program - spp 1420spp1420.mpikg.mpg.de/data/final program summer school ulm_fine.… ·...

Program:

Summer School 2013

at the University of Ulm

25. - 26. July 2013

Topic: Electron Microscopy

Thursday, 25. July 2013

Welcome address: 09:30-10:00 (lecture room H10)

Introduction of the Central Facility for Electron Microscopy

(Julia Huber)

Morning lectures: 10:00 – 11:45 (lecture room H10)

10:00-10:45

Prof. Dr. Gilles Luquet (UMR BOREA, Biologie des Organismes et Ecosystèmes Aquatiques, Equipe

Evolution des Biominéralisations, Muséum National d'Histoire Naturelle, Paris,

France)

Calcification and calcium storage in Crustaceans: structural and

molecular approaches

10:45-11:00

Coffee break

11:00-11:45

PD Dr. Andreas Ziegler (Central Facility for Electron Microscopy, University of Ulm, Germany)

The effect of physiological saline and organic matrix proteins on shape

and mineral phase of CaCO3 precipitates

11:45-13:00

Lunch break in the mensa

Summer School 2013 – University of Ulm – Electron Microscopy - 1 -


Afternoon lectures: 13:00 – 15:30 (lecture room H10)

13:00-13:45

Dr. Helge-Otto Fabritius (Microstructure Physics and Alloy Design, Max-Planck-Institut für

Eisenforschung, Düsseldorf, Germany)

Structure-property relations in biological materials – Opportunities and

challenges

13:45-14:30

Dr. Erika Griesshaber (Department of Earth- and Environmental Sciences and GeoBioCenter,

LMU Munich, Germany)

Nanoparticle and mesocrystalline domain organization in carbonate

biological hard tissues: Crystal and carbonate phase orientation

patterns obtained from Electron Backscattered Diffraction (EBSD)

14:30-14:45

Coffee break

14:45-15:30

Prof. Dr. sc. nat. (ETH) Paul Walther (Central Facility for Electron Microscopy, University of Ulm, Germany)

Electron microscopy in life science

Summer School 2013 – University of Ulm – Electron Microscopy Summer School 2013 – University of Ulm – Electron Microscopy - 2 -


Visit: 16:00 – 17:45

WITec Wissenschaftliche Instrumente und Technologie GmbH

Lise-Meitner-Str. 6, 89081 Ulm

Scanning Near-field optical Microscopy

Atomic Force Microscopy

Confocal Microscopy

Raman Spectroscopy

Ultrasensitive and fast Raman Imaging

Dinner

18:00-20:00

Dinner in the beer garden of the university

Evening lecture (seminar room of the botanical garden)

20:00-20:45

Prof. Dr. sc. nat. (ETH) Paul Walther

(Central Facility for Electron Microscopy, University of Ulm, Germany)

Electron microscopy in life science: Past, present, future


Friday, 26. July 2013

Lab tour in 4 groups: 08:30-08:45

(Festpunkt M25, room 436, 2nd level [yellow])

methods, sample preparation and demonstrations at the microscopes

FEI 300 kV "Titan" STEM

(STEM tomography)

Hitachi S-5200 FE-SEM

(EDX, etching, polishing, STEM, ultrathin sectioning of non-

demineralized cuticle)

Jeol 1400 TEM

(decalcification, ultrathin sectioning of demineralized cuticle)

Zeiss 962 SEM

(critical point drying, surface structures)

Part I

08:45-09:30

1st round

09:30-10:15

2nd round



10:15-10:30

Coffee break

Morning lecture (lecture room H10)

10:30-11:30

Dr. Martin Friak (Max Planck Institute for Iron Research, Düsseldorf)

Nano-to-macro-scale modeling of hierarchical biocomposites

11:30-12:30

Lunch break in the mensa

Part II (Festpunkt M25, room 436, 2nd level [yellow])

12:30-13:15

3rd round

13:15-14:00

4th round



Visit: 14:30-16:00

Botanical garden of the University of Ulm

(charge 3,00 Euro)

Farewell party: 16:00

(seminar room of the botanical garden)

Drinks and snacks


Abstracts


Calcification and calcium storage in Crustaceans:

Structural and molecular approaches

Prof Dr. Gilles Luquet UMR BOREA, Biologie des Organismes et Ecosystèmes Aquatiques, Equipe

Evolution des Biominéralisations, Muséum National d'Histoire Naturelle, Paris,

France

([email protected])

Crustaceans cyclically replace their cuticle for growing. Most of them harden this

cuticle by sclerotization and calcification. Calcium ions used for this purpose originate

from the water where most of them live and, in a lesser extent, from food.

Nevertheless the terrestrial and some aquatic species use calcium storage strategies.

