Abstract: Microscopy imaging that allow for multiple mRNAs, proteins and metabolites to be spatially resolved at a subcellular level provides valuable molecular information which is a crucial factor for understanding tissue heterogeneity as for example within the tumor micro environment. The current invention describes a method (SUMI-Seq) which combines the use of Spatial Unique Molecular Identifier in situ sequencing and in vitro sequencing of rolonies derived from rolling circle amplification from padlock oligonucleotides targeting portion of RNA or cDNA transcript at a subcellular level with less limitation in the amount of transcripts and the length of the sequence that can be analyzed. Apart from padlocks oligonucleotides, the SUMI-Seq method can also be applied using circular oligonucleotides to spatially resolve proteins and metabolites to provide multiomics results.

Abstract: The invention relates to a system, comprising: a) a sample processing unit, comprising an input port and an output port coupled to a rotating container having at least one sample chamber, the sample processing unit configured provide a first processing step to a sample or to rotate the container so as to apply a centrifugal force to a sample deposited in the chamber and separate at least a first component and a second component of the deposited sample; and b) a sample separation unit coupled to the output port of the sample processing unit, the cell separation unit comprising separation column holder (42), a pump (64) and a plurality of valves (1-11) configured to at least partially control fluid flow through a fluid circuitry and a separation column (40) positioned in the holder, the separation column configured to separate labeled and unlabeled components of sample flowed through the column.

Abstract: The present invention provides a method for generation, isolation, detection and/or analysis of cardiomyocytes derived from a starting cell composition comprising pluripotent and/or multipotent stem cells, the method comprising the steps a) differentiating said pluripotent and/or multipotent stem cells into cardiovascular cells, thereby generating a sample comprising cardiomyocytes and non-cardiomyocytes; and b) labeling the non-cardiomyocytes of said sample with more than one antibody or antigen binding fragment thereof specific for antigens of non-cardiomyocytes and c) depleting said labeled non-cardiomyocytes from said sample; and optionally d) labeling the cardiomyocytes of said sample with at least one antibody or antigen binding fragment thereof specific for antigen(s) of cardiomyocytes; and e) enriching said labeled cardiomyocytes and detecting and isolating cardiomyocyte subtypes derived from said pluripotent and/or multipotent stem cells.

Abstract: The invention is directed to a conjugate having the general formula (I) Xn-P-YmBo (I) with X is an detection moiety, P is a spacer unit, Y an antigen recognizing moiety, B an oligonucleotid comprising 2 to 100 nucleotide residues and n, m, o are independent integers between 1 and 100 wherein P and B are covalently bound to Y and X is covalently bound to P and wherein X is erasable. Further, the invention is directed to a library of such conjugates and a method of detecting target cells utilizing the conjugates or the library of conjugates.

Abstract: Systems and methods are described for analyzing a plurality of biological samples with a plurality of fluorescent reagents in an automated fashion. The system may include two optical systems, a fluorescence imaging system and an optical quenching system. These two systems may be placed laterally adjacent to one another, and generally beneath one of the wells of sample-containing microtiter plate. The biological sample contained therein may be stained with a series of fluorescent reagents, with the fluorescence quenched between stainings. By precise positioning of the sample with respect to the imaging system, the sample may be imaged with a plurality of serially applied reagents.

Abstract: A cell modification device, comprising a centrifugation chamber with at least one cell modifying surface with a normal vector having an angle of 135-45° to the rotational axis of the centrifugation chamber, wherein the centrifugation chamber comprises at least one input/output port and the cells to be modified are immobilized at the cell modifying surfaces by the rotation of the centrifugation chamber at 2 to 2000 g. In an embodiment, the device is used as a point-of-care and/or portable device. Further, the present disclosure describes software that, when executed by a processor, causes the device to perform the disclosed functions.

Abstract: The present invention provides a differentiation medium for differentiation and expansion of a multicellular aggregation in suspension derived from human pluripotent stem cells that has been induced to differentiate to an artificial tissue structure such as artificial neural tissue, said medium comprising a basal medium for animal or human cells, wherein said differentiation medium has a viscosity between 1.7 mPa*s and 1500 mPa*s. Said viscosity is achieved by the presence of a viscosity enhancer such as methyl cellulose, carboxymethyl cellulose, or hydroxy ethyl cellulose in said differentiation medium. Also disclosed are an in-vitro method for obtaining artificial neural tissue and a kit comprising said differentiation medium.

