Provided herein are methods and systems for the analysis of biomarkers, and methods of providing diagnoses and/or prognoses therewith. In particular, methods and systems for performing biomarker ratio imaging microscopy (BRIM) are provided, as well as methods of using BRIM for the analysis of biomarker pairs (e.g., CD44/CD24, N-cadherin/E-cadherin, CD74/CD59, etc.) diagnosis and/or prognosis of cancer (e.g., ductal carcinoma in situ).

A method for imaging tissue is provided. The method includes illuminating a target tissue via a light-emitting diode. The method further includes acquiring a plurality of timed images of the target tissue including an excitation image corresponding to an excitation state of the light-emitting. Additionally, the method includes generating a relative lifetime map of the target tissue based on the plurality of timed images.

In a sample observation device, an image acquisition unit 6 acquires a plurality of pieces of image data of a sample in a Y-axis direction, and an image generation unit generates luminance image data on luminance of the sample on the basis of the plurality of pieces of image data, binarizes luminance values of each of the plurality of pieces of image data to generate a plurality of pieces of binarized image data, and generates area image data on an existing area of the sample on the basis of the plurality of pieces of binarized image data.

A hyperspectral detection approach is used in combination with narrow linewidth illumination for fluorescence excitation. A more efficient fluorophores excitation and image capturing may be provided, and thus high-quality data for subsequent hyperspectral analysis may be obtained. An apparatus, a method, a system and a computer program are disclosed.

The technology disclosed present systems and methods to produce an enhanced resolution image from images of a target using structured illumination microscopy (SIM). The method includes transforming at least three images of the target captured by a sensor in a spatial domain into a Fourier domain to produce at least three frequency domain matrices that each include first blocks of complex coefficients and redundant second blocks of complex coefficients that are conjugates to the first blocks. The method includes reducing computing resources required to produce the enhanced resolution image by using first blocks of complex coefficients to produce at least three phase-separated half-matrices in the Fourier domain. The method includes performing one or more intermediate transformation on the phase-separated half-matrices to produce realigned shifted half-matrices. The method includes calculating complex coefficients of second blocks in the Fourier domain to produce full matrices from half-matrices.

Systems and methods are provided for Quantitative Phase Microscopes (QPM) having laser systems including one or more of laser scissors and laser tweezers. In one embodiment, the system includes one or more structural elements, such as a stage and dichroic plate for operation of a QPM with laser scissors/tweezers. Another embodiment is directed to a method of operating a QPM system having laser scissors/tweezers. One or more solutions are provided for biodmedical applications of a QPM system including simulation and analysis of trauma on cellular structures and organelles. Processes are also provided for simulation and analysis of traumatic injury, including imaging and analysis of astrocytes.

The present disclosure generally relates to a molecular construct for multiphoton fluorescence microscopy imaging. The molecular construct has a first, non-fluorescent configuration (2PAP-C) and a second, fluorescent configuration (2PAP-CL), and comprises a two-photon absorbing probe (2PAP) linked to a photochromic molecule that can be reversibly changed from a first colored isomeric form (C) to a second colorless isomeric form (CL). The first colored form (C) can be isomerized to the second colorless isomeric form (CL) upon absorption of two photons by the two-photon absorbing probe (2PAP). The present disclosure also relates to a method for analyzing a target structure in a multiphoton microscope utilizing the molecular construct. Furthermore, the present disclosure relates to an antibody tagged with the molecular construct, and to the use of the molecular construct for imaging a target structure.

The present invention, among other things, provides technologies for detecting and/or quantifying nucleic acids in cells, tissues, organs or organisms. In some embodiments, through sequential barcoding, the present invention provides methods for high-throughput profiling of a large number of targets, such as transcripts and/or DNA loci.

A device for luminescent imaging includes an array of imaging pixels, a photonic structure over the array of imaging pixels, and an array of features over the photonic structure. A first feature of the array of features is over a first pixel of the array of imaging pixels, and a second feature of the array of features is over the first pixel and spatially displaced from the first feature. A first luminophore is within or over the first feature, and a second luminophore is within or over the second feature. The device includes a radiation source to generate first photons having a first characteristic at a first time, and generate second photons having a second characteristic at a second time. The first pixel selectively receives luminescence emitted by the first and second luminophores responsive to the first photons at the first time and second photons at the second time, respectively.

A direct structured illumination microscopy (dSIM) reconstruction method is provided. First, a time domain modulation signal is extracted through a wavelet. Then, an incoherent signal is converted into a coherent signal. Next, an accumulation amount at each pixel is calculated. Finally, a super-resolution image is generated by using a correlation between signals at different spatial positions. An autocorrelation algorithm of dSIM is insensitive to an error of a reconstruction parameter. dSIM bypasses a complex frequency domain operation in structured illumination microscopy (SIM) image reconstruction, and prevents an artifact caused by the parameter error in the frequency domain operation. The dSIM algorithm has high adaptability and can be used in laboratory SIM, nonlinear SIM imaging systems, or commercial systems.