Medizinische Fakultät Münster


SiCA 2007

4th Münster Conference on Single Cell Analysis 2007
Progress in Research and Technology
Nov. 26-27, 2007

SiCA

Photograph of a macrophage by Lennart Nilsson used by permission.

Scope

The activity of individual cells is the basis of any biological function in living organisms. Advances in analytical technologies allow investigations on the level of single cells and molecules and enhance our understanding of their function and interaction. This conference monitors the current state-of-the-art.


Program
Abstracts

Monday, Nov. 26

Keynote Lecture
13:00 Stephen D. Ginsberg, New York University School of Medicine
Single cell gene expression analysis in human postmortem brain tissues and animal models of neurodegeneration

Session I: Miniaturization & Biosensors
14:00 Jon Cooper, University of Glasgow - cancelled on short notice
Using microfluidics to enable single cell measurements

14:30 Andrea Robitzki, University of Leipzig
Frontiers in nano- and microstructures on microarrays for real time and online biomonitoring of cellular physiology

15:00 Petra Dittrich, ISAS Dortmund
Microfluidic platforms for analysis of living cells and 3D lipid structures

15:30 Coffee Break

Session II: Genomics & Proteomics

16:00 Walter Schubert, University of Magdeburg NBT 2006, 24:1270-1278; Research highlight Nature 2006; Vol 443
The human TOPONOME project: deciphering the protein network code until 2020

16:30 Dietmar Fischer, University of Ulm
New insights into molecular mechanisms underlying axon regeneration through cell sorting

17:00 Markus Veit, Sigma-Aldrich
Single cell whole genome amplification: Unleashing a world within a cell

Tuesday, Nov. 27

Session III: Technologies
08:30 Berenike Maier, University of Münster, research info, more info, SFB629
Regulation and force generation of the DNA import motor of Bacillus subtilis

09:00 Jörg Enderlein, University of Tübingen
Two-focus Fluorescence Correlation Spectroscopy: A versatile for precise measurements of molecular diffusion

09:30 Volker Dötsch, University of Frankfurt
Investigation of conformation and dynamics of proteins inside living cells by in-cell NMR spectroscopy

10:00 Coffee Break

Session IV: Advances in Mass Spectrometry
10:30 Connie Jimenez, University of Amsterdam
From mass spectrometric peptide profiling of single neurons towards imaging mass spectrometry

11:00 Jaume Seuma, University of Sheffield
Fine scale measurement and mapping of cancer biomarkers via laser ablation ICP mass spectrometry

11:30 Keith Compson, Waters Corp.
MALDI Synapt HDMS system: Advances in high-efficiency ion mobility mass spectrometry


Satellite Events

Monday, Nov. 26
10:00 Noushin Delmdahl Sartorius-Stedim Biotech
Being fast and efficient - New technologies for reliable sample preparation.

Sample preparation is always a crucial step when working with biological samples, especially proteins.  It is mandatory to have easy to use and highly reproducible methods in order to achieve scientifically reliable results in down stream applications. In this presentation, we will introduce fast ultrafiltration devices for sample preparation along with passivation methods for increasing recoveries when working with diluted protein samples. For sample fractionation, e.g. prior to 2D-PAGE, an innovative method using ion exchange spin columns based on membrane adsorbers as a matrix will be introduced as a reliable and affordable technology.

11:00 Martin Pieprzyk Fluidigm Europe B.V.
BioMark micro fluidic arrays: A proven method to quantify gene expression within a single cell.

Historically, single-cell gene expression experiments have been difficult and expensive to perform. Fluidigm has introduced a new single-cell gene expression technique that, when used with the BioMark System, produces inexpensive, easily reproducible, gene expression results from single-cell samples. The BioMark system which utilizes micro fluidic technology is ideally suited to single cell work by minimizing reaction size and allowing up to 48 individual reactions to be created from as little as 1ul of sample. The method is ideally suited for high-throughput cell-line studies to determine individual cell behavior in what has been believed to be homozygous populations. The method is also ideally suited to determine single gene cell expression levels in everything from circulating tumor cells (CTCs) to stem cells. The data we will present includes data from single human cells from 8-cell stage embryos which were collected and analyzed for expression of 46 developmental genes.

12:00 Kurt Sieberns Miltenyi Biotec
Gene expression profiling of laser capture microdissected samples using SuperAmp™ amplification method

The analysis of single or a few cells is of increasing importance for the identification of cell-specific marker genes. For example, tumors represent a heterogeneous mixture of neoplastic and non-neoplastic cells. Cells of interest might be represented by a minority of cells and thus, their gene expression profiles might be under the detection limit of conventional methods. The isolation of a certain cell population using laser-assisted microdissection, fluorescent activated cell sorting, or magnetic cell sorting is often a prerequisite to gain insight into cell type-specific gene expression patterns. The SuperAmp™ Service was developed to enable the microarray-based gene expression analysis of low cell numbers, starting with a minimum amount of one cell. The Laser Microdissection and Pressure Catapulting technology (P.A.L.M. Microlaser Technologies) represents a method for e.g. the isolation of single cells from tissue sections. The aim of the present study was to validate the suitability of the SuperAmp™ Technology for laser-capture microdissected cells derived from cryo-fixed and paraffin-embedded melanoma tissue sections. A total of 36 samples were isolated from sections of cryo-fixed or formalin-fixed paraffin-embedded tissue sections. Sample sizes ranged from 1-1000 cells; the sections were excised from the central regions of the melanoma. The gene expression profiling was performed using two-color PIQOR™ Skin Microarrays (Miltenyi Biotec GmbH). The SuperAmp™ Technology offers a robust and reliable method for the analysis of gene expression even of small cell numbers such as cells isolated by laser capture microdissection. The high correlation values support the high reproducibility of the modified global PCR strategy. To our knowledge no comparable method for the microarray analysis of low cell counts is available.

