Multiplexed fluorescent imaging which is essential for finding out how various biomolecules are spatially distributed in cells or tissues is disclosed. The present disclosure may obtain 10 or more different biomolecule images with one labeling and imaging by newly designing selection of fluorophores, detection spectral ranges, and signal unmixing algorithm. The present disclosure is a blind unmixing technology for unmixing an image without an emission spectrum of fluorophore, and in this technology, 4 pairs of fluorophores are used, and each pair consists of two fluorophores in which emission spectra are overlapped. Each pair of fluorophores is strongly excited by only one excitation laser. Two images with different detection spectral ranges are obtained for each pair, and two images are unmixed via mutual information minimization without fluorophore emission spectrum information. Two images also may be unmixed via Gram-Schmidt orthogonalization and fluorescence measurement based unmixing. This signal unmixing is repeated for each pair of fluorophores. Furthermore, a total of 10 or more fluorophores may be simultaneously used by adding two large stoke's shift fluorophores emitting light in wavelength ranges that does not overlap with the emission spectra of the above 8 fluorophores.

Methods of deep ultraviolet fluorescence (DUVF) and multi spectral imaging (MSI) detection are disclosed herein for the detection and identification of pathogens on plants. DUV light and visible or near-infrared light are used to illuminate plants or plant leaves such that the light intensity reflected or emitted by the plant or plant leaves can be used to identify the type of pathogen and measure the amount of pathogen on the plant or plant leaves and, additionally, be used to measure the plant's stress response to such pathogen. Also provided herein is a biophotonic analyzer device that uses both DUVF and MSI detection for the monitoring and surveillance of plant health and for the identification and enumeration of pathogens on plants or plant leaves.

An apparatus for analyzing a plant specimen is disclosed which includes a housing assembly adapted to be in i) an open configuration adapted to receive a plant specimen, and ii) a closed configuration wherein ambient light is controlled therein, a light source disposed in or coupled to the housing assembly, the light source adapted to shine light onto or through the plant specimen when the housing assembly is in the closed configuration, and a camera assembly coupled to the housing assembly, the camera assembly having an image sensor adapted to receive light from the plant specimen in i) a transmittance mode where light transmits through the plant specimen, or ii) a reflectance mode where light is reflected from the plant specimen, the image sensor adapted to thereby capture hyperspectral images of the plant specimen.

Disclosed is an apparatus and method for fluorescence excitation and detection. The apparatus comprises one or more light sources for providing excitation light for fluorescence excitation at an observation spot along an optical axis for excitation, an optical collection element for collecting fluorescence light generated by the excitation light at two or more different observation spots into two or more different measurement channels with an optical axis for collecting non-parallel to the optical axis for excitation of each of the one or more light sources, and, for each of the two or more measurement channels and thereby for each of the two or more observation spots, a dedicated optical detector for detecting fluorescence from the fluorescence light collected by the optical collection element.

A system for urine sample analysis, the system may include one or more transmitters for transmitting radiation; one or more sensors that are configured to receive received radiation that passed through the urine sample and to generate detection signals indicative of an intensity of the received radiation at multiple frequencies; detaching elements that are configured to detach the one or more transmitters and the one or more sensors to a toilet bowl; and a processor that is configured to participate in the urine sample analysis for determining a content of the urine sample based on the detection signals.

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.

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.

A fluorescence observation apparatus (1) includes an irradiation unit (10) that applies a plurality of kinds of excitation light (L11, L21) of mutually different wavelengths to a plurality of spatially or temporally different positions in a biological sample (5) that is labeled with a composite phosphor containing two or more kinds of fluorescent molecules (A, B) at a predetermined composition ratio, a detection unit (20) that detects fluorescence (L12, L22) generated at each of the plurality of positions by application of the irradiation unit (10), and a calculation unit (33) that determines a distribution of pieces of the composite phosphor on the basis of a fluorescence signal (S1, S2) that is obtained from a detection result of the detection unit (20) and that shows a fluorescence intensity corresponding to a position in the biological sample (5) of each piece of the fluorescence (L12, L22).

Integrated target waveguide devices and optical analytical systems comprising such devices are provided. The target devices include an optical coupler that is optically coupled to an integrated waveguide and that is configured to receive optical input from an optical source through free space, particularly through a low numerical aperture interface. The devices and systems are useful in the analysis of highly multiplexed optical reactions in large numbers at high densities, including biochemical reactions, such as nucleic acid sequencing reactions. The devices provide for the efficient and reliable coupling of optical excitation energy from an optical source to the optical reactions. Optical signals emitted from the reactions can thus be measured with high sensitivity and discrimination. The devices and systems are well suited for miniaturization and high throughput.

A method for scarless genome editing is disclosed. In particular, the method provides scarless genome modification by using homology directed repair (HDR) steps to genetically modify cells and remove unwanted sequences. This method can be used for genome editing, including introducing mutations, deletions, or insertions at any position in the genome without leaving silent mutations, selection marker sequences, or other additional undesired sequences in the genome.