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  • 1.
    Afrasiabi, Roodabeh
    et al.
    KTH, School of Information and Communication Technology (ICT), Materials- and Nano Physics.
    Söderberg, Lovisa M.
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology.
    Joensson, Haakan N.
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology.
    Björk, Per
    Svahn Andersson, Helene
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology.
    Linnros, Jan
    KTH, School of Information and Communication Technology (ICT), Materials- and Nano Physics.
    Integration of a Droplet-Based Microfluidic System and Silicon Nanoribbon FET Sensor2016In: Micromachines, ISSN 2072-666X, E-ISSN 2072-666X, Vol. 7, no 8Article in journal (Refereed)
    Abstract [en]

    We present a novel microfluidic system that integrates droplet microfluidics with a silicon nanoribbon field-effect transistor (SiNR FET), and utilize this integrated system to sense differences in pH. The device allows for selective droplet transfer to a continuous water phase, actuated by dielectrophoresis, and subsequent detection of the pH level in the retrieved droplets by SiNR FETs on an electrical sensor chip. The integrated microfluidic system demonstrates a label-free detection method for droplet microfluidics, presenting an alternative to optical fluorescence detection. In this work, we were able to differentiate between droplet trains of one pH-unit difference. The pH-based detection method in our integrated system has the potential to be utilized in the detection of biochemical reactions that induce a pH-shift in the droplets.

  • 2.
    Ahmadian, Afshin
    et al.
    KTH, School of Biotechnology (BIO), Gene Technology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    AnderssonSvahn, Helene
    KTH, School of Biotechnology (BIO), Nano Biotechnology (closed 20130101). KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Massively parallel sequencing platforms using lab on a chip technologies2011In: Lab on a Chip, ISSN 1473-0197, E-ISSN 1473-0189, Vol. 11, no 16, p. 2653-2655Article in journal (Refereed)
  • 3.
    Ahmadian, Afshin
    et al.
    KTH, Superseded Departments, Biotechnology.
    Russom, Aman
    KTH, Superseded Departments, Biotechnology.
    Andersson, Helene
    KTH, Superseded Departments, Biotechnology.
    Uhlén, Mathias
    KTH, Superseded Departments, Biotechnology.
    Stemme, Göran
    KTH, Superseded Departments, Biotechnology.
    Nilsson, Peter
    KTH, Superseded Departments, Biotechnology.
    SNP analysis by allele-specific extension in a micromachined filter chamber2002In: BioTechniques, ISSN 0736-6205, E-ISSN 1940-9818, Vol. 32, no 4, p. 748-754Article in journal (Refereed)
  • 4.
    Andersson, Helene
    et al.
    KTH, Superseded Departments, Biotechnology.
    Griss, P.
    Stemme, Göran
    KTH, Superseded Departments, Signals, Sensors and Systems.
    Expandable microspheres - surface immobilization techniques2002In: Sensors and actuators. B, Chemical, ISSN 0925-4005, E-ISSN 1873-3077, Vol. 84, no 2-3, p. 290-295Article in journal (Refereed)
    Abstract [en]

    In this study, a novel component for microfluidics is introduced. Expandable microspheres have been studied for their application in microfluidics. Two methods for selective immobilization of expandable microspheres without the use of mechanical barriers on silicon, including patterning by photolithography and self-assembly based on surface chemistry have been shown. After the immobilization step the microspheres were expanded thermally. The expansion is irreversible and the volume of the microspheres increases more than 60 times. Patterns of microspheres with features as small as 15 pm have successfully been generated by photolithography. By using self-assembly the microspheres can conveniently be immobilized in monolayers. Future applications of the expandable microspheres can be as fluidic components, such as one-shot valves or micropumps, positioning other microcomponents or to enlarge the surface area.

  • 5.
    Andersson, Helene
    et al.
    KTH, Superseded Departments, Biotechnology.
    Jonsson, C.
    Moberg, Christina
    KTH, Superseded Departments, Chemistry.
    Stemme, Göran
    KTH, Superseded Departments, Signals, Sensors and Systems.
    Consecutive microcontact printing - ligands for asymmetric catalysis in silicon channels2001In: Sensors and actuators. B, Chemical, ISSN 0925-4005, E-ISSN 1873-3077, Vol. 79, no 1, p. 78-84Article in journal (Refereed)
    Abstract [en]

    Consecutive microcontact printing ( mu CP) has been developed to enable multiple functionalization of silicon surfaces, such as the immobilization of chiral ligands. The technique involves two subsequent printing steps using unstructured poly(methylsiloxane) stamps. The pattern is already defined on the substrate, consisting of etched channels. Hence, no precise alignment is needed between the two printing steps. A carboxylic acid group containing reagent was initially printed onto the silicon oxide surface and transformed to an anhydride. hi the second printing step an ester bond was formed with the hydroxy-functionalized ligand. The formed molecular layers were evaluated by contact angle measurements, scanning electron microscopy (SEM) and electron spectroscopy for chemical analysis (ESCA), indicating that the consecutive mu CP was successful. Initially, printing was performed on planar silicon surfaces but to realize a flow-through microfluidic device for high throughput screening a mu CP technique was developed for etched channels. To verify the technique, hydrophobic valves consisting of octadecyltrichlorosilane were formed using mu CP in deep reactive ion etched channels (50 mum wide and 50 mum deep). The printed hydrophobic patches were visualized by SEM and functioned well. Finally, the consecutive mu CP technique was applied to immobilize the ligand in the channels. The channels were then sealed with a low-temperature bonding technique using an adhesive PDMS film, which does not destroy the printed ligand. In this study mu CP is used in a novel manner. It enables a convenient method for performing complex surface modification of etched structures, which is a frequently appearing problem in biochemical microfluidic systems.

  • 6.
    Andersson, Helene
    et al.
    KTH, Superseded Departments, Biotechnology.
    Jonsson, C.
    Moberg, Christina
    KTH, Superseded Departments, Chemistry.
    Stemme, Göran
    KTH, Superseded Departments, Signals, Sensors and Systems.
    Patterned self-assembled beads in silicon channels2001In: Electrophoresis, ISSN 0173-0835, E-ISSN 1522-2683, Vol. 22, no 18, p. 3876-3882Article in journal (Refereed)
    Abstract [en]

    A novel technique enabling selective bead trapping in microfluidic devices without the use of physical barriers is presented in this paper. It is a fast, convenient and simple method, involving microcontact printing and self-assembly, that can be applied to silicon, quartz or plastic substrates. In the first step, channels are etched in the substrate. The surface chemistry of the internal walls of the channels is then modified by microcontact printing. The chip is submerged in a bead slurry where beads self-assemble based on surface chemistry and immobilize on the internal walls of the channels. Silicon channels (100 mum wide and 50 mum deep) have been covered with monolayers of streptavidin-, amino- and hydroxy-functionalized microspheres and resulted in good surface coverage of beads on the channel walls. A high-resolution pattern of lines of self-assembled streptavidin beads, as narrow as 5 mum, has also been generated on the bottom of a 500 mum wide and 50 mum deep channel. Flow tests were performed in sealed channels with the different immobilized beads to confirm that the immobilized beads could withstand the forces generated by water flowing in the channels. The presented results indicate that single beads can be precisely positioned within microfluidic devices based on self-assembly which is useful as screening and analysis tools within the field of biochemistry and organic chemistry.

  • 7.
    Andersson, Helene
    et al.
    KTH, Superseded Departments, Biotechnology.
    Jonsson, C.
    Moberg, Christina
    KTH, Superseded Departments, Chemistry.
    Stemme, Göran
    KTH, Superseded Departments, Signals, Sensors and Systems.
    Self-assembled and self-sorted array of chemically active beads for analytical and biochemical screening2002In: Talanta: The International Journal of Pure and Applied Analytical Chemistry, ISSN 0039-9140, E-ISSN 1873-3573, Vol. 56, no 2, p. 301-308Article in journal (Refereed)
    Abstract [en]

    A technique for generating a general screening platform consisting of dots of immobilized beads on silicon has been developed via self-sorting and -assembly of different kinds of beads. The dots are defined by a teflon-like film, which due to its hydrophobic characteristics also prevents cross-contamination of liquid from different dots. To enable functionalization of individual dots with different target molecules simultaneously a new way of microcontact printing has been explored where different target solutions are printed in parallel using one stamp. In order to show that this platform can be designed for both biochemical assays and organic chemistry, streptavidin-, amino- and hydroxy-functionalized beads have been self-sorted and -assembled both on separate and common platforms. The self-sorting and -arrangement are based on surface chemistry only, which has not previously been reported. Beads of different sizes and material have successfully been immobilized in line patterns as narrow as 5 mum. Besides silicon, quartz and polyethylene have also been used as substrates.

