Fiducial markers are provided on patterned arrays of the type that may be used for molecular analysis, such as sequencing. The fiducials may have configurations that enhance their detection in image or detection data, that facilitate or improve processing, that provide encoding of useful information, and so forth. Examples of the fiducials may include a non-closed shape that may encode information, allow for bubbles to escape during manufacture, and provide additional advantages over closed shape fiducials.

The present invention relates to an illumination filter system (2) for medical imaging, in particular multispectral fluorescence imaging, as performed e.g. in a microscope (1) or endoscope, such as a surgical microscope, in particular a surgical multispectral fluorescence microscope, comprising a first optical filter (35). The present invention also relates to an observation system (3) for medical imaging, in particular multispectral fluorescence imaging, as performed e.g. in a microscope (1) or endoscope, in particular a multispectral fluorescence microscope, comprising a beam splitter (21) adapted to split a light image (13) into a first light portion (16, 17) along a first light path (18) and a second light portion (20) along a second light path (19). To improve known illumination filter systems and observation systems, so these systems work with one light source only, are capable of capturing simultaneously at least one fluorescence signal and a signal of visible reflected light and allow a homogeneous illumination for obtaining different images from the object illuminated, the first optical filter (36) is adapted to quench light of at least one fluorescence excitation band within the visible spectrum in the illumination filter system (2) of the present invention, and the first light portion (16, 17) comprises at least one fluorescence emission band (Em.1, Em.2) in the visible spectrum and the second light portion (20) portion comprises a visible reflected light (VISR) in the observation system (3) of the present invention.

The invention relates to a localization microscopy method for localizing signal sources. Here, at least once for each pixel of a detector, values of an error parameter are ascertained and stored in a calibration data record in a manner assigned to the relevant pixel. Captured image data are used to identify regions of origin of signal sources and fit a point spread function to the pixel values of the respective regions of origin. The respective signal source is localized on the basis of the point spread function. The pixel-specific error parameter of each pixel can be compared to a threshold. If the threshold is exceeded, these pixels are either ignored or replaced by means of interpolation when fitting the point spread function. In addition or as an alternative thereto, the real noise performance of the pixels is ascertained and corrected on the basis of derived pixel-specific error parameters.

To minimize cross talk in systems and methods for detecting two or more different optical signals emitted from each of a plurality of reaction receptacles, an excitation signal associated with each of the optical signals has a known excitation frequency, and any detected signal having a frequency that is inconsistent with the excitation frequency is discarded. The receptacles are moved relative to optical sensors configured to detect each unique optical signal from an associated receptacle, and to further minimize cross talk, the optical sensors are arranged so that only one reaction receptacle at a time is in a signal detecting position with respect to one of its associated optical sensors, and the optical sensors are grouped by the optical signal they are configured to detect so that a first optical signal is detected from each of the reaction receptacles before a second optical signal is detected from the reaction receptacles.

A method of processing a sample in a receptacle comprising a plurality of chambers. Each of the chambers is connected to at least one other chamber by a portal and at least a first one of the chambers is formed of a flexible material. The method includes the steps of causing gas bubbles contained in the first chamber to accumulate in a portion of the first chamber, applying a compressive external force to the first chamber to cause some or all of the liquid contents of the first chamber to flow into an interconnected second chamber through a portal connecting the first and second chambers; and preventing the gas bubbles accumulated in a portion of the first chamber from flowing through the portal into the second chamber.

The present invention relates to a method of identifying and/or distinguishing materials by means of luminescence, wherein at least one luminescent substance is incorporated into the material and/or applied onto the material and the luminescence behaviour of the substance is analysed after excitation by means of radiation, and the use thereof for identifying and/or sorting and/or recycling and/or authenticating and/or performing a quality check and/or formulation check on materials.

The present disclosure relates to spatially modulating the light source used in microscopy. In some cases, a light source projects a sequence of two-dimensional spatial patterns onto a sample using a spatial light modulator. In some cases, the spatial patterns are based on Hadamard matrices. In some cases, an imaging device captures frames of image data in response to light emitted by the sample and orthogonal components of the image data are analyzed by cross-correlating the image data with the spatial pattern associated with each frame. A microscope may be calibrated by illuminating a sample with the sequence of spatial patterns, capturing image data, and storing calibration that maps each pixel of the spatial light modulator to at least one pixel of the imaging device.

An apparatus and method are provided for differentiating multiple detectable signals by excitation wavelength. The apparatus can include a light source that can emit respective excitation light wavelengths or wavelength ranges towards a sample in a sample retaining region, for example, in a well. The sample can contain two or more detectable markers, for example, fluorescent dyes, each of which can be capable of generating increased detectable emissions when excited in the presence of a target component. The detectable markers can have excitation wavelength ranges and/or emission wavelength ranges that overlap with the ranges of the other detectable markers. A detector can be arranged for detecting an emission wavelength or wavelength range emitted from a first marker within the overlapping wavelength range of at least one of the other markers.

Fiducial markers are provided on a patterned array of the type that may be used for molecular analysis, such as sequencing. The fiducial markers may have configurations and layouts that enhance their detection in image or detection data, that facilitate or improve processing, that provide encoding of useful information, and so forth. Examples of the fiducial markers may include non-rectilinear layouts that may provide for more robust location of both the fiducial markers and sites of the patterned array.

Methods and systems are disclosed for extracting an image of a target fluorophore in a biological material, which involve inducing both autofluorescence of the biological material and fluorescence of the fluorophore, acquiring an image arising from both the autofluorescence of the biological material and the fluorophore, and an image arising only from the autofluorescence, subtracting the two images to produce an image representing only the fluorophore, wherein relative intensities of the excitation light used to induce the autofluorescence and the fluorescence are modulated prior to acquiring the images.