% IMPORTANT: The following is UTF-8 encoded. This means that in the presence
% of non-ASCII characters, it will not work with BibTeX 0.99 or older.
% Instead, you should use an up-to-date BibTeX implementation like “bibtex8” or
% “biber”.
@PHDTHESIS{Hemonnot:315821,
author = {Hemonnot, Clement},
othercontributors = {Koester, Sarah},
title = {{I}nvestigating {C}ellular {N}anoscale with {X}-{R}ays :
{F}rom {P}roteins to {N}etworks},
school = {University of Göttingen},
type = {Dr.},
reportid = {PUBDB-2016-06090},
pages = {184},
year = {2016},
note = {University of Göttingen, Diss., 2016},
abstract = {The advances and technical improvements of X-ray imaging
techniques, taking advantage of X-ray focussing optics and
high intensity synchrotron sources, nowadays allow for the
use of X-rays to probe the cellular nanoscale. Importantly,
X-rays permit thick samples to be imaged without sectioning
or slicing. In this work, two macromolecules, namely keratin
intermediate filament (IF) proteins and DNA, both essential
components of cells, were studied by X-ray techniques.
Keratin IF proteins make up an integral part of the
cytoskeleton of epithelial cells and form a dense
intracellular network of bundles. This network is built from
monomers in a hierarchical fashion. Thus, the keratin
structure formation spans a large range of length scales
from a few nanometres (monomers) to micrometres (networks).
Here, keratin was studied at three different scales: i)
filaments, ii) bundles and iii) networks. Solution
small-angle X-ray scattering revealed distinct structural
and organisational characteristics of these highly charged
polyelectrolyte filaments, such as increasing radius with
increasing salt concentration and spatial accumulation of
ions depending on the salt concentration. The results are
quantified by employing advanced modelling of keratin IFs by
a core cylinder flanked with Gaussian chains. Scanning
micro-diffraction was used to study keratin at the bundle
scale. Very different morphologies of keratin bundles were
observed at different salt conditions. At the network scale,
new imaging approaches and analyses were applied to the
study of whole cells. Ptychography and scanning X-ray
nano-diffraction imaging were performed on the same cells,
allowing for high resolution in real and reciprocal space,
thereby revealing the internal structure of these networks.
By using a fitting routine based on simulations of IFs
packed on a hexagonal lattice, the radius of each filament
and distance between filaments were retrieved.In mammalian
cells, each nucleus contains 2 nm-thick DNA double helices
with a total length of about 2 m. The DNA strands are packed
in a highly hierarchical manner into individual chromosomes.
DNA was studied in intact cells by visible light microscopy
and scanning X-ray nano-diffraction, unveiling the
compaction und decompaction of DNA during the cell cycle.
Thus, we obtained information on the aggregation state of
the nuclear DNA at a real space resolution on the order of
few hundreds nm. To exploit to the reciprocal space
information, individual diffraction patterns were analysed
according to a generalised Porod’s law at a resolution
down to 10 nm. We were able to distinguish nucleoli,
heterochromatin and euchromatin in the nuclei and follow the
compaction and decompaction during the cell division cycle.},
cin = {DOOR},
cid = {I:(DE-H253)HAS-User-20120731},
pnm = {6G3 - PETRA III (POF3-622) / VH-VI-403 - In-Situ
Nano-Imaging of Biological and Chemical Processes
$(2015_IFV-VH-VI-403)$ / FS-Proposal: I-20130035
(I-20130035) / FS-Proposal: I-20140150 (I-20140150)},
pid = {G:(DE-HGF)POF3-6G3 / $G:(DE-HGF)2015_IFV-VH-VI-403$ /
G:(DE-H253)I-20130035 / G:(DE-H253)I-20140150},
experiment = {EXP:(DE-H253)P-P10-20150101},
typ = {PUB:(DE-HGF)11},
doi = {10.3204/PUBDB-2016-06090},
url = {https://bib-pubdb1.desy.de/record/315821},
}