  • 8.
    Andersson, Helene
    et al.
    KTH. University of Twente, Netherlands.
    Van Berg, A. D.
    From lab-on-a-chip to lab-in-a-cell2005In: Microfluidics, BioMEMS, and Medical Microsystems III, SPIE - International Society for Optical Engineering, 2005, p. 1-12Conference paper (Refereed)
    Abstract [en]

    There are many efforts today trying to mimic the properties of single cells in order to design chips that are as efficient as cells. However, cells are nature's nanotechnology engineering at the scale of atoms and molecules. Therefore, it might be better to vision a microchip that utilizes a single cell as experimentation platform. A novel, so-called Lab-in-a-Cell (LIC) concept is described, where advantage is taken of micro/nanotechnological tools to enable precise control of the biochemical cellular environment and possibility to analyze the composition of single cells. This is followed by a review on the present chip solutions for single cell handling and analysis.

  • 9.
    Andersson, Helene
    et al.
    KTH, School of Electrical Engineering (EES), Signal Processing. University of Twente, Netherlands .
    Van Den Berg, A.
    From LOC to LIC: Using individual cells as experimentation platforms2005In: Nanobiotechnology, ISSN 1551-1286, Vol. 1, no 3, p. 319-321Article in journal (Refereed)
    Abstract [en]

    There are many efforts today trying to mimic the properties of single cells in order to design chips that are as efficient as cells. However, cells are nature's nanotechnology engineering at the scale of atoms and molecules. Therefore, it might be better to vision a microchip that utilizes a single cell as experimentation platform. A novel, so-called Lab-in-a-Cell (LIC) concept is described, where advantage is taken of micro/nanotechnological tools to enable precise control of the biochemical cellular environment and possibility to analyze the composition of single cells.

  • 10.
    Andersson, Helene
    et al.
    KTH, Superseded Departments, Biotechnology.
    van den Berg, A.
    Microfabrication and microfluidics for tissue engineering: state of the art and future opportunities2004In: Lab on a Chip, ISSN 1473-0197, E-ISSN 1473-0189, Vol. 4, no 2, p. 98-103Article, review/survey (Refereed)
    Abstract [en]

    An introductory overview of the use of microfluidic devices for tissue engineering is presented. After a brief description of the background of tissue engineering, different application areas of microfluidic devices are examined. Among these are methods for patterning cells, topographical control over cells and tissues, and bioreactors. Examples where microfluidic devices have been employed are presented such as basal lamina, vascular tissue, liver, bone, cartilage and neurons. It is concluded that until today, microfluidic devices have not been used extensively in tissue engineering. Major contributions are expected in two areas. The first is growth of complex tissue, where microfluidic structures ensure a steady blood supply, thereby circumventing the well-known problem of providing larger tissue structures with a continuous flow of oxygen and nutrition, and withdrawal of waste products. The second, and probably more important function of microfluidics, combined with micro/nanotechnology, lies in the development of in vitro physiological systems for studying fundamental biological phenomena.

  • 11.
    Andersson, Helene
    et al.
    KTH, Superseded Departments, Biotechnology.
    van den Berg, A.
    Microfluidic devices for cellomics: a review2003In: Sensors and actuators. B, Chemical, ISSN 0925-4005, E-ISSN 1873-3077, Vol. 92, no 3, p. 315-325Article, review/survey (Refereed)
    Abstract [en]

    A review of microfluidic devices for cellomics is presented. After a brief description of the historical background of Lab-on-Chip (LOC) devices, different areas are reviewed. Devices for cell sampling are presented, followed by cell trapping and cell sorting devices based upon mechanical and electrical principles. Subsequently, a popular type of cell sorters, flow cytometers, is considered, followed by a chapter describing devices for cell treatment: cell lysis, poration/gene transfection and cell fusion devices. Finally a number of microfluidic devices for cellular studies are reviewed. The large amount of very recent publications treated in this review indicates the rapidly growing interest in this exciting application area of LOC.

  • 12.
    Andersson, Helene
    et al.
    KTH, Superseded Departments, Signals, Sensors and Systems.
    van den Berg, A.
    Microtechnologies and nanotechnologies for single-cell analysis2004In: Current Opinion in Biotechnology, ISSN 0958-1669, E-ISSN 1879-0429, Vol. 15, no 1, p. 44-49Article, review/survey (Refereed)
    Abstract [en]

    Many efforts are currently underway to try and mimic the properties of single cells with the aim of designing chips that are as efficient as cells. However, cells are nature's nanotechnology engineering at the scale of atoms and molecules, and it might be better to envision a microchip that utilizes a single cell as an experimentation platform. A novel, so-called laboratory-in-a-cell concept has been described, where advantage is taken of micro-and nanotechnological tools to enable precise control of the biochemical cellular environment; these tools also offer the possibility to analyse the composition of single cells. Methods for single-cell handling and analysis are being developed and will be required for this concept to progress further.

  • 13.
    Andersson, Helene
    et al.
    KTH, School of Biotechnology (BIO), Nano Biotechnology (closed 20130101).
    van den Berg, Albert
    KTH.
    Where are the biologists?: A series of mini-reviews covering new trends in fundamental and applied research, and potential applications of miniaturised technologies2006In: Lab on a Chip, ISSN 1473-0197, E-ISSN 1473-0189, Vol. 6, no 4, p. 467-470Article in journal (Refereed)
  • 14.
    Andersson, Helene
    et al.
    KTH, Superseded Departments, Biotechnology.
    van der Wijngaart, Wouter
    KTH, Superseded Departments, Signals, Sensors and Systems.
    Enoksson, P.
    Stemme, Göran
    KTH, Superseded Departments, Signals, Sensors and Systems.
    Micromachined flow-through filter-chamber for chemical reactions on beads2000In: Sensors and actuators. B, Chemical, ISSN 0925-4005, E-ISSN 1873-3077, Vol. 67, no 1-2, p. 203-208Article in journal (Refereed)
    Abstract [en]

    A new flow-through micromachined device for chemical reactions on beads has been designed, manufactured, and characterized. The device has an uncomplicated planar design and microfabrication process. Both nonmagnetic and magnetic beads can be collected in the reaction chamber without the use of external magnets. The sample flow-through volume of liquid or gas is adjustable and unlimited. The device is sealed with Pyrex to allow real time optical detection of the chemical reactions. At a constant pressure of 3 kPa at the inlet the flow rate for water is about 3.5 mu l/min without beads in the filter chamber, for all the designs. The smallest reaction chamber has a volume of 0.5 nl and can collect approximately 50 beads with a diameter of 5.50 mu m. At a constant pressure of 3 kPa at the inlet, the flow rate for water is about 2.0 mu l/min when the reaction chamber is completely packed with beads. Hence, the flow rate decreases with about 40% when the reaction chamber is packed with beads. The flow-through microfluidic device is not sensitive to gas bubbles, and clogging of the filter is rare and reversible. The beads are easy to remove from the reaction chamber making the micromachined flow-through device reusable. A new and simple technique for fluid interconnection is developed.

