ÿþ<HTML> <HEAD> <TITLE>36th Meeting of the Danish Society for Flow Cytometry - Program and Abstracts</TITLE> </HEAD> <BODY> <H3>36th Meeting of the Danish Society for Flow Cytometry.</H3> <B>Thursday 20 April 2006</B><BR> Auditorium 2, Rigshospitalet,<BR> Blegdamsvej 9, DK-2100 Copenhagen Ø, Denmark.<BR> <OL> <LI><B>Microfluidic Single Cell and Molecule Analysis</B><BR> Scientific Symposium (13:00-16:20).<BR> <P><LI><B>Generalforsamling</B><BR> Annual General Meeting, in Danish (16:30-17:30). </OL> Everyone including members of the Danish Medical Society are welcome.<BR> Registration is not necessary.<BR> <A HREF="http://www.rigshospitalet.dk/rh.nsf/Content/4FB94AAABCEE168EC1256F12004754C5?OpenDocument" TARGET="body">Parkering ved Rigshospitalet</A>. <P>Organizer: <B>Carl-Henrik Brogren</B>, DSFCM Committee (chb@imbg.ku.dk).<BR> <H3>Program and abstracts:</H3> <HR> <B>Microfluidic Single Cell and Molecule Analysis</B> (13:00-16:20). <UL> <LI>13:00-13:10 <B>Carl-Henrik Brogren</B>, DSFCM: Introduction. <P><LI>13:10-13:55 <B>Alexandra Ros</B>, University of Bielefeld, Bielefeld, Germany: <BR>Single Cell Analysis and Particle Sorting in Microfluidic Devices. <P><LI>13:55-14:40 <B>Michael D. Ward et al.</B>, Los Alamos National Laboratory, Los Alamos, New Mexico, USA: <BR>Field-based manipulation of cells and molecules for microfluidic flow cytometry. <P>14:40-15:10 Pause <P><LI>15:10-15:30<B> Anders Wolff</B>, Department of Micro- and Nanotechnology, Technical University of Denmark, Lyngby, Denmark:<BR> Microchip Flow Cytometry with Integrated Polymer Waveguides. <P><LI>15:30-15:50 <B>Ulrik Darling Larsen</B>, Chempaq A/S, Farum, Denmark: <BR>Chempaq XBC blood analyzer. <P><LI>15:50-16:10 <B>Ulrich Krühne</B>, Centre for Microtechnology and Surface Analysis, Danish Technological Institute, Taastrup, Denmark:<BR> The microfluidic activities at the Danish Technological Institute. <P><LI>16:10-16:20 <B>Carl-Henrik Brogren</B>, Department of Medical Biochemistry and Genetics, University of Copenhagen, Copenhagen, Denmark: <BR>Presentation of the new international network group for single cell analysis - go1cell. </UL> <HR> <B>Commercial Exhibitors & Sponsors</B><BR> <UL> <LI><P><A HREF="http://www.bd.com/dk" TARGET="_blank">BD</A>. <LI><A HREF="http://www.corning.com/lifesciences" TARGET="body">Corning Life Sciences</A>. <LI><A HREF="http://www.dms.dk/" TARGET="body">Danish Medical Society</A>. <LI><A HREF="http://www.biotechline.dk/" TARGET="body">Fisher Scientific & Biotech Line</A>. <LI><A HREF="http://www.mengelengineering.dk" TARGET="body">Mengel Engineering</A>. <LI><A HREF="http://www.ramcon.dk/" TARGET="body">Ramcon A/S</A>. <LI><A HREF="http://www.reaction-lab.com" TARGET="body">Reactionlab A/S</A>. </UL> <HR> <B>Generalforsamling</B> (16:30-17:30). <UL> <LI><A HREF="meet36/meet36GFdagsorden.htm" TARGET="body">Dagsorden</A>. </UL> <P><HR> <B>Abstracts</B> <UL> <LI><B>Alexandra Ros</B>, University of Bielefeld, Bielefeld, Germany (Alexandra.Ros@Physik.Uni-Bielefeld.DE): <BR> <B>Single Cell Analysis and Particle Sorting in Microfluidic Devices</B>.<BR> Microfluidic and lab-on-a-chip devices have attracted widespread interest in separation sciences and bioanalysis. Recent designs in microfluidic devices extend common separation concepts by exploiting new phenomena for molecular dynamics at length scales of 10 microns and below, giving rise to novel manipulation tools and non-intuitive phenomena for microseparations. Within this presentation I will focus on two novel concepts for bioseparations based on tailored microfluidic systems. First, a novel single cell microfluidic concept for proteom research and systems nanobiology will be presented comprising single cell navigation and trapping with subsequent on-chip lysis, protein separation and sensitive laser induced fluorescence detection both for labelled and non-labelled proteins. In the second part I will present a paradoxical migration phenomenon based on thermal fluctuations, periodic microstructuring and biased AC electric fields. This phenomenon, termed absolute negative mobility, has recently been demonstrated to occur for colloidal particles in solution. Its size dependent behaviour and thus potential for separation of large biological components, such as organelles and cells will be discussed. <P><LI><B>Michael D. Ward, Gregory Kaduchak, Steve Graves, John C. Martin, Robert Habbersett, James Werner, Peter Goodwin & Anton V. Malko</B>, Los Alamos National Laboratory, Los Alamos, New Mexico, USA (mward@lanl.gov):<BR> <B>Field-based manipulation of cells and molecules for microfluidic flow cytometry</B>.