Rutgers / Environmental & Occupational Health Sciences Institute (EOHSI)

News

April 2015:

We are launched FluoroFinder program for multicolor immunofluorescence to find fluorochromes and antibodies for your panel design.

Please click here to go to FluoroFinder.

 

Oct. 2012

The new Gallios Flow Cytometer with 10 colors, 3 lasers is available.

 

Jan. 2010:

Core Facility is please to introduce the instrument : BD FACSArray Bioanalyzer.

 

Jan. 2009:

The new state-of-the-art instrument, Beckman Coulter MoFlo XDP Cell Sorter, is avaliable for BL2 cell sorting.

 

 

 

 

 

 

 

 

 

flow_cell

 

 

 

 

 

 

sort stream

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Rutgers

EOHSI

CEED_NIEHS

Beckman Coulter MoFlo XDP Cell Sorter

SP sort
 

The MoFlo allows rapid separation of large numbers of specific cell populations with high purity, recovery and yield. It is an extremely stable platform allowing for analysis at rates up to 100,000 events/second and sort speeds of up to 70,000 events/second depending on the cell type. The instrument is capable of simultaneous 1 - 4 ways sorting. The MoFlo is equipped with 3 solid-state air-cooled lasers allowing for excitation wavelengths of 355, 488 and 633 nm and 11 PMTs making it possible to simultaneously analyze 9 different fluorescent colors. The instrument also has a level 2 biosafety containment cabinet which will permit sorting of transfected human cells. In addition, the MoFlo is equipped with the Cyclone sorting option for single cell cloning.

Excitation
Detectors
Emission
Fluorochromes
488nm
FL1
525nm
FITC, AF488, GFP, YFP, CFSE, Rhodamine123, Vybrant Green
488nm
FL2
575nm
PE, DsRed, Vybrant orange
488nm
FL3
620nm

PE-Texas Red(ECD), PI

488nm
FL4
675nm

PE-Cy5, PerCP, 7AAD

488nm
FL5
755nm

PE-Cy7

355nm
FL6
450nm

AF 350, Hoescht 33342(blue), DAPI, Indo-1

355nm
FL7
630nm

PI, Hoescht 33342 (red)

633nm
FL8
670nm
APC, Alexa Fluor 633, Alexa647
633nm
FL9
740nm
APC-Cy7, AF 633, APC-Alexa750, APC-Alexa770

 

Location:  EOHSI Building, Room 346

 

To make a cell sorting appointment:

The MoFlo XDP cell sorter is only operated by Facility personnel.

To schedule cell sorting, please call the flow lab at 732-445-0211 or email us.

Prior to your cell sorting, please complete the biosafety assessment form and submit it to our office. The form must be received at least 5 days before each scheduled sort as special precautions need to be made for sorting unfixed human cells, or cells with any potentially infectious or biohazard agents.

Please have the following information about your samples ready when you call; cell type, fluorochromes (fluorescent dyes) being used, number of cells in sample, percentage of population in sample to be sorted, what type of receptacle you will sort into (e.g. 5ml polypropylene tube or 96 well plate, etc.).

The PI/Requestor of the sort has the appropriate biosafety approvals from the RU institutional Biosafety Committee (IBC). Material that will require IBC approval includes:

    • Unfixed primary or established human/non-human primate cell lines, and/or;
    • infected/transfected/transformed cell lines (regardless of origin), and/or;
    • Bacteria, viruses, parasites, fungi

    New users need to fill out the User Registration Form and submit the form before/on the first appointment.

     

    Tips for preparation of samples

    • Cell concentration for sorting samples: about 3-5 million cells/ml. It is recommended to bring at least 10 million cells for sorting. You should bring extra sorting buffer in case the sample requires diluting.
    • After typsin treatment for detaching adherent cells in the plate, Soybean Trypsin Inhibitor is recommended to stop trypsinization instead of using serum. You can use ACCUTASE for detaching cells.
    •  

      Basic Sorting Buffer:
      1X Dulbecco's PBS ( Ca, Mg++ Free)
        25 mM HEPES pH 7.0
        1% FBS or BSA (For some cell types, BSA is better than FBS.)

       

    • Cells should be in a minimum volume of 1 ml even if the volume does not give the ideal cell concentration described above.
    • Propidium Iodide (2 g/ml) or DAPI can be added for excluding dead cells during the sorting.
    • Collection Media: You should bring collection media, that range from PBS to enriched tissue culture media with antiboitics or 100% FBS to suit your cells. The sorted cells will be diluted with the sheath fluid.
    • Collection Containers: Sort collection tubes must be polypropylene, 12x75 round bottom test tubes (BD Falcon, Cat. #352063, sterile). The cells can also be collected in 96 well plate, 24 well plate, 384 well plate, or standard slides as well.
    • Cell suspension should be free of cell aggregates and large particles. All samples must be filtered with 40 m nylon cell strainers (BD Falcon Cell Strainer #352340) just before the cell sorting in the sorting lab.
    • Keep cell suspension on ice to prevent aggregation.
    • If you have samples stained with more than one fluorochrome per sample, we recommend you  bring single color stained positive control samples (each fluorochrome individually) and unstained or negative control (isotype) cells for proper compensation.