  • 15.
    Andersson, Helene
    et al.
    KTH, Superseded Departments, Biotechnology.
    van der Wijngaart, Wouter
    KTH, Superseded Departments, Signals, Sensors and Systems.
    Griss, P.
    Niklaus, Frank
    KTH, Superseded Departments, Signals, Sensors and Systems.
    Stemme, Göran
    KTH, Superseded Departments, Signals, Sensors and Systems.
    Hydrophobic valves of plasma deposited octafluorocyclobutane in DRIE channels2001In: Sensors and actuators. B, Chemical, ISSN 0925-4005, E-ISSN 1873-3077, Vol. 75, no 1-2, p. 136-141Article in journal (Refereed)
    Abstract [en]

    The suitability of using octafluorocyclobutane (C4F8) patches as hydrophobic valves in microfluidic biochemical applications has been shown. A technique has been developed to generate lithographically defined C4F8 hydrophobic patches in deep reactive ion-etched silicon channels. Some of the advantages of this process are that no specific cleaning of the substrate is required, C4F8 is deposited on the sidewalls and the bottom of the channels, a standard photoresist mask can be used to define the patches, and that it is a fast and convenient dry chemical process performed by a standard inductively coupled plasma etcher using the Bosch process. Different patch lengths (200-1000 mum) of C4F8 were deposited in 50 mum wide channels to evaluate which size is most suitable for microfluidic biochemical applications. The valve function of the hydrophobic patches was tested for the following liquids: DD water, acetone, propanol, bead solution and a mixture used for pyrosequencing of DNA. Patch lengths of 200 mum of C4F8 successfully stopped each solution for at least 20 consecutive times. The C4F8 film resists water for at least 5 h. The hydrophobic valve also resists very high concentrations (25%) of surfactants (Tween 80). C4F8 shows a much higher resistance towards water and surface active solutions than previous hydrophobic patches. However, 50% Tween 80 was not stopped at all by the hydrophobic patch. An applied pressure of 760 Pa at the inlet was needed for water to over-run the hydrophobic patch.

  • 16.
    Andersson, Helene
    et al.
    KTH, Superseded Departments, Biotechnology.
    van der Wijngaart, Wouter
    KTH, Superseded Departments, Signals, Sensors and Systems.
    Nilsson, Peter
    KTH, Superseded Departments, Biotechnology.
    Enoksson, P.
    Stemme, Göran
    KTH, Superseded Departments, Signals, Sensors and Systems.
    A valve-less diffuser micropump for microfluidic analytical systems2001In: Sensors and actuators. B, Chemical, ISSN 0925-4005, E-ISSN 1873-3077, Vol. 72, no 3, p. 259-265Article in journal (Refereed)
    Abstract [en]

    The suitability of valve-less micropumps in biochemistry has been shown. Fluids encountered in various biochemical methods that are problematic for other micropumps have been pumped with good performance. The pump is fabricated as a silicon-glass stack with a new process involving three subsequent deep reactive ion etching steps. Some of the main advantages of the valve-less diffuser pump are the absence of moving parts (excluding the pump diaphragm), the uncomplicated planar design, and high pump performance in terms of pressure head and flow rare. In addition, the micropump is self-priming and insensitive to particles and bubbles present in the pumped media. The results show that the valve-less micropump successfully pumps fluids within the viscosity range of 0.001-0.9 N s/m(2). The micropump is not sensitive to the density, ionic strength, or pH of the pumped media. Effective pumping of solutions containing beads of different sizes was also demonstrated. Living cells were pumped without inducing cell damage and no cell adhesion within the pump chamber was found. No valve-less micropump has previously been reported to pump such a wide variety of fluids.

  • 17.
    Andersson, Helene
    et al.
    KTH, Superseded Departments, Biotechnology.
    van der Wijngaart, Wouter
    KTH, Superseded Departments, Signals, Sensors and Systems.
    Stemme, Göran
    KTH, Superseded Departments, Signals, Sensors and Systems.
    Micromachined filter-chamber array with passive valves for biochemical assays on beads2001In: Electrophoresis, ISSN 0173-0835, E-ISSN 1522-2683, Vol. 22, no 2, p. 249-257Article in journal (Refereed)
    Abstract [en]

    The filter-chamber array presented here enables a real-time parallel analysis of three different samples on beads in a volume of 3 nL, on a 1 cm(2) chip. The filter-chamber array is a system containing three filter-chambers, three passive valves at the inlet channels and a common outlet. The design enables parallel sample handling and time-controlled analysis. The device is microfabricated in silicon and sealed with a Pyrex lid to enable real-time analysis. Single nucleotide polymorphism analysis by using pyrose-quencing has successfully been performed in single filter-chamber devices. The passive valves consist of plasma-deposited octafluorocyclobutane and show a much higher resistance towards water and surface-active solutions than previous hydrophobic patches. The device is not sensitive to gas bubbles, clogging is rare and reversible, and the filter-chamber array is reusable. More complex (bio)chemical reactions on beads can be performed in the devices with passive valves than in the devices without valves.

  • 18.
    Andersson Svahn, Helene
    et al.
    KTH, School of Biotechnology (BIO), Proteomics (closed 20130101). University of Twente, Netherlands .
    Van Den Berg, A.
    Single cells or large populations?2007In: Lab on a Chip, ISSN 1473-0197, E-ISSN 1473-0189, Vol. 7, no 5, p. 544-546Article in journal (Refereed)
  • 19.
    Antypas, H.
    et al.
    Karolinska Inst, Dept Neurosci, Swedish Med Nanosci Ctr, Stockholm, Sweden..
    Veses-Garcia, M.
    Karolinska Inst, Dept Neurosci, Swedish Med Nanosci Ctr, Stockholm, Sweden..
    Weibull, Emelie
    KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science.
    Svahn Andersson, Helene
    KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science, Nano Biotechnology.
    Richter-Dahlfors, A.
    Karolinska Inst, Dept Neurosci, Swedish Med Nanosci Ctr, Stockholm, Sweden..
    A universal platform for selection and high-resolution phenotypic screening of bacterial mutants using the nanowell slide2018In: Lab on a Chip, ISSN 1473-0197, E-ISSN 1473-0189, Vol. 18, no 12, p. 1767-1777Article in journal (Refereed)
    Abstract [en]

    The Petri dish and microtiter plate are the golden standard for selection and screening of bacteria in microbiological research. To improve on the limited resolution and throughput of these methods, we developed a universal, user-friendly platform for selection and high-resolution phenotypic screening based on the nanowell slide. This miniaturized platform has an optimal ratio between throughput and assay complexity, holding 672 nanowells of 500 nl each. As monoclonality is essential in bacterial genetics, we used FACS to inoculate each nanowell with a single bacterium in 15 min. We further extended the protocol to select and sort only bacteria of interest from a mixed culture. We demonstrated this by isolating single transposon mutants generated by a custom-made transposon with dual selection for GFP fluorescence and kanamycin resistance. Optical compatibility of the nanowell slide enabled phenotypic screening of sorted mutants by spectrophotometric recording during incubation. By processing the absorbance data with our custom algorithm, a phenotypic screen for growth-associated mutations was performed. Alternatively, by processing fluorescence data, we detected metabolism-associated mutations, exemplified by a screen for -galactosidase activity. Besides spectrophotometry, optical compatibility enabled us to perform microscopic analysis directly in the nanowells to screen for mutants with altered morphologies. Despite the miniaturized format, easy transition from nano- to macroscale cultures allowed retrieval of bacterial mutants for downstream genetic analysis, demonstrated here by a cloning-free single-primer PCR protocol. Taken together, our FACS-linked nanowell slide replaces manual selection of mutants on agar plates, and enables combined selection and phenotypic screening in a one-step process. The versatility of the nanowell slide, and the modular workflow built on mainstream technologies, makes our universal platform widely applicable in microbiological research.

  • 20.
    Bai, Yunpeng
    et al.
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Weibull, Emilie
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Jönsson, Håkan
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Andersson Svahn, Helene
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Interfacing picoliter droplet microfluidics with addressable microliter compartments using fluorescence activated cell sorting2014In: Sensors and actuators. B, Chemical, ISSN 0925-4005, E-ISSN 1873-3077, Vol. 194, p. 249-254Article in journal (Refereed)
    Abstract [en]

    Droplet microfluidic platforms have, while enabling high-throughput manipulations and the assaying of single cell scale compartments, been lacking interfacing to allow macro scale access to the output from droplet microfluidic operations. Here, we present a simple and high-throughput method for individually directing cell containing droplets to an addressable and macro scale accessible microwell slide for downstream analysis. Picoliter aqueous droplets containing low gelling point agarose and eGFP expressing Escherichia coli (E. coli) are created in a microfluidic device, solidified to agarose beads and transferred into an aqueous buffer. A Fluorescence activated cell sorter (FACS) is used to sort agarose beads containing cells into microwells in which the growth and expansion of cell colonies is monitored. We demonstrate fast sorting and high accuracy positioning of sorted 15 μm gelled droplet agarose beads into microwells (14 × 48) on a 25 mm × 75 mm microscope slide format using a FACS with a 100 μm nozzle and an xy-stage. The interfacing method presented here enables the products of high-throughput or single cell scale droplet microfluidics assays to be output to a wide range of microtiter plate formats familiar to biological researchers lowering the barriers for utilization of these microfluidic platforms.

