The Flow Cytometric Cell Analytics and Cell Sorting Core Facility at the LIT offers research groups from the LIT, the University Hospital of Regensburg (UKR), and the University of 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 on flow cytometry-related questions and problems, such as the selection of appropriate fluorochromes, the creation of a staining panel for multicolor analysis and the optimization of gating strategies.

About Assoc. Prof. Hoffmann

FACS Core Facility

Assoc. Prof. Petra Hoffmann heads the FACS Core Facility. She has extensive experience in flow cytometry and provides guidance and support in all aspects of FACS-based cell analysis and sorting such as:

Sample preparation
FACS-based assay systems (e.g. proliferation and cytokine detection)
Multi-parametric cell analysis (e.g. panel design, analysis/gating strategies and dimensionality-reduction approaches)

The facility also employs two operators for the high-speed cell sorters. They are additionally responsible for:

Daily/weekly maintenance work (CS&T, cleaning runs etc.)
Data management (regular backups, archiving sort files)
Help with troubleshooting

Individual flow-cytometry training (basic and advanced) is provided by members of the core facility. In addition, Q&A sessions are organized for user groups with special interests or the need for advanced instrument training. Specialized training is also provided through seminars with external flow-cytometry experts both from academia and within the industry.

Cell Sorting

If you require our services, please refer to the following protocol:

To schedule your sorting appointments, call one of the operators (see ‘Team’) at least one week prior to the intended sorting date.
Appointments can be booked Monday to Friday between 8am and 4pm (other times upon request).
Fill out the FACS Questionnaire (see link below) and bring it with you on the sorting day. Do not forget to provide a signature from your PI.
If you are late for your appointment, please contact your operator (in the lab: +49 (941) 944-5549 or +49 (941) 944-18493 or at the sorter: +49 (941) 944-38191). Delays may result in reduced time allocated for sorting.
Please provide a telephone number for contact on the sorting day.

User information on sample preparation:

The ideal cell concentration for most sorting material is 20 x 10⁶ cells/ml.
The minimum sample volume is 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. For sorting material with biohazard level S2 please contact operators in advance.
Whenever possible provide a flow-cytometric analysis of the intended sorting material (or one as similar as possible) prior to the first sort date.

Data management:

All data acquired during sorting is automatically saved on a local hard drive.
A backup copy will periodically be made on an external hard drive.
All users are also recommended to store their personal data on their own device; files are provided upon request.

Download our questionnaire here!

Cell Analysis

If you require our support, please refer to the following protocol:

Initial registration for the use of cytometers has to be done via the management of the core facility (director, sort operators) or the person responsible for the respective equipment. Upon request, detailed advice can also be provided at this time on the following: specifics regarding cell preparation for acquisition, selection of fluorochromes and setup of staining panels, requirements for automated data acquisition with the HTS, and assistance with data analysis.
Users need to demonstrate adequate basic instrument-handling skills prior to first use of the cytometers on their own. This has to be confirmed by the respective superviser and a correspondent form (provided by the facility management) has to be signed by both and handed in before access is granted.
All subsequent reservations will be made via intranet calendars on a ‘first come, first serve’ basis. Only confirmed users are granted access to the booking calendars.

Terms and conditions

To ensure smooth operations, the following terms and conditions apply:

All bookings are carried out on a ‘first come, first serve’ base.
All users should provide sufficient contact details in the booking calendars.
In case of last minute cancellations or delays subsequent users have to be informed.
Modifications to and interventions with technology and software are carried out exclusively by trained facility staff or after consultation with and approval by the facility’s management.
A logbook is available for each device for each user to note the time of use, name, research group, type of sample material and remarks (such as error messages, unclean workplace, switching off certain lasers, etc.). Each user should sign-off the entry with their initials.
On each cytometer you will find brief instructions listing the most important steps for operating the system; especially those which can lead to system damage or reduction of the life expectancy of important elements (laser, etc.) if performed incorrectly. These instructions must be strictly followed!
The cytometers have to be adequately rinsed and left clean. Liquid waste is to be disposed of appropriately and empty buffer containers have to be refilled. The last user of the day must ensure that the equipment is shut down as instructed.
In case of gross negligence, access to the FACS Core Facility may be blocked and the respective PI will be informed.
Users are liable for damages caused by gross negligence.

Useful links:

Homepage ‘German Society for Cytometry’

Homepage ‘International Society for Advancement of Cytometry’

Selection of the most important publications from the last few years

Bittner S, Ruhland B, Hofmann V, Schmidleithner L, Schambeck K, Pant A, Stüve P, Delacher M, Echtenacher B, Edinger M, Hoffmann P, Rehli M, Gebhard C, Strieder N, Hehlgans T, Feuerer M. Biosensors for inflammation as a strategy to engineer regulatory T cells for cell therapy. Proc Natl Acad Sci U S A. 119(40):e2208436119. 2022.

Meyer M, Jurek B, Alfonso-Prieto M, Ribeiro R, Milenkovic VM, Winter J, Hoffmann P, Wetzel CH, Giorgetti A, Carloni P, and Neumann ID. Structure-function relationships of the disease-linked A218T oxytocin receptor variant. Mol Psychiatry. 27(2):907-917. 2022.

Delacher M, Simon M, Sanderink L, Hotz-Wagenblatt A, Wuttke M, Schambeck K, Schmidleithner L, Bittner S, Pant A, Ritter U, Hehlgans T, Riegel D, Schneider V, Groeber-Becker FK, Eigenberger A, Gebhard C, Strieder N, Fischer A, Rehli M, Hoffmann P, Edinger M, Strowig T, Huehn J, Schmidl C, Werner JM, Prantl L, Brors B, Imbusch CD, Feuerer M. Single-cell chromatin accessibility landscape identifies tissue repair program in human regulatory T cells. Immunity. 54(4):702-720.e17. 2021

Siska PJ, Decking SM, Babl N, Matos C, Bruss C, Singer K, Klitzke J, Schön M, Simeth J, Köstler J, Siegmund H, Ugele I, Paulus M, Dietl A, Kolodova K, Steines L, Freitag K, Peuker A, Schönhammer G, Raithel J, Graf B, Geismann F, Lubnow M, Mack M, Hau P, Bohr C, Burkhardt R, Gessner A, Salzberger B, Wagner R, Hanses F, Hitzenbichler F, Heudobler D, Lüke F, Pukrop T, Herr W, Wolff D, Spang R, Poeck H, Hoffmann P, Jantsch J, Brochhausen C, Lunz D, Rehli M, Kreutz M, Renner K. Metabolic imbalance of T cells in COVID-19 is hallmarked by basigin and mitigated by dexamethasone. J Clin Invest. 131(22):e148225. 2021.

Delacher M, Imbusch C, Hotz-Wagenblatt A, Mallm J, Bauer K, Simon M, Riegel D, Rendeiro A, Bittner S, Sanderink L, Pant A, Schmidleithner L, Braband K, Echtenachter B, Fischer A, Giunchiglia V, Hoffmann P, Edinger M, Bock C, Rehli M, Brors B, Schmidl C, Feuerer M. Precursors for nonlymphoid-tissue Treg cells reside in secondary lymphoid organs and are programmed by the transcription factor BATF. Immunity 52(2):295-312.e11. 2020.

01 | High-speed Cell Sorting

The BD FACSAria™ Fusion High-speed Sorter 1 & 2 offers:

high-speed sorting (max 70.000 events per second)
5 lasers and as many as 18 different colors
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 into 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)

Refer to FACSAria™ Fusion Setup here