<BR> The flow cell cross-section in a typical conventional flow cytometer is already microfluidic, measuring about 250 by 250 microns. The surrounding systems used for analyzing this cell are anything but micro however, so a great deal of research on  microfluidic cytometry has focused on using integrated chip technology to make the entire package smaller and cheaper. While this is certainly a worthwhile goal, chip technologies also promise to expand the capabilities of conventional flow cytometry by facilitating integration with other powerful lab-on-a-chip technologies. In order to accomplish this integration our labs are developing acoustic, magnetic and electric field based techniques for concentrating, trapping, aligning and sorting cells, particles and molecules. These techniques can be used upstream or downstream of the flow cell. Upstream of the cell, concentration of dilute samples and or enrichment of rare target cells is the primary goal. This sample prep stage expands the range of samples that can be analyzed in microfluidic systems that are typically otherwise confined to analysis of small concentrated volumes. Downstream of the cell, trapping targets of interest that have already been analyzed and or sorted is the primary goal. Trapped targets can be further processed or analyzed on chip, e.g. genetic analysis or immunocytochemistry. If large volumes of sample or sheath are used during flow analysis, this step is particularly important for getting to a small concentrated volume appropriate for whatever process is next. We have specifically demonstrated acoustic and magnetic concentration and trapping of latex particles, human cells, vegetative bacteria and spores. Finally, field based techniques can be used during flow analysis for sorting. We have developed magnetic and electric field devices designed for sorting magnetic particles and single DNA molecules respectively. <P><LI><B>Anders Wolff</B>, Department of Micro- and Nanotechnology, Technical University of Denmark, Lyngby, Denmark (aw@mic.dtu.dk):<BR> <B>Microchip Flow Cytometry with Integrated Polymer Waveguides</B>.<BR> In the Cell Handling Group at MIC  Department for Micro and Nanotechnology (DTU, Lyngby, Denmark) we are working on microfabricated flow cytometers. In this flow cytometers several different optical elements (waveguides, lens and fiber-to-waveguide couplers) are integrated with microfluidic channels to form a complete microchip flow cytometer. All the optical elements, the microfluidic system, and the fiber-to-waveguide couplers were defined in one layer of polymer (SU-8, negative photoresist) by standard photolithography. By using such flowcytometer chips different signals (forward scattering, large angle scattering and fluorescence) were measured simultaneously for bead or cell. As an example rainbow trout red blood cells (RTRBC) and chicken red blood cells (CRBC) with propidium iodide (PI) stained nuclei were measured using the microchip flow cytometer. Because the sizes of both types of cells are close to 3¼m there was little difference in the scattering signals. In contrast, the fluorescence signal was quite different for the two kinds of cells, the fluorescence signals intensities of PI dyed RTRBC are stronger than PI dyed CRBC about 3 fold, which is in agreement with the literature. From 2D plot of the fluorescence and scattered light signal, the two kinds of cells can clearly be distinguished. Such micro flow cytometers can easily be integrated with other microfluidic components and because of the small size several flow cytometer can be integrated on the same chip: Recently we have made a novel microchip with a dielectrophoresis (DEP) selective filter integrated with two micro flow cytometers for real-time monitoring of cell sorting processes. Viable and non-viable yeast cells showed different frequency dependence and were sorted with high efficiency. At 2 MHz, more than 90 % of the viable and less than 10 % of the non-viable cells were captured on the DEP filter. The presented approach provides quantitative real-time data for sorting a large number of cells and will allow optimization of the conditions for, e.g., collecting cancer cells on a DEP filter while normal cells pass through the system. <P><LI><B>Ulrik Darling Larsen</B>, MSc, PhD, Founder, CTO, Chempaq A/S, Hirsemarken 1B, DK-3520 Farum, Denmark (UDL@Chempaq.dk):<BR> <B>Chempaq XBC blood analyzer</B>.