Abstract: Zinc oxide compositions as well as techniques for plasmonic enhancement of absorption in sunscreen applications are provided herein. A method includes selecting one or more metal particles to be used in conjunction with one or more zinc oxide particles in a sunscreen composition, wherein said selecting is based on the plasmon resonance frequency associated with each of the metal particles; and embedding the one or more selected metal particles into each of the one or more zinc oxide particles.

Abstract: A system or method for analyzing a sample include an input light source, an excitation light source positioned to receive light from the input light source and to sequentially illuminate the sample with each of a plurality of wavelengths, a fluorescence detector positioned to receive and substantially simultaneously detect multiple wavelengths of light emitted by the sample for each of the plurality of excitation wavelengths, an absorption detector positioned to receive and detect light passing through the sample, and a computer in communication with the excitation light source, the fluorescence detector, and the absorption detector, the computer controlling the excitation light source to sequentially illuminate the sample with each of the plurality of wavelengths while measuring absorption and fluorescence of the sample based on signals received from the fluorescence and absorption detectors.

Abstract: A method and system for luminescence molecular imaging or tomography of a region of interest in a scattering medium is disclosed. The system comprises a non-linear luminescent marker material arranged in the scattering medium, one or more light sources positioned by at least one light source position for exciting said luminescent marker by excitation light emitted by said one or more light sources into an excitation volume, a detector at a luminescent light detection position detecting luminescence from said luminescent marker due to said excitation light, wherein said excitation light comprises pulsed excitation light.

Abstract: A system and method for authenticating an item, including a photoluminescent material disposed on or in a substrate and capable of absorbing an incident radiation from a radiation source and emitting an emitted radiation having a spectral signature with a decay time after removal of the radiation source, and a photoauthentication device capable of being disposed in contact with the substrate and including the radiation source and a camera, where, in connection with providing the incident radiation and measuring the emitted radiation, the photoauthentication device is translated across the substrate while the photoauthentication device is disposed in contact with the substrate, and after translation across or over the substrate and the radiation source is not providing the incident radiation, the photoauthentication device is static with respect to the substrate and the camera is disposed over the photoluminescent material emitting the emitted radiation when the emitted radiation is measured.

Abstract: High-accuracy magnetic measurement is performed by efficiently using nitrogen-vacancy pairs in all orientations. A magnetic measurement apparatus includes a diamond crystal and an image sensor. The diamond crystal has nitrogen-vacancy pairs. The image sensor detects the intensities of fluorescence generated by an exciting light applied to the diamond crystal by using a plurality of pixels. The nitrogen-vacancy pairs of the diamond crystal are made to one-to-one correspond to the pixels. The fluorescence generated by one nitrogen-vacancy pair is received by one pixel made to correspond to the nitrogen-vacancy pair.

Abstract: An instrument and a method for measuring the characteristics of particles in a sample. The instrument comprises a light source operable to provide a light beam and defining an illumination axis; a sample cell placed on the illumination axis; a scattered light detector positioned to receive scattered light along a detection path from a sample in the sample cell, the scattered light produced by the interaction of the light beam with the sample; and a filter changer positioned between the sample cell and the scattered light detector. The filter changer comprises at least one optical filter and an actuator. The actuator is operable to move each of the at least one optical filter between a first position in which the detection path does not pass through the optical filter, and a second position in which the detection path passes through the optical filter.

Abstract: The present application relates to devices and methods for generating light sheets and thin light beams with high Rayleigh lengths by using at least two interconnected meso-optical elements.

Abstract: Improved methods, systems and apparatus relating to wide field fluorescence and reflectance imaging are provided, including improved methods, systems and apparatus relating to removal of background signals such as autofluorescence and/or fluorophore emission cross-talk; distance compensation of fluorescent signals; and co-registration of multiple signals emitted from three dimensional tissues.

Abstract: In one aspect, the present teachings provide a system for performing cytometry that can be operated in three operational modes. In one operational mode, a fluorescence image of a sample is obtained by exciting one or more fluorophore(s) present in the sample by an excitation beam formed as a superposition of a top-hat-shaped beam with a plurality of beams that are radiofrequency shifted relative to one another. In another operational mode, a sample can be illuminated successively over a time interval by a laser beam at a plurality of excitation frequencies in a scanning fashion. The fluorescence emission from the sample can be detected and analyzed, e.g., to generate a fluorescence image of the sample. In yet another operational mode, the system can be operated to illuminate a plurality of locations of a sample concurrently by a single excitation frequency, which can be generated, e.g., by shifting the central frequency of a laser beam by a radiofrequency.

