FACS Analytics and Cell Sorting|PD Dr. Petra Hoffmann

The RCI's Central FACS Facility (CFF) offers research groups of the University Hospital Regensburg (UKR) and the University Regensburg (UR) the opportunity to identify and characterize individual cell populations and to isolate them in a targeted manner, thus making them accessible for further downstream analysis. In addition, we offer advice and support for beginners and advanced users with regard to flow cytometric questions and problems, e.g. the selection of appropriate fluorochromes, the creation of a staining panel for multicolor analysis or the optimization of gating strategies.   

The CFF is equipped with a BD FACSAria™ IIu and a BD FACSAria™ Fusion Highspeed Cell Sorter, both for operator-based use. Furthermore, several analytical instruments are available: a BD LSR II™, a BD FACSLyric™, a BD Celesta™ (start: Feb'20) and a BD FACSCalibur™. From March '20 the CFF will also offer access to a 30-parameter FACSymphony™ analyser after appropriate training, including high throughput sample processing. 

On the FACSAria™ IIu cell sorter, a maximum of 9 different fluorochromes (+ FSc/SSc) can be used for targeted FACS-based cell isolation; a maximum of 18 different parameters (+ FSc/SSc) are available on the FACSAria™ Fusion. Both instruments are equipped for 4-way sorting (simultaneous sorting of up to four different populations from one sample); the maximum sorting speed is 70,000 events/sec. Both instruments allow sorting into various vessels, including single cell deposit into microtiter plates, as well as index sorting. Both cell sorters are located in an S2 area and the FACSAria™ Fusion is equipped with an additional safety hood, which is why samples up to safety level S2 (genetic engineering and bio hazard) can be accepted after prior consultation. After each sort, a re-analysis of the isolated population(s) is performed on the device and a sort report is printed. All sort-related FCS files are stored in the CFF and copies are made available to the users.

All further detalis can be found in the respective subfolders.

Alle weiteren Details, Kontaktdaten und Hinweise für Erstnutzer

User guide

New Users

  • To schedule your sort appointments call 944-5549 ( R. Eder or I. Fink) at least ONE WEEK prior to intended sort date
  • Appointments can be booked Monday to Friday between 8 am and 4 pm (other times upon request)
  • If possible provide a flow cytometric analysis of the intended sort material (or one as similar as possible) prior to the first sort date
  • Fill out the FACS Questionnaire (see 'Useful Links') and bring it with you on the sort day (do not forget to provide a signature from your PI)
  • If you are late for your appointment please give us a call (lab: 944-5549 or at the sorter: 944-6644). Delay may result in reduced time for sorting
  • Please provide a telephone number for contact on the sort day 

Cell Sorting - Sample Requirements

  • Ideal cell concentration for most sorting material: 20 x 10e6 cells/ml
  • Minimum sample volume: 0,5 ml
  • Sorting material must be prepared and kept under sterile conditions (exceptions only upon prior request)
  • Cells must be resuspended in sterile PBS (addition of serum/protein possible; please consult operators) and filtered immediately before sorting  (≤ 40µm)
  • PLEASE NOTE: Safety level S1 material (genetic engineering and biohazard) can be sorted without prior notice. Upon special request and ONLY AFTER PRIOR CONSULTATION we might be able to sort material with biohazard level S2.

Data Management

  • all data acquired during sorting is automatically saved on a local hard drive
  • a safety copy will periodically be made on an external hard drive
  • all users are recommended to store their personal data additionally on their own device

Useful Links

Information

Instruments & Locations

BD FACSAria™ IIu

  • high speed sorting(max 70.000 events/s)
  • 3 laser / max 9 different colours & FSc/SSc
  • one- to four-way bulk sorting devices for a variety of tube sizes (50 ml,15 ml,5 ml; Microtubes)
  • BD™ Automated Cell Deposition Unit (ACDU) for sorting to Multiwell plates (24-, 48-, 96-well plates; bulk or single cell deposit)

Location

Central FACS Facility
Building D5/ Ground Floor
Room 3.120
Phone: 944-38191


BD FACSAria™ Fusion

  • high speed sorting (max 70.000 events/s)
  • 5 laser / max 18 different colours & FSc/SSc
  • one- to four-way bulk sorting devices for a variety of tube sizes (50 ml,15 ml,5 ml; Microtubes)
  • BD™ Automated Cell Deposition Unit (ACDU) for sorting to Multiwell plates (24-, 48-, 96-well plates; bulk or single cell deposit)
  • Sort Collection Cooling (Water recirculator for refrigeration or heating)
  • Integrated Biosafety Cabinet (BSC)

Location

Central FACS Facility
Building D5/ Ground Floor
Room 3.120
Phone: 944-38191


BD FACSCalibur™

  • 2 laser / max. 4 different colours & FSc/SSc
  • Data acquisition via BD CellQuest Pro

Location

Central FACS Facility
Building D5/ 1st Floor
Room 4.120
Phone: 944-38192


BD LSR II

  • 3 laser/ max. 13 different colours & FSc/SSc
  • Data acquisition via BD FACS Diva Software

Location

Central FACS Facility
Building D5/ 1st Floor
Room 4.120
Phone: 944-38192


BD FACSCelesta™

  • 3 laser/ max. 12 different colours & FSc/SSc
  • Data acquisition via BD FACS Diva Software

Location

Central FACS Facility
Building D5/ 1st Floor
Room 4.120
Phone: 944-38192


BD FACSLyric™

  • 3 laser/ max. 10 different colours & FSc/SSc
  • Data acquisition via BD FACSuiteTM software

Location

Central FACS Facility
Building D5/ 1st Floor
Room 4.120
Phone: 944-38192

Facility Staff

PD Dr. Petra Hoffmann
Head
Phone: +49 941 944-38492
petra.hoffmann(at)ukr.de

Rüdiger Eder
Chief Operator
Phone: 944-5549
ruediger.eder(at)ukr.de

Jaqueline Dirmeier
Operator on parental leave
Phone: +49 941 944-18493
jaqueline.dirmeier(at)ukr.de

Irina Fink
Operator
Phone: +49 941 944-18493
irina.fink(at)ukr.de

Dr. Michael Delacher
responsible for LSR
Phone: +49 941 944-38125
michael.delacher@ukr.de

Important Contact Information

To schedule an appointment:

  • Rüdiger Eder                 phone: +49 941 944-5549
  • Irina Fink                       phone: +49 941 944-18493
  • Jaqueline Dirmeier       phone: +49 941 944-18493
  • Dr. Michael Delacher    phone:+49 941 944-38125
    (LSR) 

To reach operators during sorting

  • Aria                                  phone: +49 941 944-38191

For consulting on

  • sample preparation
  • multicolour panels (composition/gating/analysis)
  • FACS-based assay systems:

Petra Hoffmann                        phone: +49 941 944-38492

News

The CFF has moved and is now located in the Research Building D5, Rms 3.120 (Sorter) and 4.120  (Analysis)

Maps