  • 21.
    Barbe, Laurent
    et al.
    KTH, School of Biotechnology (BIO).
    Lundberg, Emma
    KTH, School of Biotechnology (BIO), Proteomics.
    Oksvold, Per
    KTH, School of Biotechnology (BIO), Proteomics.
    Stenius, Anna
    KTH, School of Biotechnology (BIO).
    Lewin, Erland
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cell Physics.
    Björling, Erik
    KTH, School of Biotechnology (BIO).
    Asplund, Anna
    Department of Genetics and Pathology, Rudbeck Laboratory, Uppsala University.
    Pontén, Fredrik
    Department of Genetics and Pathology, Rudbeck Laboratory, Uppsala University.
    Brismar, Hjalmar
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cell Physics.
    Uhlén, Mathias
    KTH, School of Biotechnology (BIO), Proteomics.
    Andersson-Svahn, Helene
    KTH, School of Biotechnology (BIO), Proteomics.
    Toward a confocal subcellular atlas of the human proteome2008In: Molecular and cellular proteomics, ISSN 1535-9476, Vol. 7, no 3, p. 499-508Article in journal (Refereed)
    Abstract [en]

    Information on protein localization on the subcellular level is important to map and characterize the proteome and to better understand cellular functions of proteins. Here we report on a pilot study of 466 proteins in three human cell lines aimed to allow large scale confocal microscopy analysis using protein-specific antibodies. Approximately 3000 high resolution images were generated, and more than 80% of the analyzed proteins could be classified in one or multiple subcellular compartment(s). The localizations of the proteins showed, in many cases, good agreement with the Gene Ontology localization prediction model. This is the first large scale antibody-based study to localize proteins into subcellular compartments using antibodies and confocal microscopy. The results suggest that this approach might be a valuable tool in conjunction with predictive models for protein localization.

  • 22.
    Björk, S. M.
    et al.
    KTH, Centres, Science for Life Laboratory, SciLifeLab. KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. Novo Nordisk Foundation Center for Biosustainability.
    Sjöström, S. L.
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. Novo Nordisk Foundation Center for Biosustainability.
    Andersson-Svahn, Helene
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. Novo Nordisk Foundation Center for Biosustainability.
    Jönsson, Håkan N.
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. Novo Nordisk Foundation Center for Biosustainability.
    Tuning microfluidic cell culture conditions for droplet based screening by metabolite profiling2015In: MicroTAS 2015 - 19th International Conference on Miniaturized Systems for Chemistry and Life Sciences, Chemical and Biological Microsystems Society , 2015, p. 1377-1379Conference paper (Refereed)
    Abstract [en]

    We investigate the impact of droplet culture conditions on cell metabolic state by determining key metabolite concentrations in S. cerevisiae cultures in different microfluidic droplet culture formats. Control of culture conditions is critical for single cell screening in droplets, as cell metabolic state directly affects production yields in cell factories. Metabolite profiling provides a more nuanced estimate of cell state compared to proliferation studies alone. We show that the choice of droplet incubation format has an impact on cell proliferation and metabolite production. Furthermore, we engineered a new better oxygenated droplet incubation format, with retained droplet stability and size.

  • 23.
    Björk, Sara M.
    et al.
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Sjostrom, Staffan L.
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Andersson-Svahn, Helene
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Jönsson, Håkan N.
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Metabolite profiling of microfluidic cell culture conditions for droplet based screening2015In: Biomicrofluidics, ISSN 1932-1058, E-ISSN 1932-1058, Vol. 9, no 4, article id 044128Article in journal (Refereed)
    Abstract [en]

    We investigate the impact of droplet culture conditions on cell metabolic state by determining key metabolite concentrations in S. cerevisiae cultures in different microfluidic droplet culture formats. Control of culture conditions is critical for single cell/clone screening in droplets, such as directed evolution of yeast, as cell metabolic state directly affects production yields from cell factories. Here, we analyze glucose, pyruvate, ethanol, and glycerol, central metabolites in yeast glucose dissimilation to establish culture formats for screening of respiring as well as fermenting yeast. Metabolite profiling provides a more nuanced estimate of cell state compared to proliferation studies alone. We show that the choice of droplet incubation format impacts cell proliferation and metabolite production. The standard syringe incubation of droplets exhibited metabolite profiles similar to oxygen limited cultures, whereas the metabolite profiles of cells cultured in the alternative wide tube droplet incubation format resemble those from aerobic culture. Furthermore, we demonstrate retained droplet stability and size in the new better oxygenated droplet incubation format.

  • 24.
    Björk, Sara
    et al.
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Sjöström, Staffan L.
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Andersson Svahn, Helene
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Jönsson, Håkan N.
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Controlling cell metabolic state in droplet microfluidicsManuscript (preprint) (Other academic)
  • 25.
    Chinnasamy, Thiruppathiraja
    et al.
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Segerink, Loes I.
    Nystrand, Mats
    Gantelius, Jesper
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Andersson Svahn, Helene
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    A lateral flow paper microarray for rapid allergy point of care diagnostics2014In: The Analyst, ISSN 0003-2654, E-ISSN 1364-5528, Vol. 139, no 10, p. 2348-2354Article in journal (Refereed)
    Abstract [en]

    There is a growing need for multiplexed specific IgE tests that can accurately evaluate patient sensitization profiles. However, currently available commercial tests are either single/low-plexed or require sophisticated instrumentation at considerable cost per assay. Here, we present a novel convenient lateral flow microarray-based device that employs a novel dual labelled gold nanoparticle-strategy for rapid and sensitive detection of a panel of 15 specific IgE responses in 35 clinical serum samples. Each gold nanoparticle was conjugated to an optimized ratio of HRP and anti-IgE, allowing significant enzymatic amplification to improve the sensitivity of the assay as compared to commercially available detection reagents. The mean inter-assay variability of the developed LFM assay was 12% CV, and analysis of a cohort of clinical samples (n = 35) revealed good general agreement with ImmunoCAP, yet with a varying performance among allergens (AUC = [0.54-0.88], threshold 1 kU). Due to the rapid and simple procedure, inexpensive materials and read-out by means of a consumer flatbed scanner, the presented assay may provide an interesting low-cost alternative to existing multiplexed methods when thresholds > 1 kU are acceptable.

  • 26.
    Chinnasamy, Thiruppathiraja
    et al.
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Segerink, Loes I.
    Nystrand, Mats
    Gantelius, Jesper
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Andersson Svahn, Helene
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Point-of-Care Vertical Flow Allergen Microarray Assay: Proof of Concept2014In: Clinical Chemistry, ISSN 0009-9147, E-ISSN 1530-8561, Vol. 60, no 9, p. 1209-1216Article in journal (Refereed)
    Abstract [en]

    BACKGROUND: Sophisticated equipment, lengthy protocols, and skilled operators are required to perform protein microarray-based affinity assays. Consequently, novel tools are needed to bring biomarkers and biomarker panels into clinical use in different settings. Here, we describe a novel paper-based vertical flow microarray (VFM) system with a multiplexing capacity of at least 1480 microspot binding sites, colorimetric readout, high sensitivity, and assay time of < 10 min before imaging and data analysis. METHOD: Affinity binders were deposited on nitrocellulose membranes by conventional microarray printing. Buffers and reagents were applied vertically by use of a flow controlled syringe pump. As a clinical model system, we analyzed 31 precharacterized human serum samples using the array system with 10 allergen components to detect specific IgE reactivities. We detected bound analytes using gold nanoparticle conjugates with assay time of <= 10 min. Microarray images were captured by a consumer-grade flatbed scanner. RESULTS: A sensitivity of 1 ng/mL was demonstrated with the VFM assay with colorimetric readout. The reproducibility (CV) of the system was < 14%. The observed concordance with a clinical assay, Immuno-CAP, was R-2 = 0.89 (n = 31). CONCLUSIONS: In this proof-of-concept study, we demonstrated that the VFM assay, which combines features from protein microarrays and paper-based colorimetric systems, could offer an interesting alternative for future highly multiplexed affinity point-of-care testing.