Abstract: Apparatuses, methods, and systems are disclosed to characterize the wear and measure temperature of a surface of a lining of a metallurgical container. Wear characterization is accomplished by a scanning device by making thickness measurements of the lining and surface temperature measurements are made using a two-color pyrometer integrated with the scanning device where surface temperature is measured by correlating a ratio of two light intensities to the surface temperature. A controller controls operation of the scanning device and the two-color pyrometer.

Abstract: Optical and chemical analytical systems and methods are provided herein. In one embodiment, a method includes exposing a mixture sample to electromagnetic radiation, the mixture sample including analytes, detecting responsiveness of one or more of the analytes to the electromagnetic radiation, calculating average responsiveness of the one or more of the analytes, and calculating a concentration of the one or more of the analytes in the mixture sample using the average responsiveness.

Abstract: The fluorescent light phantom device 1 is provided with: a phantom support 10 having fluorescent light phantom containers 1b, 1c, 1d and 1e; and fluorescent light phantoms 12, 13, 14 and 15 which are constituted of a medium that reproduces at least one of light scattering and light absorption of an object to be measured and a fluorescent coloring matter contained in the medium of a predetermined concentration and are stored in the fluorescent light phantom containers 1b, 1c, 1d and 1e.

Abstract: A power saving movement includes a center wheel and pinion driving a minute hand and minute detection wheel. A gear ratio of the center wheel/pinion with respect to the minute detection wheel is 1/M. The minute detection wheel has N minute detection portions which are disposed on the same rotation trajectory as the center wheel & pinion. N and M are integers. The minute detection wheel transmittable portions are disposed at an interval of 360°/N. A pair of the center wheel transmittable portions are disposed in parallel at an unequal angular interval of a center axle of the center wheel & pinion. An angular interval of the center wheel transmittable portions adjacent to each other in the circumferential direction of the center axle of the center wheel & pinion is set to magnification of 360°/(M×N).

Abstract: A three-dimensional high-resolution localization microscopy method including illuminating a sample by excitation radiation to excite fluorescence markers in the sample to luminesce, and imaging the sample in an image frame via imaging optics along an imaging direction, wherein the image frame contains images of the luminescing fluorescence markers, and the imaging optics have a plane of focus and an optical resolution. The excitation step and imaging steps are repeated multiple times to generate a plurality of image frames, wherein the excitation steps are performed to isolate the images of the luminescing fluorescence markers in each image frame for at least some of the luminescing fluorescence markers. The location of the corresponding fluorescence marker is determined in each instance in the generated plurality of image frames from the isolated images of the luminescing fluorescence markers, and a highly resolved total image is generated from the locations determined in this way.

Abstract: A method for automated microscopic analysis wherein the test protocol is obtained from interrogatable data affixed to each microscope slide. The method further comprises the algorithms that implement the test protocol.

Abstract: A device includes a diamond with one or more nitrogen vacancies, a light emitting diode configured to emit light that travels through the diamond, and a photo sensor configured to sense the light. The device also includes a processor operatively coupled to the photo sensor. The processor is configured to determine, based on the light sensed by the photo sensor, a magnetic field applied to the diamond.

Abstract: The present invention relates generally to a system and a method for thermo-optical measurements in a droplet of aqueous solution comprising particles of interest, the method comprising the following steps: providing the droplet of aqueous solution with a volume of less than 200 nl, wherein the aqueous solution is a first liquid and at least a part of the particles of interest are fluorescent particles; embedding the droplet of aqueous solution at least partly in a second liquid; irradiating a laser light beam into the droplet to obtain a spatial temperature distribution in the droplet around the irradiated laser light beam; exciting fluorescently said fluorescent particles and detecting fluorescence at least at one position or at around one position in the droplet or detecting the fluorescence distribution of said fluorescently excited particles, wherein said detection of fluorescence is performed at least once at a predetermined time after the start of the laser irradiation; and determining a characteristic

Abstract: A fluorescence detecting apparatus includes an excitation light applying section that applies excitation light to a protective film containing an absorbing agent. A photomultiplier tube detects fluorescence emitted from the absorbing agent due to absorption of the excitation light. A fluorescence passing filter removes light having wavelengths other than the wavelength of the fluorescence emitted from the absorbing agent, and a reflecting mirror having a reflecting surface reflects the fluorescence emitted from the protective film toward the photomultiplier tube. This reflecting surface is formed by a part of a curved surface forming a spheroid having first and second foci. The first focus is positioned at a target area of the protective film where the excitation light is applied, and the second focus is positioned at a light detecting element included in the photomultiplier tube.