Abstract: The invention provides a system that comprises pharmaceutical agents for use in immunotherapy for reducing the side-effects of an antigen-recognizing receptor against antigen-expressing non-target cells in an individual. The system includes an antigen-recognizing receptor that specifically recognizes an antigen on target cells and at least on one hematopoietic cell type in the individual. The antigen-recognizing receptor is exemplified by chimeric antigen receptors (CAR) be expressed on the surface of an immune effector cells. The system also includes hematopoietic cells resistant to recognition of the same antigen by the antigen-recognizing receptor.

Abstract: Microscopy imaging that allow for multiple mRNAs to be resolved at a single cell level provides valuable information regarding transcript amount and localization, which is a crucial factor for understanding tissue heterogeneity, the molecular development and treatment of diseases. The current invention describes a method (Fly FISH) which combined the use of padlock oligonucleotides as fluorescence in situ hybridization (FISH) probes for detection and sequencing targeted portion of RNA or cDNA transcript at a cellular level with less limitation in the amount of transcripts and the length of the sequence that can be analyzed. Padlocks probes containing various barcodes in their core are utilized both as FISH probes and also to capture RNA portion that can be sequenced. The same barcodes can be used to selectively prime a rolling circle amplification and amplify a subset of transcripts coming from a specific region that have been tagged as of interest during the probing steps.

Abstract: The invention is directed to a method for detecting a target moiety in a sample of biological specimens by providing a conjugate with the general formula (I) characterized in contacting the sample of biological specimens with at least one conjugate (I), thereby labeling the target moiety recognized by an antigen recognizing moiety with the conjugate (I); exciting the labelled target moieties with light having a wavelength within the absorbance spectrum of the fluorescent moiety FL; detecting the labelled target moieties by detecting the fluorescence radiation emitted by the fluorescent moiety FL and degrading the fluorescent moiety FL of the labelled target moieties by irradiating the conjugate with light having a wavelength within the absorbance spectrum of fluorescent moiety FL for a time sufficient to deliver enough energy to reduce the fluorescence radiation emitted by the fluorescent moiety FL at least by 75% of the initial fluorescence radiation.

Abstract: The invention is directed to a method for preparing a detection reagent for CAR expressing cells according to general formula (I) (X-S)m-D ??(I) wherein D is a detection moiety, S a spacer unit and X a polypeptide comprising 10 to 80 amino acids and a N3 group, which binds to the antigen binding domain of a chimeric antigen receptor of a cell and m=4-200, characterized in reacting p detection moieties D with a spacer S according to general formula (II) wherein R is a leaving group, q=1 to 5, and n=4 to 200 and p is 4-250 thereby forming modified detection moiety Sp-D and reacting the modified detection moiety Sp-D with a plurality of polypeptides X to yield (X-S)m-D.

Abstract: A multilevel, disposable cartridge may have a plurality of fluid wells or reservoirs holding a plurality of reagents, and a sample viewing area to view a biological sample. The reagents may be applied sequentially to the sample by opening a specific valve formed in an elastomeric, flexible layer under the fluid well. The fluid channels may be formed in a first rigid plastic layer to conduct the reagent to the sample. Pneumatic channels for applying suction or pressure to the valves may be formed in a second rigid, plastic layer.

Abstract: The present invention provides the use of a packaging cell line for the production of VSV-G pseudotyped retroviral vector particles or virus like particles thereof, wherein said packaging cell line is negative for Low-Density Lipoprotein Receptor (LDLR), optionally said packaging cell line stably expresses VSV-G. A method for producing said VSV-G pseudotyped retroviral vector particles or virus like particles thereof is disclosed as well as said particles obtained by said method.

Abstract: The invention is directed to the use or a method of using of a cell expressing a chimeric antigen recognition structure (CAR) bound to an adapter molecule, wherein: i) the CAR structure comprises a) an antigen binding domain specific for the tag of the adapter molecule, b) a transmembrane domain c) an intracellular domain wherein the ii) the adapter molecule comprises a) a tag which is specific for the antigen binding domain of the CAR structure b) a polypeptide comprising an antigen recognizing domain specific for an antigen expressed on the target cell, wherein the polypeptide comprises a heavy chain and a light chain connected by a sulphur atom-linker-sulphur atom bridge and wherein the linker is covalently bound to a single tag for targeting target cells, especially human tumor cells.

