The invention enables efficient, rapid, and sensitive enumeration of living cells by detecting microscopic colonies derived from in situ cell division using large area imaging. Microbial enumeration tests based on the invention address an important problem in clinical and industrial microbiology—the long time needed for detection in traditional tests—while retaining key advantages of the traditional methods based on microbial culture. Embodiments of the invention include non-destructive aseptic methods for detecting cellular microcolonies without labeling reagents. These methods allow for the generation of pure cultures which can be used for microbial identification and determination of antimicrobial resistance.

A fluorescence microscope, includes an optical system configured to collect fluorescent light emitted from different fluorophore species within a field of view and to focus the fluorescent light for detection, a spectral splitting arrangement configured to split the fluorescent light into at least two spectrally different fluorescent light components, a multi-channel detector system including at least two image sensors configured to detect at least two spatial light intensity distributions based on the at least two spectrally different fluorescent light components, each spatial light intensity distribution representing an image of the object over the field of view, and a processor configured to determine spatial distributions of the different fluorophore species based on a spectral unmixing analysis of each spatial light intensity distribution, wherein the processor is further configured to obtain compensation information and to determine a spatial distribution of each fluorophore species by taking into account the compensation information.

Embodiments disclosed include methods and apparatus for Fluorescent Enhanced Photothermal Infrared (FE-PTIR) spectroscopy and chemical imaging, which enables high sensitivity and high spatial resolution measurements of IR absorption with simultaneous confocal fluorescence imaging. In various embodiments, the FE-PTIR technique utilizes combined/simultaneous OPTIR and fluorescence imaging that provides significant improvements and benefits compared to previous work by simultaneous detection of both IR absorption and confocal fluorescence using the same optical detector at the same time.

The present disclosure features portable wide field fluorimeter systems, e.g., in the form of low-cost mobile platforms, and methods to perform fluorometric assays to detect a change in fluorescence intensity in liquid samples, e.g., caused by the presence of a target analyte, e.g., a protein, e.g., an enzyme (e.g., β-lactamase) expressed by a target pathogen in a liquid sample in a point-of-care setting. In some implementations, a portable system for detecting a change in fluorescence intensity in a liquid sample includes a microfluidic device, an optical assembly including an emission filter and one or more lenses, and an analyzer device that collects and processes a fluorescent signal for the detection of a target analyte produced by the target pathogen present in the liquid sample.

High-throughput hyperspectral imaging systems are provided. According to an aspect of the invention, a system includes an excitation light source; an objective that is configured to image excitation light onto the sample, such that the excitation light causes the sample to emit fluorescence light; a channel separator that is configured to separate the fluorescence light into a plurality of spatially dispersed spectral channels; and a sensor. The excitation light source includes a light source and a plurality of lenslet arrays. Each of the lenslet arrays is configured to receive light from the light source and to generate a pattern of light, and the patterns of light generated by the lenslet arrays are combined to form the excitation light. The objective is configured to simultaneously image each of the patterns of light to form a plurality of parallel lines or an array of circular spots at different depths of the sample.

The present disclosure is drawn to microfluidic devices. The microfluidic device includes a microfluidic well, a layered composite stack, and an optical sensor. The layered composite stack includes an optical filter composited with an etch-stopping layer. The optical filter defines the microfluidic well. The optical sensor is associated with the microfluidic well and has the optical filter positioned therebetween.

A fluorescence detection system, including apparatus and methods, suitable for qPCR and other fluorescence-based analyses. The system may comprise various components, including a stage, an illumination module, a detection module, and an optical relay structure. The stage may be configured to support a sample holder. The illumination module may include one or more discrete light sources configured to produce excitation light. The detection module may be configured to detect fluorescence emission light produced, in response to the excitation light, by a fluorescent sample positioned in the sample holder. The optical relay structure may include a beamsplitter assembly configured to direct the excitation light from the illumination module along an illumination path to the sample holder and to direct the fluorescence emission light from the sample holder along a response path to the imaging module. The system may enhance the quality of excitation light hitting samples in the sample holder.

Provided herein is an apparatus for assessing a fluorescence characteristic of a gemstone. The apparatus comprises an optically opaque platform for supporting a gemstone to be assessed, one or more light source to provide uniform UV and non-UV illumination, an image capturing component, and a telecentric lens positioned to provide fluorescent images of the illuminated gemstone to the image capturing component. Also provided are methods of fluorescence analysis based on images collected using such an apparatus.

A sample observation device includes an irradiation unit that irradiates a sample with planar light, a scanning unit that scans the sample in one direction with respect to an irradiation surface of the planar light, an image formation unit that forms images of fluorescent light and scattered light from the sample, an imaging unit that outputs first image data based on a light image of the fluorescent light and second image data based on a light image of the scattered light, an image processing unit that generates a fluorescent light image on the basis of a plurality of pieces of first image data and generates a scattered light image on the basis of a plurality of pieces of second image data, and an analysis unit that specifies an area in which there is the sample in the fluorescent light image on the basis of the scattered light image, and sets an analysis area in the fluorescent light image on the basis of the area in which there is the sample.

A biosensor is provided. The biosensor includes a substrate, photodiodes, pixelated filters, an excitation light rejection layer and an immobilization layer. The substrate has pixels. The photodiodes are disposed in the substrate and correspond to one of the pixels, respectively. The pixelated filters are disposed on the substrate. The excitation light rejection layer is disposed on the pixelated filter. The immobilization layer is disposed on the excitation light rejection layer.