Abstract: A microscope apparatus includes an objective and a photodetector, and projects a pupil of the objective onto a light-receiving surface of the photodetector. The microscope apparatus further includes an optical element that is arranged between the objective and the photodetector. The optical element has a plurality of refractive surfaces which are arranged in a direction orthogonal to an optical axis of the optical element and to which light fluxes emitted from the pupil of the objective at different angles are incident.

Abstract: An environment sensitive coating system is disclosed that includes a pressure sensitive component comprising a first portion of an oxygen sensitive light emitting material dispersed in an oxygen permeable binder; and a pressure reference component comprising a second portion of the oxygen sensitive light emitting material dispersed in an oxygen impermeable binder.

Abstract: A method is disclosed for analyzing a thin tissue sample and adapted to be supported on a slide. The tissue sample may be placed on a slide and exposed to one or more different exogenous fluorophores excitable in a range of about 300 nm-200 nm, and having a useful emission band from about 350 nm-900 nm, and including one or more fluorescent dyes or fluorescently labeled molecular probes that accumulate in tissue or cellular components. The fluorophores may be excited with a first wavelength of UV light between about 200 nm-290 nm. An optical system collects emissions from the fluorophores at a second wavelength, different from the first wavelength, which are generated in response to the first wavelength of UV light, to produce an image for analysis.

Abstract: The present invention provides seeded rod (SR) nanostructure systems including an elongated structure embedded with a seed structure being a core/shell structure or a single-material rod element. The SR systems disclosed herein are suitable for use in a variety of electronic and optical devices.

Abstract: A flow cytometer including a laser, indexing structure, adjustment structure, and sensor structure. The cytometer is conventionally used with a removable microfluidic cassette, which is installed at a first position that is enforced by the indexing structure. The adjustment structure changes a relative position between an interrogation aperture of the cassette and the laser beam. Feedback from the sensor structure is used to optimize propagation of the laser through the interrogation aperture to reduce (and hopefully eliminate) autofluorescence caused by beam impingement onto the cassette.

Abstract: Embodiments of the present invention are directed to imaging technologies, and, in particular, to an imaging system that detects relatively weak signals, over time, and that uses the detected signals to determine the positions of signal emitters. Particular embodiments of the present invention are directed to methods and systems for imaging fluorophore-labeled samples in order to produce images of the sample at resolutions significantly greater than the diffraction-limited resolution associated with optical microscopy. Embodiments of the present invention employ overlapping-emitter-image disambiguation to allow data to be collected from densely arranged emitters, which significantly decreases the data-collection time for producing intermediate images as well as the number of intermediate images needed to computationally construct high-resolution final images. Additional embodiments of the present invention employ hierarchical image-processing techniques to further resolve and interpret disambiguated images.

Abstract: System and method for optical tomographic imaging optimizing analysis of time-domain data as a result of combination of lifetime multiplexing of low-cross-talk asymptotic photons with highly spatially resolved early photons. The tomographic data reconstruction employs a decay amplitude-based asymptotic approach and a matrix equation with a weight matrix that includes two different portions respectively representing time domain sensitivity functions and continuous-tomography weight matrices. System may employ a lifetime fluorescent tomography imaging system.

Abstract: A device for converting wavelength and a projection light source are disclosed in the present disclosure. According to an example, the device may comprise a roller, a drive motor, a first reflector, a support shaft, a drive module and a dichroic film. An axis of the support shaft may be coincided with an axis of the roller. The support shaft may be not rotated with rotation of the roller. The drive motor may drive the roller rotate around the support shaft and the drive module may drive the roller to move back and forth along an axial direction of the support shaft. So excitation light reflected by the first reflector may be incident uniformly on the sidewall of the roller. Therefore, fluorescent powder conversion efficiency may be improved compared with that of the excitation light incident on a fixed area.

