Immunol 180:3699-707
  2. McGrath, K.E., Kingsley, P.D., Koniski, A..D., Porter, R.L., Bushnell, T.P., and Palis, J.(2007) Enucleation of primitive erythroid cells generates a transient population of "pyrenocytes" in the mammalian fetus. Blood 111:2409-17
  3. Guzman, M.L., Li, X., Corbett, C.A., Rossi, R.M., Bushnell, T., Liesveld J.L., Herbet, J., Young, F., and Jordan C.T. (2007) Rapid and selective death of leukemia stem and progenitor cells induced by the compound 4-benzyl, 2-methyl, 1,2,4-thiadiazolidine, 3,5 dione (TDZD-8). Blood 110:4436-44
  4. Ryan, E.P., Bushnell, T.P., Friedman, A.E., Rahman, I., and Phipps, R.P. (2007) Cyclooxygenase-2 selective inhibition reduces B lymphoma survival by inducing oxidative stress and decreasing intracellular glutathione Cancer Immunol Immunother  57:347-58
  5. Neering, S.J., Bushnell, T., Sozer, S., Ashton, J., Rossi, R.M., Wang, P.-Y., Bell, D.R., Heinrich, D., Bottaro, A, and Jordan C.T. (2007) Leukemia stem cells in a genetically defined murine model of blast crisis CML. Blood 110:2578-85.
  6. Graf, B., Bushnell, T., and Miller , J. (2007) LFA-1-mediated T cell costimulation through increased localization of TCR/class II complexes to the cSMAC and exclusion of CD45 from the immunological synapse. J Immunol., 179:1616-24
  7. Neff-LaFord, H., Teske, S., Bushnell, T.P., and Lawrence, B.P. (2007) Aryl hydrocarbon receptor activation during influenza virus infection unveils a novel pathway of IFN-a production by phagocytic cells J Immunol 179:247-55
  8. Kingsley, PD, Malik J, Emerson RL, Bushnell TP, McGrath KE, Bloedorn LA, Bulger M, and Palis J (2006) “Maturational” globin switching in primary primitive erythroid cells.  Blood 107:1665-1672