  • 27. DaCosta, R. S.
    et al.
    Andersson, Helene
    KTH, School of Biotechnology (BIO), Nano Biotechnology.
    Cirocco, M.
    Marcon, N. E.
    Wilson, B. C.
    Autofluorescence characterisation of isolated whole crypts and primary cultured human epithelial cells from normal, hyperplastic, and adenomatous colonic mucosa2005In: Journal of Clinical Pathology, ISSN 0021-9746, E-ISSN 1472-4146, Vol. 58, no 7, p. 766-774Article in journal (Refereed)
    Abstract [en]

    Background/Aims: In vivo autofluorescence endoscopic imaging and spectroscopy have been used to detect and differentiate benign ( hyperplastic) and preneoplastic ( adenomatous) colonic lesions. This fluorescence is composed of contributions from the epithelium, lamina propria, and submucosa. Because epithelial autofluorescence in normal and diseased tissues is poorly understood, this was the focus of the present study. Methods: Whole colonic crypts were isolated, and short term primary cultures of epithelial cells were established from biopsies of normal, hyperplastic, and adenomatous colon. Autofluorescence ( 488 nm excitation) was examined by confocal fluorescence microscopy. Fluorescently labelled organelle probes and transmission electron microscopy were used to identify subcellular sources of fluorescence. Results: Mitochondria and lysosomes were identified as the main intracellular fluorescent components in all cell types. Normal and hyperplastic epithelial cells were weakly autofluorescent and had similar numbers of mitochondria and lysosomes, whereas adenomatous ( dysplastic) epithelial cells showed much higher autofluorescence, and numerous highly autofluorescent lysosomal ( lipofuscin) granules. Conclusions: Short term primary cell cultures from endoscopic biopsies provide a novel model to understand differences in colonic tissue autofluorescence at the glandular ( crypt) and cellular levels. The differences between normal, hyperplastic, and adenomatous epithelial cells are attributed in part to differences in the intrinsic numbers of mitochondria and lysosomes. This suggests that the detection of colonic epithelial fluorescence alone, if possible, may be sufficient to differentiate benign ( hyperplastic) from preneoplastic and neoplastic ( adenomatous) colonic intramucosal lesions during in vivo fluorescence endoscopy. Furthermore, highly orange/red autofluorescent intracellular granules found only in dysplastic epithelial cells may serve as a potential biomarker.

  • 28. DaCosta, R. S.
    et al.
    Andersson, Helene
    KTH, Superseded Departments, Biotechnology.
    Wilson, B. C.
    Molecular fluorescence excitation-emission matrices relevant to tissue spectroscopy2003In: Photochemistry and Photobiology, ISSN 0031-8655, E-ISSN 1751-1097, Vol. 78, no 4, p. 384-392Article in journal (Refereed)
    Abstract [en]

    In vivo and ex vivo studies of fluorescence from endogenous and exogenous molecules in tissues and cells are common for applications such as detection or characterization of early disease. A systematic determination of the excitation-emission matrices (EEM) of known and putative endogenous fluorophores and a number of exogenous fluorescent photodynamic therapy drugs has been performed in solution. The excitation wavelength range was 250-520 nm, with fluorescence emission spectra collected in the range 260-750 nm. In addition, EEM of intact normal and adenomatous human colon tissues are presented as an example of the relationship to the EEM of constituent fluorophores and illustrating the effects of tissue chromophore absorption. As a means to make this large quantity of spectral data generally available, an interactive database has been developed. This currently includes EEM and also absorption spectra of 35 different endogenous and exogenous fluorophores and chromophores and six photosensitizing agents. It is intended to maintain and extend this database in the public domain, accessible through the Photochemistry and Photobiology website (http://www.aspjournal.com/).

  • 29. Di Carlo, Dino
    et al.
    Huang, Yanyi
    Andersson-Svahn, Helene
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Emerging investigators: new challenges spawn new innovations2014In: Lab on a Chip, ISSN 1473-0197, E-ISSN 1473-0189, Vol. 14, no 15, p. 2599-2600Article in journal (Refereed)
  • 30.
    Dias, Jorge T.
    et al.
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Lama, Lara
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Gantelius, Jesper
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Andersson-Svahn, Helene
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Minimizing antibody cross-reactivity in multiplex detection of biomarkers in paper-based point-of-care assays2016In: Nanoscale, ISSN 2040-3364, E-ISSN 2040-3372, Vol. 8, no 15, p. 8195-8201Article in journal (Refereed)
    Abstract [en]

    Highly multiplexed immunoassays could allow convenient screening of hundreds or thousands of protein biomarkers simultaneously in a clinical sample such as serum or plasma, potentially allowing improved diagnostic accuracy and clinical management of many conditions such as autoimmune disorders, infections, and several cancers. Currently, antibody microarray-based tests are limited in part due to cross reactivity from detection antibody reagents. Here we present a strategy that reduces the cross-reactivity between nanoparticle-bound reporter antibodies through the application of ultrasound energy. By this concept, it was possible to achieve a sensitivity 10(3)-fold (5 pg mL(-1)) lower than when no ultrasound was applied (50 ng mL(-1)) for the simultaneous detection of three different antigens. The detection limits and variability achieved with this technique rival those obtained with other types of multiplex sandwich assays.

  • 31.
    Dias, Jorge. T.
    et al.
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Svedberg, Gustav
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Nystrand, M.
    Svahn Andersson, Helene
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Gantelius, Jesper
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Rapid nanoprobe signal enhancement by in situ gold nanoparticle synthesis2018In: Journal of Visualized Experiments, ISSN 1940-087X, E-ISSN 1940-087X, Vol. 2018, no 133, article id e57297Article in journal (Refereed)
    Abstract [en]

    The use of nanoprobes such as gold, silver, silica or iron-oxide nanoparticles as detection reagents in bioanalytical assays can enable high sensitivity and convenient colorimetric readout. However, high densities of nanoparticles are typically needed for detection. The available synthesis-based enhancement protocols are either limited to gold and silver nanoparticles or rely on precise enzymatic control and optimization. Here, we present a protocol to enhance the colorimetric readout of gold, silver, silica, and iron oxide nanoprobes. It was observed that the colorimetric signal can be improved by up to a 10000-fold factor. The basis for such signal enhancement strategies is the chemical reduction of Au3+ to Au0. There are several chemical reactions that enable the reduction of Au3+ to Au0. In the protocol, Good's buffers and H2O2 are used and it is possible to favor the deposition of Au0 onto the surface of existing nanoprobes, in detriment of the formation of new gold nanoparticles. The protocol consists of the incubation of the microarray with a solution consisting of chloroauric acid and H2O2 in 2-(N-morpholino)ethanesulfonic acid pH 6 buffer following the nanoprobe-based detection assay. The enhancement solution can be applied to paper and glass-based sensors. Moreover, it can be used in commercially available immunoassays as demonstrated by the application of the method to a commercial allergen microarray. The signal development requires less than 5 min of incubation with the enhancement solution and the readout can be assessed by naked eye or low-end image acquisition devices such as a table-top scanner or a digital camera. 