<BR> Chempaq has developed a point-of-care haematology analyzer which was launched in Denmark a year ago. The test includes five parameters (Total leukocytes, a three part leukocyte differential, and haemoglobin). The test is performed simply by adding a blood droplet to a cassette called a PAQ and placing it into the apparatus. The apparatus calibrates each cassette and the test is performed in less than 3 minutes. Chempaq s system is the first true point-of-care hematology analyzer with only very little test handling necessary and with no maintenance required. The performance of the system is fully equivalent to other larger and much more expensive analyzers. In December 2005 FDA approved the apparatus as substantially equivalent to current lab standards. Due to the simplicity of the apparatus it is expected that the Chempaq XBC will be granted a CLIA-waiver in 2006 stating that the apparatus is fit for use by nonprofessionals. <P><LI><B>Ulrich Krühne</B>, MSc, PhD, Senior Consultant, Centre for Microtechnology and Surface Analysis, Danish Technological Institute, Gregersensvej 1, Postboks 141, DK 2630 Taastrup, Denmark (Ulrich.Kruhne@teknologisk.dk; www.teknologisk.dk):<BR> <B>The microfluidic activities at the Danish Technological Institute</B>.<BR> The presentation gives a comprehensive overview about the microfluidic activities at the Technological Institute. It illustrates the use of computational fluid dynamic methods for the prediction of microfluidic flow conditions. The examples comprise new laser driven micropumps, micromagnetic separation, tissue engineering examples and some typical examples from the  real world of industry. An open face chip technology is presented with which it is possible to investigate biological matter as e.g. cells under physiological conditions. Experimental results demonstrate the use of the microfluidic platform. The presentation highlights furthermore the special features of laminar flow conditions and introduces to hydrodynamic focussing, the use of thermocouples as mass flow sensors and the experimental results of a simple microfluidic structure for micromagnetic separation of magnetizable beads. <P><LI><B>Carl-Henrik Brogren</B>, University of Copenhagen, Department of Medical Biochemistry and Genetics, Panum Institute, Blegdamsvej 3, DK-2200 Copenhagen, Denmark (chb@imbg.ku.dk):<BR> <B>Presentation of the new international network group for single cell analysis - go1cell</B>.<BR>We are scientists with the research interest focused on the single cell study. Our mission is to perform quantitative structure-to-function molecular analyses on the single cell level by developing novel interdisciplinary research approaches and new micro- and nano-scale technologies. The purpose of the go1cell web site is to facilitate scientific communication between the go1cell networks members related to:<BR> 1. Exchanging research information,<BR> 2. Initiating collaborations with exchange of students and scientists,<BR> 3. Publishing results of our collaborative research,<BR> 4. Raising funds at the international and national level,<BR> 5. Spread of knowledge and technologies in the field of the single cell research by holding meetings and workshops.<BR> Why we are studying the single cell? The scientific problem addressed is that of the nature and extent of the heterogeneity of populations of cells. This heterogeneity is intrinsic to both genetically programmed differentiation and stochastic/epigenetic variation. It is fundamental to cancer, morphogenesis, microbiology and immunology. Due to the lack of appropriate techniques, however, information about it is relatively scarce and the demand for it from academia and industry remains largely unsatisfied. Achieving our objective will constitute a major step forward in satisfying this demand.<BR> What kind of cells we are studying? Bacterial cells, plant cells and animal cells originated from different types of healthy and pathological tissues of variety of species ranging from simple invertebrates to mammals. What are the novel micro- and nan-scale technologies that we are developing? Micro and nano-fluidic devices for separation of single cell molecules. Secondary Ion mass spectrometry SIMS as a tool for quantitative measurements. <BR> What is the impact of our research? Fundamental biology, Medicine, Industry.<BR> Becoming the member. If you or your Laboratory has interests to join go1cell international network group please contact gradimir@gradimir.com or see www.go1cell.net. </UL> <HR> Revised, 6 April 2006 /JKL </BODY> </HTML>