FCRC JumpStart

The Flow Cytometry Resource Center (FCRC) provides the Rockefeller University investigators with equipment and support for cell sorting (separation), acquisition, and analysis of flow cytometric and image cytometric data.

In order to use the FCRC services, all users have to be registered at FCRC Online Reservation System.   FCRC login/password allows users to use the Flow Cytometry Online Reservation System in order to book time on the Analyzers and Analysis-computers or check time availability of Cell Sorters and ImageStream-X.

You also need to have CFS (RU Central File Storage) login/password in order to transfer data from and to the Analyzers and Analysis-computers, as well as retrieving data from the Cell Sorters. To learn more about the CFS please refer to the RU IT Website at http://it.rockefeller.edu/file-storage.

 

ANALYZERS

The FCRC is equipped with two Advanced benchtop Analyzers — BD LSRIIs, which have up to five lasers (488, 561, 640, 355/or 445 and 405 nm excitation wavelengths) and 16 fluorescence channels. The Center also has one Basic Analyzer - BD Accuri® C6 with two lasers and four fluorescence channels.

In order to start using the Basic Analyzer - BD Accuri® C6, users need to:

    1. Submit the Pre-Hands-On Consultation Form
    2. Receive confirmation email from FCRC with the date/time specified
    3. Attend the consultation. Bring (or have available) all information related to the planned experiments
    1. Submit the BD Accuri® C6 Hands-On Training Form
    2. Receive confirmation email from FCRC with the date/time specified
    3. Book BD Accuri® C6 for the date/time suggested in the FCRC confirmation email
    4. Come to the Hands-on Training with the samples suggested at the Pre-Hands-On Consultation (usually - unstained, all single-color controls and a few samples)

In order to start using the Advanced Analyzers - BD LSRIIs, users need to:

    1. Submit the Flow Cytometry Class Application form for the certain date
    2. Receive confirmation email from FCRC
    3. Attend the Class
    4. Get Diva login/password to login into Diva program on BD LSRIIs
    1. Submit the Pre-Hands-On Consultation Form
    2. Receive confirmation email from FCRC with the date/time specified
    3. Attend the consultation. Bring (or have available) all information related to the planned experiments
    1. Submit the BD LSRII Hands-On Training Form
    2. Receive confirmation email from FCRC with the date/time specified
    3. Book BD LSRII for the date/time suggested in the FCRC confirmation email (LSRII-1 is equipped with 355nm laser, while LSRII-2 is equipped with 445nm laser)
    4. Come to the Hands-on Training with the samples suggested at the Pre-Hands-On Consultation (usually - unstained, all single-color controls and a few samples)

After completion of these steps, researchers have full time (24/7/365) access to the analyzers and can operate the analyzers themselves. In case of essential troubleshooting or help need from FCRC staff, please make sure to fill the Instrument Help Form.

 

SORTERS

Cell sorting is performed by FCRC staff on three BD FACSAria cell sorters equipped with up to six lasers (488, 561, 640, 355, 405, and 445 nm excitation wavelengths) and 18 fluorescence channel detection. All the cell sorters are able to perform high-speed (up to 20,000 events/sec), high-purity (up to 99.9%) sterile sorts into different types of devices (tubes, plates, slides, or custom-made devices). Cell sorters are operated by FCRC staff during the extended business hours. For exact times, please refer to the Cell Sorting and ImageStream-X Schedule.

Notes:

In order to start using the Cell Sorters, users need to:

    1. Submit the Pre-Sort Consultation Form
    2. Receive confirmation email from FCRC with the date/time specified
    3. Attend the consultation. Bring (or have available) all information related to the planned experiment

To book the Cell Sort, users need to:

Notes:

1.  If you cannot make the sorting appointment on time, please make sure to notify about the delay with the start of the sort by calling #7657

2.  Despite the possible delay with the start time, the end time for the sort by default remains unchanged. Please plan accordingly

3.  In some cases the extension of the sorting time is possible (schedule permitted), but needs to be discussed with the FCRC Director

 

 

To cancel the Cell Sort, users need to:

Note: Please refer to the FCRC CHARGES AND CANCELLATION POLICY 

To reschedule the existing Cell Sort, users need to:

Note: In case the sort need to be rescheduled appeared to be within the late cancellation time frame (See FCRC Charges and Fees), two separate forms should be filled:

1. FCRC Service Cancellation Request Form (for original sort time);

2. FCRC SORT Booking Form (for new Sort time)

 

 

IMAGESTREAM-X

The ImageStream-X combines the strength of flow cytometry and fluorescence microscopy in a single platform. It allows for high content assays on rare cells and quantification of biological phenomena with incredible accuracy. The ImageStream-X is able to simultaneously record up to 10-color images with the four-laser excitation (488, 561, 658, and 405 nm wavelengths). Data acquisition on ImageStream-X is performed by FCRC staff during the business hours. For exact times, please refer to the Cell Sorting and ImageStream-X Schedule.

In order to start use the ImageStream-X, users need to:

    1. Submit the Pre-ImageStream Consultation From
    2. Receive confirmation email from FCRC with the date/time specified
    3. Attend the consultation. Bring (or have available) all information related to the planned experiment

To book the ImageStream-X, users need to:

Notes:

1.  If you cannot make the ImageStream-X  appointment on time, please make sure to notify about the delay with the start of the acquisition by calling #7657

2.  Despite the possible delay with the start time, the end time for the acquisition by default remains unchanged. Please plan accordingly

3.  In some cases the extension of the ImageStream-X acquisition time is possible (schedule permitted), but needs to be discussed with the FCRC Director

 

To cancel the ImageStream-X, users need to:

Note: Please refer to the FCRC CHARGES AND CANCELLATION POLICY 

To reschedule the existing ImageStream-X, users need to:

Note: In case the ImageStream need to be rescheduled appeared to be within the late cancellation time frame (See FCRC Charges and Fees), two separate forms should be filled:

1. FCRC Service Cancellation Request Form (for original sort time);

2. FCRC ImageStream Booking Form (for new ImageStream time)

 

To be able to analyze data, users need to:

 

 

STAFF ASSISTANCE

The FCRC staff members are available for assisted operation of the Analyzers, assisted data analysis, consultation, instrument and software training and troubleshooting from 8:00am - 6:00pm weekdays. Please send the request form for your specific consultation, training and help need.

1.    Consultations

2.    Instrument Hands-On Training

3.    Instrument Help

4.   Software Training (Introduction or Troubleshooting)

Note: In case of emergency or urgent quick questions on the instrument, please do not hesitate to contact FCRC at any time, stop by FCRC (DWB205) on the weekdays (8:00am-6:00pm), or send email to FCRC during weekends and off hours.

 

 ACKNOWLEDGEMENTS

 

SEE YOU SOON IN FCRC!