  • 32.
    Dias, Jorge T.
    et al.
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Svedberg, Gustav
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Nystrand, Mats
    Thermo Fisher Sci IDD, Global Res & Dev, Uppsala, Sweden..
    Svahn Andersson, Helene
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Gantelius, Jesper
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Rapid signal enhancement method for nanoprobe-based biosensing (vol 7, 2017)2018In: Scientific Reports, ISSN 2045-2322, E-ISSN 2045-2322, Vol. 8, no 1, article id 8184Article in journal (Refereed)
    Abstract [en]

    In the Methods section of this Article references 18 to 22 are incorrectly cited. The correct references were omitted from the reference list and appear below as references 1-5. References 18 to 22 are correctly cited in Introduction and Results and Discussion sections. "AuNPs of 10 nm in diameter were prepared following the protocol described by Bastus et al.18." should read: "AuNPs of 10 nm in diameter were prepared following the protocol described by Bastus et al.1." "AgNPs of 90 nm in diameter were prepared following the protocol described by Rivero et al.19." should read: "AgNPs of 90 nm in diameter were prepared following the protocol described by Rivero et al.2" "The size was determined by UV-Vis spectroscopy according to the AgNPs size theory demonstrated by Malynych20." should read: "The size was determined by UV-Vis spectroscopy according to the AgNPs size theory demonstrated by Malynych3." "The coupling of antibody to the NPs was prepared following a modified version of a protocol previously reported by Puertas et al.21." should read: "The coupling of antibody to the NPs was prepared following a modified version of a protocol previously reported by Puertas et al.4." "Microarrays were prepared as previously reported by our group22." should read: "Microarrays were prepared as previously reported by our group5.

  • 33. Douagi, Anna Sjöström
    et al.
    Svahn, Helene Andersson
    KTH, School of Biotechnology (BIO), Proteomics.
    Young researchers to tackle future Grand Challenges2012In: Lab on a Chip, ISSN 1473-0197, E-ISSN 1473-0189, Vol. 12, no 4, p. 680-683Article in journal (Other academic)
  • 34. Eissler, N.
    et al.
    Mao, Y.
    Brodin, D.
    Reuterswärd, Philippa
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology.
    Svahn Andersson, Helene
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology.
    Johnsen, J. I.
    Kiessling, R.
    Kogner, P.
    Combination Therapy of Anti-PD-1 Antibody and CSF-1R Inhibitor Reverses Induction of Suppressive Myeloid Cells and Controls Spontaneous Neuroblastoma Progression2016In: Pediatric Blood & Cancer, ISSN 1545-5009, E-ISSN 1545-5017, Vol. 63, p. S28-S28Article in journal (Refereed)
  • 35. Eissler, Nina
    et al.
    Mao, Yumeng
    Brodin, David
    Reuterswärd, Philippa
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology.
    Svahn Andersson, Helene
    KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology.
    Johnsen, John Inge
    Kiessling, Rolf
    Kogner, Per
    Regulation of myeloid cells by activated T cells determines the efficacy of PD-1 blockade2016In: Oncoimmunology, ISSN 2162-4011, E-ISSN 2162-402X, Vol. 5, no 12, article id e1232222Article in journal (Refereed)
    Abstract [en]

    Removal of immuno-suppression has been reported to enhance antitumor immunity primed by checkpoint inhibitors. Although PD-1 blockade failed to control tumor growth in a transgenic murine neuroblastoma model, concurrent inhibition of colony stimulating factor 1 receptor (CSF-1R) by BLZ945 reprogrammed suppressive myeloid cells and significantly enhanced therapeutic effects. Microarray analysis of tumor tissues identified a significant increase of T-cell infiltration guided by myeloid cell-derived chemokines CXCL9, 10, and 11. Blocking the responsible chemokine receptor CXCR3 hampered T-cell infiltration and reduced antitumor efficacy of the combination therapy. Multivariate analysis of 59 immune-cell parameters in tumors and spleens detected the correlation between PD-L1-expressing myeloid cells and tumor burden. In vitro, anti-PD-1 antibody Nivolumab in combination with BLZ945 increased the activation of primary human T and NK cells. Importantly, we revealed a previously uncharacterized pathway, in which T cells secreted M-CSF upon PD-1 blockade, leading to enhanced suppressive capacity of monocytes by upregulation of PD-L1 and purinergic enzymes. In multiple datasets of neuroblastoma patients, gene expression of CD73 correlated strongly with myeloid cell markers CD163 and CSF-1R in neuroblastoma tumors, and associated with worse survival in high-risk patients. Altogether, our data reveal the dual role of activated T cells on myeloid cell functions and provide a rationale for the combination therapy of anti-PD-1 antibody with CSF-1R inhibitor.

  • 36. Emmelkamp, J.
    et al.
    Gardeniers, J. G. E.
    Andersson, Helene
    KTH, School of Electrical Engineering (EES), Signal Processing. University of Twente, Netherlands .
    Van Den Berg, A.
    Planar nanoneedles on-chip for intracellular measurements2005In: Micro Total Analysis Systems - Proceedings of MicroTAS 2005 Conference: 9th International Conference on Miniaturized Systems for Chemistry and Life Sciences, Transducers Research Foundations, 2005, Vol. 1, p. 400-402Conference paper (Refereed)
    Abstract [en]

    We present for the first time a functional planar SiN-nanoneedle system for intracellular mass transport and in vivo electrophysiological measurements on-chip. Though several micro- or nanoneedles for cell research have been described in literature, no needles of this small size equipped with nanosized inner channels or electrodes have been reported. A propidium iodide assay verifies the excellent penetration performance of the nanoneedles with diminished leakage from the cell after insertion and release of the needle from HL60-cells. Hollow needles connected to on-chip sub-picoliter electrochemical dosing systems are in development.

  • 37. Emmelkamp, J.
    et al.
    Wolbers, F.
    Andersson, Helene
    KTH, Superseded Departments, Biotechnology.
    DaCosta, R. S.
    Wilson, B. C.
    Vermes, I.
    van den Berg, A.
    The potential of autofluorescence for the detection of single living cells for label-free cell sorting in microfluidic systems2004In: Electrophoresis, ISSN 0173-0835, E-ISSN 1522-2683, Vol. 25, no 21-22, p. 3740-3745Article in journal (Refereed)
    Abstract [en]

    A novel method for studying unlabeled living mammalian cells based on their autofluorescence (AF) signal in a prototype microfluidic device is presented. When combined, cellular AF detection and microfluidic devices have the potential to facilitate high-throughput analysis of different cell populations. To demonstrate this, unlabeled cultured cells in microfluidic devices were excited with a 488 nm excitation light and the AF emission (> 505 nm) was detected using a confocal fluorescence microscope (CFM). For example, a simple microfluidic three-port glass microstructure was used together with conventional electroosmotic flow (EOF) to switch the direction of the fluid flow. As a means to test the potential of AF-based cell sorting in this microfluidic device, granulocytes were successfully differentiated from human red blood cells (RBCs) based on differences in AF This study demonstrated the use of a simple microfabricated device to perform high-throughput live cell detection and differentiation without the need for cell-specific fluorescent labeling dyes and thereby reducing the sample preparation time. Hence, the combined use of microfluidic devices and cell AF may have many applications in single-cell analysis.

  • 38.
    Friedman, Mikaela
    et al.
    KTH, School of Biotechnology (BIO), Molecular Biotechnology.
    Lindström, Sara
    KTH, School of Biotechnology (BIO), Nano Biotechnology.
    Andersson-Svahn, Helene
    KTH, School of Biotechnology (BIO), Nano Biotechnology.
    Brismar, Hjalmar
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cell Physics.
    Ståhl, Stefan
    KTH, School of Biotechnology (BIO), Molecular Biotechnology.
    Engineering and characterization of a bispecific HER2 × EGFR-binding affibody molecule2009In: Biotechnology and applied biochemistry, ISSN 0885-4513, E-ISSN 1470-8744, Vol. 54, p. 121-131Article in journal (Refereed)
    Abstract [en]