Crustaceans are then particularly interesting because of their active calcium

metabolism and their ability to form and resorb cyclically (more or less partially) two

kinds of calcified biominerals.

The storage phenomenon is particularly well developed in amphipods, isopods and

decapods. The sites of storage as well as the morphology of the storage deposits are

very diversified. Nevertheless, it seems as a general feature that the precipitate occurs

as calcium carbonate under an amorphous polymorph (ACC).

On our sides, we paid particular attention to an amphipod model, the semi-terrestrial

talitrid Orchestia cavimana, which cyclically stores calcium in two diverticula of the

midgut as calcareous concretions. On the other side, we studied the formation of ACC

deposits by freshwater crayfish in their stomach wall, as so-called gastroliths.

For understanding the formation of these biomineralized structures, we combined

structural and molecular analyses. First, we studied their mineralogical composition by

XRD, FTIR, and Raman spectroscopy and their structure from the macro to the nano-

level by using different chemical treatments and microscopic techniques. Then, the

characterization of molecular components of the organic matrix, focusing on

proteinaceous components and sugars, was performed by biochemistry and molecular

biology.

The sequence analysis of the matrix proteins and the genes encoding these proteins

could lead to the understanding of the strategy used by evolution to built and select

different mineralizing systems.


mailto:[email protected]



The effect of physiological saline and organic matrix

proteins on shape and mineral phase of CaCO3

precipitates

PD Dr. Andreas Ziegler Central Facility for Electron Microscopy, University of Ulm, Germany

([email protected])

Many organisms use calcium carbonate as the inorganic component in skeletal

elements of their body. In Crustacea the mineral phase consists mostly of calcite

and amorphous calcium carbonate (ACC). The amorphous phase is of particular

significance since the crystalline phases are formed from ACC precursors.

Because of its high solubility, amorphous calcium carbonate (ACC) is unstable in

vitro and must be stabilized to prevent spontaneous crystallization. It is known

that in natural systems specific proteins, magnesium and phosphate play a role

in the stabilization, and possibly also in the control of nucleation and short-range

order of biogenic ACC. The relative effects of these components, however, are

poorly understood. Therefore, we conducted experiments that allow to test the

effect of the components of physiological saline and organic matrix proteins on

CaCO3 precipitates. We use the terrestrial isopod Porcellio scaber as a model

which uses ACC for transient storage of cuticular CaCO3 during its moult cycle.

The ACC deposits consist of spherules that are formed in a narrow space, the

ecdysial gap, in a medium whose ionic composition, pH and organic matrix is

under strict control of the hypodermis cells. The presentation will explain the

methods used to 1) measure the cation composition in a narrow space, 2) how

the organic matrix can be extracted, 3) how precipitation experiments are

conducted when only small amounts of organic matrix are available, and 4) how

the mineral phase can be characterised by Raman spectroscopy. Furthermore,

the effect of physiological saline, control protein, and organic matrix proteins of

the deposits on morphology and mineral phase of the precipitates will be

discussed.





Structure-property relations in biological materials –

Opportunities and challenges

Dr. Helge-Otto Fabritius Microstructure Physics and Alloy Design, Max-Planck-Institut für Eisenforschung,

Düsseldorf, Germany

([email protected])

Understanding the structure-property relations and thus the design principles of

biological materials is a valuable source of inspiration for the development and

improvement of synthetic structural materials with tailored properties. Biological

materials are hierarchically structured nano-composites optimized through evolution to

perform vital functions within the specific eco-physiological strains of living organisms.

Most of them consist of a matrix of structural biopolymers like chitin in arthropod

exoskeletons, collagen in vertebrate bones or cellulose in plants and various other

organic and inorganic constituents. Their physical properties are adapted to the

specific functions of the materials and can be very diverse, which is caused by

structural and chemical alterations at different hierarchical levels. Structure,

composition and physical properties of biological materials can be investigated using a

vast variety of constantly improving experimental methods. However, it is still

challenging due to inherent characteristics of the materials like the diversity and small

size of constituents, state of hydration and the many physiological factors influencing

the individual organism. These issues shall be discussed using the crustacean cuticle

as a showcase. The cuticle of crustaceans is a continuous tissue that covers the entire

body and forms the functional exoskeleton of the animals. On the molecular level, the

cuticle consists of chitin and proteins that form fibrils, which are hierarchically

organized over several levels and can be associated with different biominerals, mainly

carbonates and phosphates. On the higher levels of hierarchy, the cuticle forms

skeletal elements with elaborate functions. Investigation of the mechanical properties

of crustacean cuticle showed that the specific design and properties at the nanoscale

contribute significantly to the macroscopic properties. Evidently, the overall properties

depend on the specific microstructure at all levels of hierarchy. However, especially the

properties at small length scales proved to be experimentally hard, if not impossible, to

access due to methodological constraints. Hence, we developed a multiscale model

that can systematically describe and investigate material properties from the atomistic

scale up to the macroscopic level. Through a combination of different modelling

approaches and experimental data, the model can be directly used to translate

biological design principles into material design via virtual prototyping.







