Apparatus and methods for fluorescence imaging using radiofrequency multiplexed excitation. One apparatus splits an excitation laser beam into two arms of a Mach-Zehnder interferometer. The light in the first beam is frequency shifted by an acousto-optic deflector, which is driven by a phase-engineered radiofrequency comb designed to minimize peak-to-average power ratio. This RF comb generates multiple deflected optical beams possessing a range of output angles and frequency shifts. The second beam is shifted in frequency using an acousto-optic frequency shifter. After combining at a second beam splitter, the two beams are focused to a line on the sample using a conventional laser scanning microscope lens system. The acousto-optic deflectors frequency-encode the simultaneous excitation of an entire row of pixels, which enables detection and de-multiplexing of fluorescence images using a single photomultiplier tube and digital phase-coherent signal recovery techniques.

Provided is a novel fluorescent probe.

A compound of the following general formula (I) or a salt thereof.

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A system and method for depth-resolved imaging of fluorophore concentrations in tissue uses a pulsed light source stimulus wavelength to illuminate the tissue; and a time-gated electronic camera such as a single-photon avalanche detector camera to observe the tissue in multiple time windows after start of each light pulse. A filter-changer or tunable filter is between the tissue and the electronic camera with fluorescent imaging settings and a stimulus wavelength setting, and an image processor receives reflectance images and fluorescent emissions images from the time-gated camera and processes these images into depth and quantity resolved images of fluorophore concentrations in the tissue.

Systems, devices, and methods for producing an optimized phase mask for use in a single-molecule orientation localization microscopy (SMOLM) imaging system are disclosed.

Double sided flow cell and other substrate imaging systems, such as imaging systems used in nucleic acid sequencing and similar processes. In one example, the imaging system includes a flipper to facilitate imaging different surfaces of the flow cell or other substrate. In another example, the imaging system includes two optical systems for imaging different surfaces of the flow cell or other substrate. In another example, the imaging system is an immersion system. In these and other examples, the system may include an auto-focus sub-system configured to accurately focus the optics on one surface of the double sided flow cell without interference from the other surface of the flow cell.

Provided is a fluorescence reader that uses two excitation channels and can read up to seven different fluorescent dyes in a single run. Each excitation channel has one light source and one single excitation filter and one dichroic mirror. One excitation channel is capable of exciting multiple fluorescent dyes and can be used to distinguish multiple dyes in combination with multiple emission filters. The excitation channels are driven by a motor that can automatically switch the two excitation channels for taking images of up to seven different fluorescent dyes. An algorithm to calibrate the crosstalk between different fluorescent dyes is also provided. Also provided is a method for analyzing digital PCR data using a ratio of two fluorescence emission readings.

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.

Provided is an information processing apparatus (100) including: an image acquiring unit (112) that acquires captured image information of a sample (20) dyed with a fluorescent dye reagent (10), an information acquiring unit (111) that acquires information related to the fluorescent dye reagent (10), a correcting unit (131) that corrects the luminance of the captured image information using a fluorescence fading coefficient that represents the rapidness at which the fluorescence intensity of the fluorescent dye reagent (10) drops, the fluorescence fading coefficient being included in the fluorescent dye reagent (10), and a calculating unit (132) that calculates information corresponding to fluorescent molecules in the captured image information, using the corrected luminance.

The invention provides novel non-invasive in vitro methods for assessing the metabolic condition of oocytes and/or embryos with fluorescence lifetime imaging microscope, that can be used, for example, in assessment of oocytes and embryos in assisted reproductive technologies.

A filter unit for a Raman microscope mounted with a dark-field objective lens unit includes a frame body, a plurality of UV-LED elements that is disposed around a window part of the frame body to emit UV light, and a long-pass filter that is supported to the frame body to cover the window part of the frame body and transmits a light having a wavelength longer than the wavelength of the UV light. The filter unit has a dark-field UV irradiation function, and is able to impart a fluorescence observation function to the Raman microscope.