Abstract: A fluorescence and phosphorescence detection device includes a fluorescence and phosphorescence sensor, a data acquiring unit, and an emission detection unit. The fluorescence and phosphorescence sensor includes a light source that emits an excitation light of a predetermined wavelength, and a photodetection unit that detects fluorescence emission and phosphorescence emission excited from the paper sheet by the excitation light. The data acquiring unit acquires a time-series waveform of a signal outputted from the fluorescence and phosphorescence sensor in response to the detection of the emission in the photodetection unit. The emission detection unit detects the fluorescence emission from the time-series waveform of a period in which the excitation light is emitted from the light source and detects the phosphorescence emission from an attenuation curve appearing on the time-series waveform of a period in which emission of the excitation light from the light source is stopped.

Abstract: The present invention relates to a method for detecting time-resolved fluorescence based on a principle of phase balanced frequency multiplication modulation. A stimulating light source modulated by using a baseband signal acts on a to-be-measured target to trigger fluorescence, so that the fluorescence intensifies and decays periodically; then, a frequency-doubled square signal is used to control a sampling period and divide an ascending period of the fluorescence into two and a decay period of the fluorescence into two; after independent sampling is performed separately, sampling differences of the two parts are separately calculated and then added to obtain an intensity representative value of a fluorescence signal and to obtain a concentration value of the to-be-measured target.

Abstract: A cold-test system for testing a camera at a cold temperature includes a test-chamber with a window, a nozzle, and a recirculator. The window is installed in the test-chamber so the camera is provided with a view of a test-target outside of the test-chamber. The nozzle is located proximate to a first edge of the window. The nozzle is configured to direct air from an air-source in a first direction and adjacent to an outside-surface of the window. The recirculator is located proximate to a second edge of the window opposite the first edge. The recirculator is configured to receive the air moving in the first direction and re-direct that air in a second direction opposite the first direction and adjacent to the air moving in the first direction opposite the window. The nozzle and the recirculator cooperate to form a bi-directional air-curtain proximate to the outside-surface.

Abstract: To detect a fluorescent light and a phosphorescent light emitted from a paper sheet with a high precision, a fluorescence and phosphorescence detecting apparatus includes two sensor units. Each sensor unit includes a light source that emits an ultraviolet light on the paper sheet, an image sensor that captures images of the fluorescent light and the phosphorescent light excited on the paper sheet by irradiation of the ultraviolet light, and a light receiving lens that guides the fluorescent light and the phosphorescent light excited on the paper sheet to the image sensor. The two sensor units are arranged above and below the transport path and opposing each other across the transport path, and the light receiving lenses and the image sensors in the two sensor units are off-set along a transport direction of the paper sheet.

Abstract: A microscope device includes an illumination optical system that illuminates a specimen with a light sheet, and a stereo image capturing device that captures images of the specimen in a plurality of different directions in which a resolution based on a triangulation method in a Z direction orthogonal to a direction of one or a plurality of light sheets formed on the specimen by the illumination optical system is less than a thickness in the Z direction of light sheet illumination that comprises the one or the plurality of light sheets.

Abstract: An oxygen sensing system comprises a substrate structured to communicate optical signals. An oxygen sensing layer is disposed on the substrate and comprises an oxygen sensing molecule in a matrix in a first unexcited state and formulated to: (a) be excited by a first optical signal to move to a second state; (b) be quenched in the second state by oxygen; and (c) emit a second optical signal corresponding to an amount of oxygen. A protective layer, disposed on the oxygen sensing layer, includes at least one of i) an oleophobic layer and ii) an anti-fouling layer. A controller is optically coupled to the substrate and structured to generate the first optical signal, receive the second optical signal and determine oxygen concentration from the second optical signal.

Abstract: An imaging flow cytometry apparatus and method which allows registering multiple locations across a cell, and/or across multiple flow channels, in parallel using radio-frequency-tagged emission (FIRE) coupled with a parallel optical detection scheme toward increasing analysis throughput. An optical source is modulated by multiple RF frequencies to produce an optical interrogation beam having a spatially distributed beat frequency. This beam is directed to one or more focused streams of cells whose responsive fluorescence, in different frequencies, is registered in parallel by an optical detector.

Abstract: Disclosed are a light-emitting diode and a method for manufacturing a light-emitting diode. The method includes: a base layer; a circuit layer formed on the base layer; a light-emitting chip formed on the circuit layer; electrode pads formed on the base layer and electrically connected to the light-emitting chip so that the electrode pads and the circuit layer and the light-emitting chip are spaced from each other by first spacing distances and the electrode pads and the circuit layer and the light-emitting chip define therebetween first grooves, where an altitude of the electrode pad is equal to an altitude of the light-emitting chip; and a phosphor powder contained package layer formed on the light-emitting chip and the electrode pads and filled into the first grooves between the electrode pads and the circuit layer to form a uniform dome shape.