Nanoparticle and mesocrystalline domain

organization in carbonate biological hard tissues:

Crystal and carbonate phase orientation patterns

obtained from Electron Backscattered Diffraction

(EBSD)

Dr. Erika Griesshaber Department of Earth- and Environmental Sciences and GeoBioCenter,

LMU Munich, Germany ([email protected])

Mineralized structures generated by biological control are widely recognized as

prototypes for advanced materials. Their exquisite properties are obtained

through the interlinkage of distinct material components and the development of

highly evolved microstructures. Biological hard tissues are hierarchical, where

each hierarchical level contributes to the material property and overall function of

the end-product.

Electron backscatter diffraction (EBSD) is currently one of the best methods

available for the structural characterization of materials (biological and non-

biological), since it provides the ability to study grain morphology,

crystallographic orientation together with the interlinkage of multiple phases.

EBSD is a microdiffraction method and allows the joint analysis of microstructure

evolution and phase distribution. The spatial resolution of EBSD depends on the

system and the sample in question. It is currently in the order of 500 nm for

ceramic materials and 50 nm or better for metals. The accuracy of lattice

parameter measurement, however, is only in the order of 0.1%-1%.

The talk gives first an introduction to the EBSD technique (sample preparation,

data acquisition, data processing and interpretation, spatial and angular

resolution improvements). Subsequently, with case studies on brachiopods,

bivalves and sea urchin teeth the use of EBSD is highlighted for the investigation

of nano-, micro- and macro structures of biological carbonate hard tissues.





Electron microscopy in life science

Prof. Dr. sc. nat. (ETH) Paul Walther Central Facility for Electron Microscopy, University of Ulm, Germany

([email protected])

Electron microscopy allows the investigation of life science samples from

macroscopic to macromolecular resolution. Biological samples usually have a

high water content and are, therefore, not stable in the vacuum of an electron

microscope. They, therefore, need to be dehydrated and cut so that the

information of interest is amenable to the electron beam. For this purpose cryo-

preparation protocols are especially useful, since they allow to prepare the

samples from a defined physiological state. A new emerging field in electron

microscopy is three dimensional imaging. This can be achieved with a number of

different approaches. In this talk three of these methods will be explained in

detail. The traditional method is serial sectioning and imaging the same area in

all sections. Another method is TEM tomography that involves tilting a section

in the electron beam and then reconstruction of the volume by back projection

of the images. When the scanning transmission (STEM) mode is used,

thicker sections (up to 1 µm) can be analyzed. The third approach is focused ion

beam/scanning electron microscopy (FIB/SEM) tomography, in which, a

sample is repeatedly milled with a focused ion beam (FIB) and each newly

produced block face is imaged with the scanning electron microscope (SEM).

This process can be repeated ad libitum in arbitrary small increments allowing

3D analysis of relatively large volumes such as eukaryotic cells. Numerous

examples from cell biology, virology and biological nanostructures will be

presented.





Nano-to-macro-scale modeling of hierarchical

biocomposites

M. Friák1*, M. Petrov1, S. Nikolov2, C. Sachs1, H. Titrian1, U. Aydin1,

A. M. Janus1, H.-O. Fabritius1, L.-F. Zhu1, P. Hemzalová1, D. Ma1,

L. Lymperakis1, D. Raabe1, S. Hild3, A. Ziegler4 and J. Neugebauer1 1Max-Planck-Institut für Eisenforschung GmbH, Düsseldorf, Germany [email protected] 2Institute of Mechanics, Bulgarian Academy of Sciences, Sofia, Bulgaria 3Department of Polymer Science, Johannes Kepler University Linz, Linz, Austria 4Central Facility for Electron Microscopy, University of Ulm, Ulm, Germany

Biological structural materials receive increasing attention by material scientists

because they have been optimized during evolution and they are therefore ideally

suited to study the efficiency of nature's design principles. These materials differ

fundamentally from most man-made structural materials in being structurally

heterogeneous by combining different in/organic constituents into composites with

hierarchical organization. We propose a hierarchical model for the prediction of the

elastic properties of a mineralized arthropod cuticle using ab initio calculations to

find the elastic properties at the nanoscale and employing hierarchical homoge-

nization to find the cuticle properties at all hierarchy levels. Based on our results

we suggest that the mineral-protein matrix possesses a microstructure (so-called

symmetric cell material) which exhibits extremal properties in terms of stiffness. We

also discuss the role of chitin and the multifunctional optimization of the cuticle in

terms of a trade off between stiffness and transport capacity of the pore canal

system (Nikolov et al., Advanced Materials 22 (2010) 519, Nikolov et al., Journal of

the Mechanical Behavior of Biomedical Materials 4 (2011) 129). Recently, we have

further extended our study to analyze the stiffening impact of magnesium additions

on Mg-containing calcite particles (Zhu et al., Journal of the Mechanical Behavior

of Biomedical Materials 20 (2013) 296).



