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Niesner lab

We develop technologies for intravital imaging and live cell imaging to achieve new insights into chronic inflammatory processes

Introduction
Members
Cooperation partners
Third party funding projects
Selected Publications

Biophysical Analytics

Recently, we developed and refined an endogenous fluorescence method for in vivo microscopy to monitor the NAD(P)H-dependent enzymatic activity in cells. This method allows us to distinguish between various classes of enzymes based on the spatially-resolved fluorescence lifetime of the coenzymes NADH and NADPH. Using this technique, we were able to define the concept of “oxidative stress memory” in the context of chronic neuroinflammation and neurodegeneration, both in mouse models and in patients. We found that astrocytes and microglia within the central nervous system (CNS) of diseased mice show a partially irreversible phenotypic shift towards oxidative stress generation, which is also present in monocytes of MS patients. Due to the persistence of plasma cells in the CNS in the late disease phases, we assumed and found that antibodies maintain this phenotypic shift of astrocytes in human brain slices treated with a NMDA-R antibody. This project is a collaboration with Dr. H. Radbruch and Prof. Dr. F. Heppner (Neuropathology, Charité), Prof. Dr. F. Paul (Neuroimmunology, Charité) and Prof. Dr. A.E. Hauser (DRFZ). Since we expect a similar phenotypic shift in renal tissue of mice affected by lupus, we are currently performing longitudinal NAD(P)H fluorescence lifetime imaging of the kidney in collaboration with Prof. Dr. R. Voll (Erlangen, CRC130).

Long-lived plasma cells are central players in forming both physiologic and pathologic immunological memory. Due to a lack of technological tools, these cells could not be investigated over their whole lifetime in their natural environment – the bone marrow of long bones. We therefore developed LIMB: Longitudinal Intravital Microscopy of the femoral Bone marrow. In this way, we were able to monitor plasma cells over 157 days in the deep cavity of murine femurs. This technology has also allowed us  to repeatedly monitor monocytes and macrophages, as well as vasculature remodelling in models of tissue regeneration after bone injury.

Independent of the organ of interest, intravital microscopic investigation faces the challenge of capturing the high complexity of cellular subpopulations and extracellular tissue structures in a dynamic manner. This requires simultaneous multiplexed imaging of the living tissue, as designed and demonstrated by us  imaging germinal center reactions in murine popliteal lymph nodes.

Keywords
Intravital multi-photon microscopy
NAD(P)H metabolism
Functional in vivo imaging

Biophysical Analytics Prof. Dr. rer. nat. Raluca Niesner Phone +49 (0)30 28460-683 niesner@drfz.de more
Continue to Members

Group leader
Prof. Dr. rer. nat. Raluca A. Niesner

Scientists
Dr. Asylkhan Rakhymzhan

PhD students
Ruth Leben
Markus Köhler
Alexander Fiedler
David Reismann

Bachelor/Diploma/Master
Lucie Reuter
Keven Mauersberger

Technicians
Peggy Mex
Robert Günther

Students
Martin Tschaikner
Wjatscheslaw Liublin
Linus Aulich

Continue to Cooperation partners

Volker Andresen, LaVision Biotec – Milteny, Bielefeld/Germany

Dr. Marcus Beutler, APE, Berlin/Germany

Prof. Georg Duda, Julius-Wolf-Institute, Charité /Berlin/Germany

Dr. David Entenberg, Albert Einstein College of Medicine/Intravital Imaging/ New York/US

Prof. Dr. Susanne Hartmann, Freie Universität, Berlin /Veterinary Medicine/Berlin/Germany

Prof. Dr. Anja E. Hauser, DRFZ and Charité/Immune Dynamics/ Berlin/Germany

Prof. Dr. Frank Heppner, Charité /Neuropathology/Berlin/Germany

Dr. Julia Jellusowa, University Freiburg/Faculty for Biology/Freiburg/Germany

Romano Matthys, RISystem, Davos/Switzerland

Prof. Dr. Dirk Mielenz, University Nürenberg-Erlangen / Immunology/ Erlangen/Germany

Prof. Dr. Friedemann Paul, Charité /Neuroimmunology/Berlin/Germany

Dr. Helena Radbruch, Charité /Neuropathology/Berlin/Germany

Prof. Dr. Tim Schulz, DIfE / Potsdam/Germany

Dr. Heinrich Spiecker, LaVision Biotec – Milteny, Bielefeld/Germany

Prof. Dr. Reinhard Voll, University Hospital Freiburg/Rheumatology/Freiburg/Germany

Prof. Dr. Carolina Wählby, Uppsala University/Quantitative Microscopy/Upsalla/Sweden

Prof. Dr. Jürgen Zentek, Freie Universität, Berlin / Veterinary Medicine /Berlin/Germany

Continue to Third party funding projects

DFG: TRR130, C01

DFG: FOR2165-2, TP7

ISAC: ISAC Scholar Collaboration Grant together with Carolina Wählby

Continue to Selected Publications
  1. Synergistic Strategy for Multicolor Two-photon Microscopy: Application to the Analysis of Germinal Center Reactions In Vivo.“ Rakhymzhan A, Leben R, Zimmermann H, Günther R, Mex P, Reismann D, Ulbricht C, Acs A, Brandt AU, Lindquist RL, Winkler TH, Hauser AE, Niesner RA. Sci Rep. 2017 Aug 2;7(1):7101. doi: 10.1038/s41598-017-07165-0.
  2. Analyzing Nicotinamide Adenine Dinucleotide Phosphate Oxidase Activation in Aging and Vascular Amyloid Pathology.“ Radbruch H, Mothes R, Bremer D, Seifert S, Köhler R, Pohlan J, Ostendorf L, Günther R, Leben R, Stenzel W, Niesner RA*, Hauser AE.* Front Immunol. 2017 Jul 31;8:844. doi: 10.3389/fimmu.2017.00844. eCollection 2017.
  3. Longitudinal intravital imaging of the femoral bone marrow reveals plasticity within marrow vasculature.“ Reismann D, Stefanowski J, Günther R, Rakhymzhan A, Matthys R, Nützi R, Zehentmeier S, Schmidt-Bleek K, Petkau G, Chang HD, Naundorf S, Winter Y, Melchers F, Duda G, Hauser AE*, Niesner RA*. Nat Commun. 2017 Dec 18;8(1):2153. doi: 10.1038/s41467-017-01538-9.
  4. Phasor-Based Endogenous NAD(P)H Fluorescence Lifetime Imaging Unravels Specific Enzymatic Activity of Neutrophil Granulocytes Preceding NETosis.“ Leben R, Ostendorf L, van Koppen S, Rakhymzhan A, Hauser AE, Radbruch H, Niesner RA. Int J Mol Sci. 2018 Mar 29;19(4). pii: E1018. doi: 10.3390/ijms19041018.
  5. „Multiplexed fluorescence microscopy reveals heterogeneity among stromal cells in mouse bone marrow sections“ Holzwarth K., Köhler R., Philipsen L., Tokoyoda K., Ladyhina V., Wählby C., Niesner R.A., Hauser A.E. Cytometry Part A, 2018
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