    HER2 (human epidermal-growth-factor receptor-2; ErbB2) and EGFR (epidermal-growth-factor receptor) are overexpressed in various forms of cancer, and the co-expression of both HER2 and EGFR has been reported in a number of studies. The simultaneous targeting of HER2 and EGFR has been discussed as a strategy with which to potentially increase efficiency and selectivity in molecular imaging and therapy of certain cancers. In an effort to generate a molecule capable of bispecifically targeting HER2 and EGFR, a gene fragment encoding a bivalent HER2-binding affibody molecule was genetically fused in-frame with a bivalent EGFR-binding affibody molecule via a (G(4)S)(3) [(Gly(4)-Ser)(3)]-encoding gene fragment. The encoded 30 kDa affibody construct (Z(HER2))(2)-(G(4)S)(3)-(Z(EGFR))(2), with potential for bs (bispecific) binding to HER2 and EGFR, was expressed in Escherichia coli and characterized in terms of its binding capabilities. The retained ability to bind HER2 and EGFR separately was demonstrated using both biosensor technology and flow-cytometric analysis, the latter using HER2- and EGFR-overexpressing cells. Furthermore, simultaneous binding to HER2 and EGFR was demonstrated in: (i) a sandwich format employing real-time biospecific interaction analysis where the bs affibody molecule bound immobilized EGFR and soluble HER2; (ii) immunofluorescence microscopy, where the bs affibody molecule bound EGFR-overexpressing cells and soluble HER2; and (iii) a cell-cell interaction analysis where the bs affibody molecule bound HER2-overexpressing SKBR-3 cells and EGFR-overexpressing A-431 cells. This is, to our knowledge, the first reported bs affinity protein with potential ability for the simultaneous targeting of HER2 and EGFR. The potential future use of this and similar constructs, capable of bs targeting of receptors to increase the efficacy and selectivity in imaging and therapy, is discussed.

  • 39.
    Frisk, Thomas
    et al.
    KTH, School of Electrical Engineering (EES), Microsystem Technology.
    Rydholm, Susanna
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cell Physics.
    Andersson, Helene
    KTH, School of Electrical Engineering (EES), Microsystem Technology.
    Brismar, Hjalmar
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cell Physics.
    Stemme, Göran
    KTH, School of Electrical Engineering (EES), Microsystem Technology.
    Cultivation of COS7-cells using extracellular matrix in 3D microfluidic surface enlarged structure2005In: Micro Total Analysis Systems 2004, 2005, no 297, p. 261-263Conference paper (Refereed)
    Abstract [en]

    This paper presents a novel method to cultivate cells in a controlled 3D surface enlarged micro-environment. The 3D environment is achieved by insertion of a gel-like extracellular matrix (ECM) mixed with cells into a micromachined silicon fluid structure. Shrinking of the gel enables further flow through the channel. Due to the structure design the shrinking is non-uniform, which results in an increased surface area. With the proposed design cells are alive and viable after 72 h of incubation within the chip.

  • 40.
    Frisk, Thomas
    et al.
    KTH, School of Electrical Engineering (EES), Microsystem Technology.
    Rydholm, Susanna
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cell Physics.
    Andersson, Helene
    KTH, School of Electrical Engineering (EES), Microsystem Technology.
    Brismar, Hjalmar
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cell Physics.
    Stemme, Göran
    KTH, School of Electrical Engineering (EES), Microsystem Technology.
    Three dimensional asymmetric microenvironment for cell biological studies2005In: Proceedings of µTAS 2005 Conference, 2005, p. 915-917Conference paper (Refereed)
    Abstract [en]

    We report on a method for three-dimensional cultivation of cells in a microstructured asymmetric environment. In the system an asymmetric environment is created by using diffusion through a gel of extra cellular matrix proteins surrounded by microfluidic flow channels. Individual cells embedded in the gel react on the concentration gradient. The system has been evaluated both for diffusion properties and based on the cellular response.

  • 41.
    Frisk, Thomas
    et al.
    KTH, School of Electrical Engineering (EES), Microsystem Technology.
    Rydholm, Susanna
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cell Physics.
    Andersson, Helene
    KTH, School of Electrical Engineering (EES), Microsystem Technology.
    Stemme, Göran
    KTH, School of Electrical Engineering (EES), Microsystem Technology.
    Brismar, Hjalmar
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cell Physics.
    A concept for miniaturized 3-D cell culture using an extracellular matrix gel2005In: Electrophoresis, ISSN 0173-0835, E-ISSN 1522-2683, Vol. 26, no 24, p. 4751-4758Article in journal (Refereed)
    Abstract [en]

    This paper presents a novel method to embed, anchor, and cultivate cells in a controlled 3-D flow-through microenvironment. This is realized using an etched silicon pillar flow chamber filled with extracellular matrix (ECM) gel mixed with cells. At 4 degrees C, while in liquid form, ECM gel is mixed with cells and injected into the chamber. Raising the temperature to 37 degrees C results in a gel, with cells embedded. The silicon pillars both stabilize and increase the surface to volume ratio of the gel. During polymerization the gel shrinks, thus creating channels, which enables perfusion through the chip. The pillars increase the mechanical stability of the gel permitting high surface flow rates without surface modifications. Within the structure cells were still viable and proliferating after 6 days of cultivation. Our method thus makes it possible to perform medium- to long-term cultivation of cells in a controlled 3-D environment. This concept opens possibilities to perform studies of cells in a more physiological environment compared to traditional 2-D cultures on flat substrates.

  • 42.
    Frisk, Thomas
    et al.
    KTH, School of Electrical Engineering (EES), Microsystem Technology.
    Rydholm, Susanna
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cell Physics.
    Andersson Svahn, Helene
    KTH, School of Biotechnology (BIO), Nano Biotechnology.
    Brismar, Hjalmar
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cell Physics.
    Stemme, Göran
    KTH, School of Electrical Engineering (EES), Microsystem Technology.
    Three dimensional asymmetric microenvironment for cell biologigal studies2005In: Proceedings Micro Total Analysis Systems (muTAS) 2005, 2005Conference paper (Refereed)
  • 43.
    Frisk, Thomas
    et al.
    KTH, School of Electrical Engineering (EES), Microsystem Technology.
    Rydholm, Susanna
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cell Physics.
    Liebmann, Thomas
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cell Physics.
    Andersson-Svahn, Helene
    KTH, School of Biotechnology (BIO), Nano Biotechnology.
    Stemme, Göran
    KTH, School of Electrical Engineering (EES), Microsystem Technology.
    Brismar, Hjalmar
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cell Physics.
    A microfluidic device for parallel 3-D cell cultures in asymmetric environments2007In: Electrophoresis, ISSN 0173-0835, E-ISSN 1522-2683, Vol. 28, no 24, p. 4705-4712Article in journal (Refereed)
    Abstract [en]

    We demonstrate a concept for how a miniaturized 3-D cell culture in biological extracellular matrix (ECM) or synthetic gels bridges the gap between organ-tissue culture and traditional 2-D cultures. A microfluidic device for 3-D cell culture including microgradient environments has been designed, fabricated, and successfully evaluated. In the presented system stable diffusion gradients can be generated by application of two parallel fluid flows with different composition against opposite sides of a gel plug with embedded cello. Culture for up to two weeks was performed showing cells still viable and proliferating. The cell tracer dye calcein was used to verify gradient formation as the fluorescence intensity in exposed cells was proportional to the position in the chamber. Cellular response to an applied stimulus was demonstrated by use of an adenosine triphosphate gradient where the onset of a stimulated intracellular calcium release also depended on cell position.

  • 44.
    Gantelius, Jesper
    et al.
    KTH, School of Biotechnology (BIO), Nano Biotechnology (closed 20130101).
    Bass, Tarek
    KTH, School of Biotechnology (BIO), Molecular Biotechnology (closed 20130101).
    Gundberg, Anna
    KTH, School of Biotechnology (BIO), Proteomics (closed 20130101).
    Sundberg, Mårten
    KTH, School of Biotechnology (BIO), Proteomics (closed 20130101).
    Sjöberg, Ronald
    KTH, School of Biotechnology (BIO), Proteomics (closed 20130101).
    Nilsson, Peter
    KTH, School of Biotechnology (BIO), Proteomics (closed 20130101).
    Andersson-Svahn, Helene
    KTH, School of Biotechnology (BIO), Nano Biotechnology (closed 20130101).
    A ten-minute high density lateral flow protein microarray assay2011In: 15th International Conference on Miniaturized Systems for Chemistry and Life Sciences 2011: (MicroTAS 2011), 2011, p. 1176-1178Conference paper (Refereed)
    Abstract [en]

    Protein microarrays are useful tools for highly multiplexed determination of presence or levels of clinically relevant biomarkers in human tissues and biofluids. However, such tools have thus far been restricted to laboratory environments. Here, we present a novel 384-plexed easy to use lateral flow protein microarray device capable of sensitive (<50ng/ml) determination of antigen specific antibodies in less than ten minutes total assay time. Results were developed with gold nanobeads and could be recorded by a cell-phone camera or table top scanner. Excellent accuracy (AUC=99.4%) was achieved in comparison with an established glass microarray assay for 26 antigen-specific antibodies.

