According to exemplary embodiments of the present disclosure, it is possible to provide method, system, arrangement, computer-accessible medium and device to stimulate individual neurons in brain slices in any arbitrary spatio-temporal pattern, using two-photon uncaging of photo-sensitive compounds such as MNI-glutamate and/or RuBi-Glutamate with beam multiplexing. Such exemplary method and device can have single-cell and three-dimensional precision. For example, by sequentially stimulating up to a thousand potential presynaptic neurons, it is possible to generate detailed functional maps of inputs to a cell. In addition, it is possible to combine this exemplary approach with two-photon calcium imaging in an all-optical method to image and manipulate circuit activity. Further exemplary embodiments of the present disclosure can include a light-weight, compact portable device providing for uses in a wide variety of applications.

This disclosure includes an imaging system that is configured to image in parallel multiple focal planes in a sample uniquely onto its corresponding detector while simultaneously reducing blur on adjacent image planes. For example, the focal planes can be staggered such that fluorescence detected by a detector for one of the focal planes is not detected, or is detected with significantly reduced intensity, by a detector for another focal plane. This enables the imaging system to increase the volumetric image acquisition rate without requiring a stronger fluorescence signal. Additionally or alternatively, the imaging system may be operated at a slower volumetric image acquisition rate (e.g., that of a conventional microscope) while providing longer exposure times with lower excitation power. This may reduce or delay photo-bleaching (e.g., a photochemical alteration of the dye that causes it to no longer be able to fluoresce), thereby extending the useful life of the sample.

Disclosed herein are systems for imaging and ablating a sample. An imaging/ablating device (110) includes an optical assembly (112), a sample stage (114), and a receiver (116). The optical assembly (112) is disposed in an inverted position below the sample stage (114) and the receiver (116) is positioned above the sample stage (112). The optical assembly enables imaging of a sample disposed on the sample stage (114). The optical assembly (112) also enables ablation of a region of interest within the sample. The laser light propagated from the optical assembly during ablation propagates substantially in the same direction as the direction of travel of the ablation plume (20) toward the receiver (116).

A photon peak event detection system accepts an analog output from a photon sensor, directly digitizes the analogy output and includes a graphics processing unit (GPU) programmed to conduct a photon peak event detection in real-time via a photon count program that analyzes the digitized photon sensor output in sampling periods each having at least three consecutive data points to determine a local maximum among the consecutive data points and compare the local maximum to one or more predetermined thresholds to determine whether or not a photon was received in each sampling period, the algorithm providing photon counts to a phasor analysis program in the GPU. The phasor analysis program calculates pixelwise fluorescence lifetime phasor data in real-time and sends the data to a central processing unit.

An array of more than one digital micro-camera, along with the use of patterned illumination and a digital post-processing operation, jointly create a multi-camera patterned illumination (MCPI) microscope. Each micro-camera includes its own unique lens system and detector. The field-over-view of each micro-camera unit at least partially overlaps with the field-of-view of one or more other micro-camera units within the array. The entire field-of-view of a sample of interest is imaged by the entire array of micro-cameras in a single snapshot. In addition, the MCPI system uses patterned optical illumination to improve its effective resolution. The MCPI system captures one or more images as the patterned optical illumination changes its distribution across space and/or angle at the sample. Then, the MCPI system digitally combines the acquired image sequence using a unique post-processing algorithm.

Embodiments of the current disclosure are directed to systems, methods and apparatus for evaluating single cell secretion profiles. In some embodiments, the apparatus may be configured to analyze substances expressed by a biological cell and may include a first compressible substrate, and a second substrate configured for removable sealing attachment with the first substrate. In some embodiments, upon attachment of the second substrate with the first substrate, an assembly is formed such that the open side of the plurality of chambers are covered by the second substrate, and a portion of each of the plurality of capture areas are exposed in each of the chambers.

An optical system is presented for optically imaging a sample including a nanoscale object. The optical system includes an imaging lens, an illumination source configured to provide an excitation light, a detector and a substrate for supporting the sample. A sample interface, arranged to reflect the excitation light, is formed between the sample and a first side of the substrate facing the sample when the sample is applied on the substrate. The optical imaging system is arranged such that the excitation light is sent into the substrate via the imaging lens and such that the detector receives a reference light and a scattered light. The reference light comprises a part of the excitation light reflected at the sample interface and collected by the imaging lens and the scattered light comprises a part of the excitation light scattered by the nanoscale object and collected by the imaging lens. The optical system is configured such that the nanoscale object is imaged at the detector, in response to the excitation light, by an optical contrast of an interference pattern between the reference light and the scattered light. The substrate comprises an optical coating disposed on the first side of the substrate such that the sample is in contact with the optical coating when the sample is applied on the substrate. A degree of reflection of the excitation light at the sample interface is such that the optical contrast is larger compared to the optical contrast obtained with the sample interface formed without the optical coating.

An optical imaging system includes a light source, a light detector and an aperture plate. The light source includes a plurality of light emitting devices which emit light that is directed toward an object to be illuminated. The light detector is positioned to view the object illuminated by the light source. The aperture plate is positioned relative to the light source to block a first portion of the light emitted by the light source and to allow a second portion of the light emitted by the light source to pass therethrough to illuminate a pre-defined area of the object. The aperture plate includes a plurality of spaced apart apertures formed through the thickness thereof. Each aperture corresponds to a respective light emitting device. Each aperture of the aperture plate is defined by a first opening formed in the thickness of the aperture plate and a second opening formed in the thickness of the aperture plate. The second opening partially overlaps the first opening and is partially offset from the first opening. The first and second openings' planar shapes match the shape of the desired illumination area, with the first openings being smaller than the second openings. A method for illuminating a defined area of an object includes the steps of energizing one or more light emitting devices of a light source in an optical imaging system, which energized light emitting device or devices emit light that is directed toward the object to be illuminated. The light is passed through particularly-shaped apertures, such as described above, formed in an aperture plate positioned between the light source and the object to be illuminated. The apertures in the plate only allow light passing therethrough to impinge on the object at a pre-defined area thereof.

A prediction algorithm determines synthetic fluorescence images on the basis of measurement images. A validation of the synthetic fluorescence images can be effected on the basis of reference images which are captured after the measurement images or are captured for a separate sample. Alternatively or additionally, a training of the prediction algorithm can be effected on the basis of training images which are captured after the measurement images or are captured for a separate sample.

Provided herein are systems and methods for imaging using a microscope system comprising removeable or replaceable component parts. Such systems and methods employ semi-kinetic coupling for easy, tool-free attachment of the microscope system to a baseplate. Systems and methods provided herein may comprise simultaneous imaging and stimulation using a microscope system. The microscope system can have a relatively small size compared to an average microscope system.