Tuesday, Nov. 27
13:00 Theo Pelzer University of Würzburg
Sex hormones and cardiovascular disease; novel ligands, new hopes and clinical implications

Advances in gender based medicine are inherently linked to our improved understanding of sex hormone effects in the cardiovascular system. The talk will update clinicians and basic scientists in the field of cardiology, gynecology and endocrinology with recent results on the function of newly developed estrogen- and progestins in heart failure, hypertension and cardiac hypertrophy, which have previously been challenged by the negative outcome of clinical trials on the prevention of cardiovascular disease by conventional hormone replacement therapy. The presentation will assist the audience to make their own judgements whether "New ligands" may translate into "New Hopes".

13:45 Monika Stich P.A.L.M Microlaser
Laser microdissection: Building bridges between microscopy and molecular analysis

Laser Microdissection and Pressure Catapulting (LMPC) is a non-contact method to separate cell areas, single cells or subcellular compartments out of a fixed tissue section. In recent investigations this technique was modified and optimized for the work with living cells.
The isolation of individual, e.g. fluorescently marked, cells out of a mixed culture and the subsequent cultivation of these captured cells was established on different cell lines in our lab. After capturing selected cells grow out to new cell colonies, not only preserving their viability and proliferation characteristics, but also keeping their genetic information unaltered. This was proved using LMPC on the carcinoma cell line HCT 116. After re-cultivation the cells were analysed by CGH (comparative genomic hybridization) and M-FISH (multiplex-fluorescence in-situ hybridization).
Further experiments were done with different stem cell lines to present a solution for the challenge of receiving homogenous cell clones. Embryonic stem cells differentiate into various kinds of cells and it is difficult to obtain a specific differentiated cell type from this assay. To overcome this problem, laser-microdissection techniques are the most convenient way to work only with defined stem cells, especially concerning therapy approaches. LMPC offers a method to receive homogenous cell cultures out of one single isolated cell (clonal expansion). In our experiments it was shown that the cells keep their stem cell character, i.e. by detection of different stem cell markers. For example the expression of CD34 and OCT-4 was unchanged after LMPC and re-cultivation. Additionally these results were proved by FACS (Fluorescent Activated Cell Sorting).
The technology of LMPC represents a progress in the contamination-free isolation and analysis of living cells. It opens a wide field of interesting applications in cell and developmental biology and even drug development research, where homogeneous cell clusters or defined clones are needed.


Free for all attendees 

Special issue "Single cell analysis"
Ediitors: Jonathan Sweedler & Edgar Arriaga
Analytical & Bioanalytical Chemistry (2007) 387, 1

Special issue "Proteomics" / "Molecular Imaging"
Editors: Wolf D. Lehmann / Reiner Salzer, ABC (2007) 389, 4

Special issue "Microfluidics in cell analysis"
Editor: Alexandra Ros, ABC (2008) 390

15 % conference discount for Wiley science books!


Registration and Submission

Online registration and abstract submission until Nov. 15, 6 pm.
Later registration is possible on-site at the conference desk.
The deadline for submission of poster presentations is Oct. 15. You will be informed, if your contribution has been accepted.
Thanks to our sponsors registration for external participants is only 45 € (on-site 50 €).
Registration is free for Münster scientists!
We kindly request all participants to register online regardless of whether or not a poster is being presented.
This measure will facilitate organisation of the conference. Conference material will only be available to registered participants.

Sponsors: Please get in touch with Dr. Simone König for requests and registration.


Proceedings

All abstract are published in Biomacromolecular Mass Spectrometry. Furthermore, manuscript submissions on mass spectrometry-related issues are encouraged.

SICA 2004    BMMS 1, #1, 2005, pp 85-97
SICA 2005    BMMS 1, #2, 2007, pp 143-161
SICA 2006    BMMS 1, #3, 2007, pp 193-215
SICA 2007    BMMS 1, #4


Conference Venue

Conference Halls Technologiepark Muenster GmbH, Mendelstr. 11, 48149 Münster, Germany

Muenster, Northrhein-Westphalia, was awarded the LivCom Liveable Community Award in 2004. Plan some time during your stay to see the sights of 1200 years of history!


Organisation

Integrated Functional Genomics
Technology Platform of the Interdisciplinary Center for Clinical Research Münster

  • PD Dr. S. König, IFG
  • Dr. R. Naskar, IZKF

IFG, Röntgenstr. 21, 48149 Münster
IZKF, Albert-Schweitzer-Campus 1, Gebäude D3, 48149 Münster


Gold Sponsors
Waters
SIGMA-ALDRICH
Fluidigm
Sartorius Stedim Biotech

Silver Sponsors
Miltenyi Biotec
Palm
Aura Optik

Bronze Sponsors
Eppendorf
Affymetrix
Keyence
Advalytix
Springer
Agilent Technologies
Bruker Daltronics


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