General information

The aim of this summer school…

…is to bring together PhD students, postdocs and senior

scientists to discuss experimental, computational and

theoretical methods applied within the SPP 1420.

Former SPP 1420 members are welcome as well.

Registration…

Please register until the 25. June 2013 at

[email protected]

Travel expenses…

We are dedicated to compensate travel expenses

(accommodation & transportation) for PhD students and

postdocs who belong to the SPP 1420.

Accommodation incl. breakfast should not exceed 72,00

Euro per person per night (only for Ulm).

Senior researchers…

…are very welcome as well but asked to cover travel

expenses via their respective institutions.



General information

Catering...

Lunch: We will have lunch in the mensa of the university. For

SPP1420 members lunch coupons will be provided.

Participants who are not members of the SPP1420 would

have to pay for themselves.

Dinner: On Thursday we will have a barbecue in the beer

garden very close to the botanical garden of the university.

Participants who are not members of the SPP1420 would

have to pay 15 Euro for the barbecue buffet which already

includes 1 drink.


Hotels ibis budget Ulm City (former ETAP HOTEL)

Neutorstrasse 16

89073 ULM

GERMANY

Tel : (+49)73117662720

http://www.etaphotel.com

Hotel Garni Lehrertal

Lehrertalweg 3 (am Eselsberg)

89075 Ulm

Telefon: 0731 954000

Telefax: 0731 95400-50

http://www.lehrertal.de

Hotel Weinstube Zum Bäumle

Kohlgasse 6

89073 Ulm

Telefon: 0731 62287

Telefax: 0731 6022604

http://www.hotel-baeumle.de

Hotel am Rathaus

Kronengasse 8-10

89073 Ulm

Tel: +49 (0)731 - 96849-0

Fax: +49 (0)731 - 96849-49

www.rathausulm.de


http://www.etaphotel.com/

http://www.lehrertal.de/

http://www.hotel-baeumle.de/



http://www.rathausulm.de/

How to find us?

… from ibis budget Ulm City or Hotel Garni Lehrertal (bus stop infront of the

hotels)

Take bus no. 3 (direction „Wissenschaftsstadt“).

Get off at bus stop „Staudingerstraße“, cross the street, cross the parking area,

enter „Eingang Nord“ and enter the first lecture room on the left hand side „H10“.

… from Hotel Weinstube Zum Bäumle

Walk for about 5 min to the bus stop „Theater“. Take bus no. 3 (direction

„Wissenschaftsstadt“).



… from Hotel am Rathaus

Walk for about 2 min to the bus stop „Rathaus“. Take bus no. 6 (direction

„Universität Süd“) until bus stop „Hauptbahnhof“. Take bus no. 3 (direction

„Wissenschaftsstadt“).



The lab tour will take place in the Central Facility for Electron Microscopy

(Universität Ost). It is located in „Festpunkt M25“, room 436 on the second floor

(yellow level).

Via www.ding-ulm.de/‎ you can print out your personal timetable for the bus.


How to find us?


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Coordination and contact for further information

Julia Huber

Zentrale Einrichtung für Elektronenmikroskopie

Universität Ulm

Albert-Einstein-Allee 11 (M25, 436)

Oberer Eselsberg

89069 Ulm

Germany

Tel.: 0731 – 50 – 23441

Email: [email protected] http://www.uniulm.de/einrichtungen/elektronenmikroskopie.html

Updated information about the program is available on the

homepage of the SPP1420: http://spp1420.mpikg.mpg.de/konferenzen/summer-school-2013









http://www.uniulm.de/einrichtungen/elektronenmikroskopie.html

http://spp1420.mpikg.mpg.de/konferenzen/summer-school-2013














Compensation for travel expenses

On the homepage of the SPP1420

(http://spp1420.mpikg.mpg.de/konferenzen/summer-school-

2013/summer-schhol-2013)

you will find a download form for claiming your travel

expenses.

Some important hints:

• Keep all original invoices, train tickets etc.

• The invoice of the hotel should be addressed to

Deutsche Forschungsgemeinschaft

Kennedyallee 40

53175 Bonn

and should also include your name.


http://spp1420.mpikg.mpg.de/konferenzen/summer-school-2013/summer-schhol-2013









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