Abstract: The invention is directed to a method for detecting a target moiety in a sample of biological specimens by: a) providing at least one conjugate with the general formula (I) Xn—P—Ym, b) contacting the sample of biological specimens with at least one conjugate, thereby labeling the target moiety recognized by the antigen recognizing moiety Y with the conjugate c) exciting the labelled target moieties with light having a wavelength within the absorbance spectrum of fluorescent moiety X d) detecting the labelled target moieties by detecting the fluorescence radiation and e) degrading the fluorescent moiety X of the labelled target moieties by irradiating the conjugate with light having a wavelength within the absorbance spectrum of fluorescent moiety X Use of the method in fluorescence microscopy, flow cytometer, spectrofluorometry, cell separation, pathology or histology.

Abstract: The invention is directed to disposable cartridge for electroporation of cells, comprising a fluid compartment in an interior of the disposable cartridge; a first fluid port for providing cell suspension to the fluid compartment, and a second fluid port for delivering a fluid comprising at least one compound to be electroporated into the cells to the fluid compartment; a first electrode and a second electrode disposed in the fluid compartment; at least one exit port which delivers the fluid from the fluid compartment wherein the first and second fluid port have a fluid communication to a mixing channel which has a fluid communication to the fluid compartment, and a third ground electrode and a fourth ground electrode.

Abstract: The present invention provides a composition comprising i) a pseudotyped retroviral vector particle or virus-like particle thereof comprising a) one envelope protein with antigen-binding activity, wherein said envelope protein is a recombinant protein that does not interact with at least one of its native receptor(s) and is fused at its ectodomain to a polypeptide comprising an antigen binding domain specific for a tag of a tagged polypeptide, and wherein said envelope protein is protein G, HN or H derived from the Paramyxoviridae family, and b) one envelope protein with fusion activity derived from the Paramyxoviridae family, and ii) said tagged polypeptide, wherein said tagged polypeptide binds specifically to an antigen expressed on the surface of a target cell, thereby transducing the target cell with said retroviral vector particle or thereby inducing uptake of the virus-like particle into the target cell.

Abstract: There is provided a microscope device comprising: a microscope objective (18), a sample stage (12) for holding a sample (16) and moving the sample in a plane perpendicular to the optical axis of the microscope objective, a light source (20) and an illumination pattern generator unit (22) for illuminating the sample with a line-type illumination pattern, an area-detector detector (40, 44) for detecting light from the sample collected by the microscope objective, a scanning unit (24) for moving the illumination pattern in a scanning direction across the field of view, wherein the scanning direction is perpendicular to the longitudinal axis of the illumination pattern, and a control unit (42) configured to selectively operate the microscope arrangement in a normal magnification mode or in a lower magnification mode, wherein in the lower magnification mode the sample is moved in a direction antiparallel to the scanning direction while the illumination pattern is moved across the field of view by the scanning unit

Abstract: The invention provides a system that comprises pharmaceutical agents for use in immunotherapy for reducing the side-effects of an antigen-recognizing receptor against antigen-expressing non-target cells in an individual. The system includes an antigen-recognizing receptor that specifically recognizes an antigen on target cells and at least on one hematopoietic cell type in the individual. The antigen-recognizing receptor is exemplified by chimeric antigen receptors (CAR) be expressed on the surface of an immune effector cells. The system also includes hematopoietic cells resistant to recognition of the same antigen by the antigen-recognizing receptor.

Abstract: There is provided a microscope device comprising: an objective (1); an actuator (6) for adjustment of a distance between the objective and a sample (3) in a focusing direction z during a focusing action to reach and hold a desired focus position, an absolute z-position detector (7) for measuring a z-position of the actuator in the focusing direction, an autofocus light source (11); a first optical arrangement (12, 13) for generating a focused spot (31) of autofocus light from the light source in the backfocal plane (14) of the objective (1) at a position offset from the optical axis of the objective by a lateral offset distance (of), such that the objective generates a collimated incoming beam (22) of autofocus light, which is directed at an oblique angle (13) relative to the optical axis (15) of the objective onto the substrate; and a second optical arrangement (13, 16) for generating collimated outgoing beams (34, 35) of autofocus light from the collimated incoming beam as reflected by a substrate surface (