This document summarizes recent work from the Cremer Lab on superresolution fluorescence microscopy techniques. It discusses: 1) Applications of localization microscopy to image DNA distribution in human cell nuclei at resolutions up to 27,000 DNA sites per square micron and individual human influenza A viruses associated with cell membranes. 2) The 2014 Nobel Prize in Chemistry being awarded to Stefan Hell, a former associate of the Cremer Lab, for his work developing stimulated emission depletion microscopy. 3) Publications from the Cremer Lab in 2014 applying localization microscopy to study chromatin, hepatitis growth factor receptors, and cytomegalovirus proteins.

1. Molecular Constellations 2015
Superresolution Fluorescence Microscopy
C. Cremer
Institute of Molecular Biology (IMB), Mainz/DE
Institute for Pharmacy & Molecular Biotechnology (IPMB)/Kirchhoff-Institute for
Physics (KIP), University Heidelberg
For more information see www.optics.imb-mainz.de
LetiziaMancino
Heidelberg Mainz

2. Superresolution Microscopy* of DNA
Distribution in a human Cell Nucleus.
Intensity Image
after Labelling
with a DNA
specific
Fluorochrome:
~ 5,000 DNA
sites /µm2
Maximum ~ 27,000
DNA sites/µm2
Szczurek et al. (2014) *Localization Microscopy (SPDM)
SPDM 4

3. Superresolution
Microscopy* of
Human
Influenza A
Virus (IAV)
associated with
a human Cell
Membrane
*Localization
Microscopy
(SPDM)
Cremer et al. 2014
Counting of
individual IAV*
along membrane:
N ~ 100
*cluster Ø ~ 120
nm

4. 3 point-like objects in x,y plane with
next neigbour distances 50 nm
a) Labelling with same spectral
signature
b) Labeling with different unique
spectral signatures B,G,R
Computation (scalar Theory):
NA = 1.4, λexc = 488 nm
Optical Isolation by the
“Supernova“ Mode: Induction of
stochastically distributed short
flashes of light emission by
individual moleculesCremer et al. 1996, 1999, 2010, 2011
Principle of Localization Microscopy based Superresolution
(Spectral Precision Distance Microscopy/SPDM)

5. For details see www.optics.imb-mainz.de
Nobel Prize 2014 awarded to former Cremer-
Lab Associate Prof. Stefan Hell
A former associate of the Cremer Lab, Prof. Dr. Stefan
Hell, was awarded the Nobel Prize 2014 in Chemistry
for his work on advanced far field light microscopy.
Three scientists share the award: Two Scientists from
USA, Prof. Eric Betzig and Prof. William Moerner, for
developing single molecule microscopy using
photoactivable proteins, and Prof. Stefan Hell from
Germany, who has been awarded for the development
of Stimulated Emission Depletion Microscopy, a
"focused nanoscopy" method (Scientifc background
given at www.nobelprize.org).

6. Publications Cremer Lab 2014 part I
A.T. Szczurek, K. Prakash, H. K. Lee, D. J. Żurek-Biesiada, G. Best, M.
Hagmann, J. W. Dobrucki, C. Cremer, U. Birk (2014) Single molecule
localization microscopy of the distribution of chromatin using Hoechst and DAPI
fluorescent probes. Nucleus 5 (4): 1-10.
T. Cremer, C. Cremer, P. Lichter (2014) Recollections of a scientific journey
published in Human Genetics: from chromosome territories to interphase
cytogenetics and comparative genome hybridization. Hum Genet (2014)
133:403–416.
Q. Wang, R. Dierkes, R. Kaufmann, C. Cremer (2014) Quantitative analysis of
individual Hepatocyte Growth Factor Receptor Clusters in Influenza A virus
infected human epithelial cells using localization microscopy, Biochimica et
Biophysica Acta 1838: 1191–1198.
C. Cremer, R. Kaufmann, M. Gunkel, F. Polanski, P. Müller, R. Dierkes, S.
Degenhard, C. Wege, M. Hausmann, U. Birk (2014) Application Perspectives of
localization microscopy in Virology. Histochem. Cell. Biol., DOI
10.1007/s00418-014-1203-4.

7. Publications Cremer Lab 2014 part II
P. Müller, N. A. Lemmermann ,• R. Kaufmann , M. Gunkel, D. Paech , G.
Hildenbrand , R. Holtappels, C. Cremer, M. Hausmann (2014) Spatial
distribution and structural arrangement of a murine cytomegalovirus
glycoprotein detected by SPDM localization microscopy. Histochem Cell Biol
DOI 10.1007/s00418-014-1185-2.
C. Cremer (2014) Far‐field Light Microscopy. Encyclopedia of Life Sciences
(eLS), DOI:10.1002/9780470015902.a0005922.pub2.
C. Cremer (2014) Vorstoss in atomare Dimensionen. Laborjournal 7-8/2014: 65
– 67.
A. L. Grab, M. Hagmann, R. Dahint, C. Cremer (2015) Localization microscopy
(SPDM) facilitates high precision control of lithographically produced
nanostructures. Micron 68: 1-7.

8. C. Cremer Lab (2014)
Udo Birk (Deputy GL) Margund Bach
Gerrit Best Zhe Chen
Johann von Hase Florian Schock
Anna Luise Grab Hyun-Keun Lee
Martin Hagmann Ramon Perez
Dongyu Ma Kirti Prakash
Jan Neumann Wladimir Schaufler
Dimka Pieper-Krasteva Mei Yu Chen
Aleksander Szczurek Dominik Waibel
Sabrina Rossberger
Alexander Saroori

9. Collaboration Partners 2014
Heike Allgayer, Univ. Heidelberg/DKFZ
Thomas Cremer, Univ. Muenchen (LMU)
Stefan Dithmar, Univ. Heidelberg/Wiesbaden
Michael Hausmann, Univ. Heidelberg
Reiner Heintzmann, Univ. Jena Peter Huber, DKFZ Heidelberg
Rene Ketting, IMB, Mainz
George Reid, IMB Mainz John Stamatoyannopoulos, Univ. Washington
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