FACS Berg

Flow Cytometry Core Facility - Berg

The FCF-Berg offers „state of the art“ Equipment and Service for Multi-Parameter Analysis and Cell Sorting

  • Basic cell sorts (e.g. Lymphocytes, DCs, cells lines, transfected cells,..)
  • Sorts into 96- or 384-well plates with defined cell numbers (even single cells)
  • Complex sorts under sterile conditions, e.g. use of near UV-laser for stem cell side population of tumor tissue
  • Cell sorting of infectious samples (e.g. primary human samples, virus-transfected cells, bio safety level 2 infected cells)
  • Flow cytometry analysis with up to 16 colors; 561nm-Laser for excitation of RFPs
  • Advice for panel design and analysis of multi-color flow cytometry experiments
  • Seminars on flow cytometry
  • Basic training for users on instruments, software and analysis

The Flow Cytometry Core Facility (FCF) provides the University Clinic Tübingen and external investigators with equipment and support for cell sorting (separation), acquisition, and analysis of flow cytometric data.

In order to use the FCF services, all users have to get registered at FCF-Berg by sending either the filled User Sheet for Sorts or the User Sheet for Analyzers via email to

fcf-berg@med.uni-tuebingen.de

user sheet sorter

user sheet analyzer

 

The next steps are:

 

1. Sorter:

  • appointment for pre-sort consultation
  • Creation of PPMS account for the FCF online booking system

2. Analyzer:

  • Basic Training (mandatory)
  • Creation of PPMS account for the FCF online booking system.

PPMS Homepage

 

Charges & Fees

Basic Training

 

Contact Info

The FCF is equipped with two advanced benchtop Analyzers — BD LSR-Fortessa and BD Canto II, which have up to four lasers (488, 561, 640 and 405 nm excitation wavelengths) and 16 fluorescence channels.

In order to start using the Advanced Analyzers - BD Canto II and BD LSR-Fortessa, users need to:

• Study Virtual Classroom
Basic Training

• Participate in the ”Basic” Flow Cytometry Training

fcf-berg@med.uni-tuebingen.de 

  • Receive date for training via E-Mail from FCF
  • Attend the Training (2x2 hours on the instrument – see fees for price details
  • Get PPMS login/password for the FCF online booking system.
  • Get DIVA login/password to login into DIVA program on BD Canto II or BD LSR-Fortessa

After completion of these steps, researchers have full time access to the analyzers during the daytime hours and can operate the analyzers themselves.

 

Instruments

 

Tipps and Methods

 

Cell sorting is performed by FCF staff on a BD FACS Aria cell sorter equipped with four lasers (488, 561, 640, 355 or 405 nm excitation wavelengths) and 14 fluorescence channel detection. The cell sorter is able to perform high-speed (up to 20,000 events/sec), high-purity (up to 99.9%) sterile sorts (biosafety level 2) into different types of devices (tubes, plates or slides). The cell sorter is operated by FCF staff during the extended business hours.

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

 

Study Virtual Classroom
Basic Training

 

• Participate in the Pre-Sort Consultation

fcf-berg@med.uni-tuebingen.de 

  • Receive confirmation E-Mail from FCF with the date/time specified
  • Attend the consultation. Bring (or have available) all information related to the planned experiment.

To book the Cell Sort, users need to:

  • Login to PPMS System and follow the directions
  • Receive the follow-up confirmation from FCF with the date/time/duration for the Cell Sort originally requested
  • On the date/time of the scheduled Cell Sort come to FCF Sorting Room 581 with controls (compensations and FMO), titrations, and samples with the concentrations suggested at the Pre-Sort Consultation, as well as all buffers, tubes, plates etc.

Notes:

  • 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 29-81979, 29-82745 or 29-87612.
  • Despite the possible delay with the start time, the end time for the sort by default remains unchanged. Please plan accordingly.
  • In some cases the extension of the sorting time is possible (schedule permitted).

To cancel the Cell Sort, users need to:

  • Login to PPMS System and cancel your sort
  • Receive the follow-up confirmation in the PPMS System from FCF

Note: Please refer to Charges and Cancellation Policy

 

To reschedule the existing Cell Sort, users need to:

  • Login to PPMS
  • Cancel your sort
  • Receive the follow-up confirmation in the PPMS System from FCF
  • Book new sort
  • Receive the follow-up confirmation in the PPMS System from FCF with the date/time/duration of the new scheduled Cell Sort

Instruments

Tipps and Methods

The FCF staff members are available for assisted operation of the Analyzers, assisted data analysis, consultation, instrument and software training and troubleshooting from 10:00 am - 5:00 pm weekdays. Please write an E-Mail to FCF-Berg for your specific consultation, training and help need.

 

1. Consultations

  • Experimental Design Consultation
  • Pre-Sort Consultation
  • Pre-Hands-On Consultation

2. Instrument Hands-On Training

  • BD Canto II and BD LSR-Fortessa hands-on Training

3. Instrument Help

  • Help with Instrumental Setup
  • Troubleshooting on the Instrument

4. Software Training (Introduction or Troubleshooting)

  • FlowJo
  • DIVA

Note: In case of emergency or urgent quick questions on the instrument, please do not hesitate to contact FCF-Berg at any time, stop by FCF (Room 580) on weekdays (10:00 am-5:00 pm), or send an E-mail to

fcf-berg@med.uni-tuebingen.de

  during weekends and off hours.

 

Acknowledgements

Please remember to acknowledge the Flow Cytometry Core Facility staff in publications. Co-authorship on publications for FCF staff may be considered appropriate when the staff have provided one or more of the following:

  • Significant intellectual contribution to the design of the published experiments;
  • Substantial practical contribution to the generation, analysis and/or interpretation of experimental data;
  • Indispensable technical support that contributes intellectually or scientifically to the advancement of the work.

FCF staff members who have made an intellectual or technical contribution, but of a limited nature and not justifying authorship as defined above, should be given an appropriate acknowledgement in any resulting publications. This will enable us to demonstrate our involvement in the research of a large number of laboratories at the university, aiding the procurement of future equipment.

Please be sure to send to

fcf-berg@med.uni-tuebingen.de

  the documentation for publications.

SEE YOU SOON IN FCF-Berg!

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