  • 45.
    Gantelius, Jesper
    et al.
    KTH, School of Biotechnology (BIO), Nano Biotechnology (closed 20130101).
    Bass, Tarek
    KTH, School of Biotechnology (BIO), Molecular Biotechnology (closed 20130101).
    Sjöberg, Ronald
    KTH, School of Biotechnology (BIO), Proteomics (closed 20130101). KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Nilsson, Peter
    KTH, School of Biotechnology (BIO), Proteomics (closed 20130101). KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Andersson-Svahn, Helene
    KTH, School of Biotechnology (BIO), Nano Biotechnology (closed 20130101).
    A Lateral Flow Protein Microarray for Rapid and Sensitive Antibody Assays2011In: International Journal of Molecular Sciences, ISSN 1661-6596, Vol. 12, no 11, p. 7748-7759Article in journal (Refereed)
    Abstract [en]

    Protein microarrays are useful tools for highly multiplexed determination of presence or levels of clinically relevant biomarkers in human tissues and biofluids. However, such tools have thus far been restricted to laboratory environments. Here, we present a novel 384-plexed easy to use lateral flow protein microarray device capable of sensitive (<30 ng/mL) determination of antigen-specific antibodies in ten minutes of total assay time. Results were developed with gold nanobeads and could be recorded by a cell-phone camera or table top scanner. Excellent accuracy with an area under curve (AUC of 98% was achieved in comparison with an established glass microarray assay for 26 antigen-specific antibodies. We propose that the presented framework could find use in convenient and cost-efficient quality control of antibody production, as well as in providing a platform for multiplexed affinity-based assays in low-resource or mobile settings.

  • 46.
    Gantelius, Jesper
    et al.
    KTH, School of Biotechnology (BIO), Nano Biotechnology.
    Hartmann, Michael
    KTH, School of Chemical Science and Engineering (CHE), Chemistry, Analytical Chemistry.
    Schwenk, Jochen M.
    KTH, School of Biotechnology (BIO), Proteomics.
    Roeraade, Johan
    Andersson-Svahn, Helene
    KTH, School of Biotechnology (BIO), Nano Biotechnology.
    Joos, Thomas O
    Magnetic bead-based detection of autoimmune responses using protein microarrays.2009In: New biotechnology, ISSN 1871-6784, Vol. 26, p. 269-276Article in journal (Refereed)
    Abstract [en]

    In the present study, a magnetic bead-based detection approach for protein microarrays is described as an alternative approach to the commonly used fluorescence-based detection system. Using the bead-based detection approach with applied magnetic force, it was possible to perform the detection step more rapidly as a result of the accelerated binding between the captured analyte in the microspot and the detection antibody, which was coupled to the magnetic beads. The resulting strong opacity shift on the microspots could be recorded with an ordinary flatbed scanner. In the context of autoimmunity, a set of 24 serum samples was analyzed for the presence of antibodies against 12 autoantigens using standard fluorescence and magnetic bead-based detection methods. Dynamic range, sensitivity, and specificity were determined for both detection methods. We propose from our findings that the magnetic bead-based detection option provides a simplified and cost effective readout method for protein microarrays.

  • 47.
    Gantelius, Jesper
    et al.
    KTH, School of Biotechnology (BIO), Nano Biotechnology (closed 20130101).
    Härlin, A.
    Elfversson, G.
    Nystrand, M.
    Andersson-Svahn, Helene
    KTH, School of Biotechnology (BIO), Nano Biotechnology (closed 20130101).
    Improved sensitivity on an allergen lateral flow microarray by means of dendritic amplification2009In: Proceedings of Conference, MicroTAS 2009 - The 13th International Conference on Miniaturized Systems for Chemistry and Life Sciences, Chemical and Biological Microsystems Society , 2009, p. 1136-1137Conference paper (Refereed)
    Abstract [en]

    Recently, paper-based substrates have been proposed as an alternative to commonly used activated glass slides for microarray patterning[1], used in conjunction with capillary driven lateral flow of sample and detection reagents through the membrane. While fluorescent detection reagents may be employed to achieve high sensitivity, gold nanoparticles can also be used to allow readout by means of common table top scanners or digital cameras. Here, we demonstrate first results from employing a dendritic, or layer-by-layer, amplification approach for a high-density lateral flow allergen protein microarray, indicating that substantially increased sensitivity can be achieved with very modest increase of assay handling and time requirements.

  • 48.
    Gantelius, Jesper
    et al.
    KTH, School of Biotechnology (BIO), Nano Biotechnology.
    Nystrand, M.
    Harlin, A.
    Elfverson, G.
    Schwenk, Jochen M.
    KTH, School of Biotechnology (BIO), Proteomics.
    Uhlén, Mattias
    KTH, School of Biotechnology (BIO), Proteomics.
    Eriksson-Karlström, Amelie
    KTH, School of Biotechnology (BIO).
    Andersson-Svahn, Helene
    KTH, School of Biotechnology (BIO), Nano Biotechnology.
    Evaluation of a lateral flow microarray assay systemArticle in journal (Other academic)
  • 49.
    Gantelius, Jesper
    et al.
    KTH, School of Biotechnology (BIO), Nano Biotechnology.
    Schwenk, Jochen M.
    KTH, School of Biotechnology (BIO), Proteomics.
    Hamsten, Carl
    KTH, School of Biotechnology (BIO).
    Neiman, Maja
    KTH, School of Biotechnology (BIO), Proteomics.
    Persson, Anja
    KTH, School of Biotechnology (BIO), Proteomics.
    Andersson-Svahn, Helene
    KTH, School of Biotechnology (BIO).
    A lateral flow protein microarray for rapid determination of contagious bovine pleuropneumonia status in bovine serum2010In: Journal of Microbiological Methods, ISSN 0167-7012, E-ISSN 1872-8359, Vol. 82, no 1, p. 11-18Article in journal (Refereed)
    Abstract [en]

    Novel analytical methods for a next generation of diagnostic devices combine attributes from sensitive, accurate, fast, simple and multiplexed analysis methods. Here, we describe a possible contribution to these by the application of a lateral flow microarray where a panel of recombinant protein antigens was used to differentiate bovine serum samples in the context of the lung disease contagious bovine pleuropneumonia (CBPP). Lateral flow arrays were produced by attaching nitrocellulose onto microscopic slides and spotting of the recombinant proteins onto the membranes. The developed assay included evaluations of substrate matrix and detection reagents to allow for short assay times and convenient read-out options, and to yield a total assay time from sample application to data acquisition of less than ten minutes. It was found that healthy and disease-affected animals could be discriminated (AUC = 97%), and we suggest that the use of an antigen panel in combination with the lateral flow device offers an emerging analytical tool towards a simplified but accurate on-site diagnosis.

  • 50. Griss, P.
    et al.
    Andersson, Helene
    KTH, Superseded Departments, Biotechnology.
    Stemme, Göran
    KTH, Superseded Departments, Signals, Sensors and Systems.
    Expandable microspheres for the handling of liquids2002In: Lab on a Chip, ISSN 1473-0197, E-ISSN 1473-0189, Vol. 2, no 2, p. 117-120Article in journal (Refereed)
    Abstract [en]

    Two novel concepts for controlled handling of liquids in microfluidic systems are presented: a one-shot micropump and a normally open one-shot valve based on thermo-expanding Expancel microspheres. Expancel microspheres are small spherical plastic particles that, when heated, increase their volume considerably. We show that liquid volumes in the nanoliter range can be actuated against a counter pressure of at least 100 kPa and fluid flow can be inhibited in a microchannel against pressures of at least 100 kPa.

123 1 - 50 of 137
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