Abstract: Method of optical microscopy by scanning a sample containing an excitable species, the method comprising: directing a first and a second light beam onto respective, partially overlapped areas of the sample, wherein the first light beam is provided for exciting members of the excitable species, and the second light beam is provided for reducing the number of excited members; detecting an optical signal coming from the sample, comprising a main component and a spurious component, during consecutive first and second time gates, the first time gate being provided for detecting the optical signal for a time interval during which the main component and the spurious component are both present, and the second time gate being provided for detecting the optical signal for a time interval during which the main component tends to or is zero; processing the detected optical signal to separate its main component.

Abstract: Disclosed are embodiments that relate to the deployment of a glucose sensor comprising an optical fiber into a physiological fluid, wherein the optical fiber has disposed along a distal region thereof a chemical indicator system comprising a fluorophore and a glucose binding moiety immobilized within a hydrogel, wherein the components of the chemical indicator system are in a dry state before deployment. Also disclosed is a one-point in vivo calibration of the chemical indicator system based on an independently measured glucose concentration.

Abstract: Embodiments of the present invention generally relate to methods for inspecting wafers. After a brick is sliced into a plurality of bare wafers, a two-dimensional (2D) photoluminescence (PL) image of each wafer is taken, the PL images of the wafers in sequential order (i.e., the sequence of the wafers as they are sliced from the brick) are then combined to construct a three-dimensional (3D) model of the brick that highlights similar regions in the brick.

Abstract: To minimize cross talk in systems and methods for detecting two or more different optical signals emitted from each of a plurality of reaction receptacles, an excitation signal associated with each of the optical signals has a known excitation frequency, and any detected signal having a frequency that is inconsistent with the excitation frequency is discarded. The receptacles are moved relative to optical sensors configured to detect each unique optical signal from an associated receptacle, and to further minimize cross talk, the optical sensors are arranged so that only one reaction receptacle at a time is in a signal detecting position with respect to one of its associated optical sensors, and the optical sensors are grouped by the optical signal they are configured to detect so that a first optical signal is detected from each of the reaction receptacles before a second optical signal is detected from the reaction receptacles.

Abstract: The present invention relates to the use of nanosystems as non deactivable security markers comprising metal atomic quantum clusters (AQCs) of at least two different size distributions encapsulated in a cavity with an inner diameter less than or equal to approximately 10 nm. These nanosystems are luminescence, particularly fluorescence after external excitation. The invention also relates to security documents, articles or elements incorporating these markers as well as to a method and a system for detecting the same.

Abstract: Disclosed herein is a protective film detecting method of detecting the formed condition of a protective film formed on the front side of a workpiece. The protective film detecting method includes a fluorescence intensity measuring step of forming a plurality of reference protective films having different thicknesses on the front sides of a plurality of reference workpieces, next applying excitation light absorbable by an absorbing agent contained in each reference protective film to each reference protective film, and next measuring the intensity of fluorescence emitted from the absorbing agent due to the absorption of the excitation light, and a threshold deciding step of deciding a threshold of the fluorescence intensity corresponding to a desired one of the different thicknesses of the reference protective films according to the fluorescence intensity measured above.

Abstract: Provided are compositions comprising modified DNA polymerases that exhibit improved photostability compared to the parental polymerases from which they were derived. Provided are methods for generating enzymes, such as DNA polymerases, with the aforementioned phenotype. Provided are methods of using polymerases with increased resistance to photodamage to make a DNA or to sequence a DNA template.

Abstract: Provided is an optical microscope capable of maintaining an objective lens in a focus state without adversely affecting a captured image even when a substance with a weak emission is used as an observation target object. The optical microscope includes an observation optical system capable of capturing, with an image pickup device (10), an optical image of an observation target object (1) that has been obtained with an objective lens (5), and sending out the optical image as an image signal; and an autofocus device that adjusts a focal position of the objective lens based on autofocus light emitted from a focusing light source (11) so as to focus on the observation target object, wherein the autofocus light is emitted from the focusing light source during a non-capturing period during which the image pickup device is not capturing an image of the observation target object.

Abstract: A spectroscopy system includes detectors configured to obtain detection spectrums of respective detection areas that are located at different positions of an object; and an information processor configured to obtain a target spectrum of a target area by using position information of the detection areas and the detection spectrums obtained by the detectors.

Abstract: A novel method for enhancing (for improved average power), stabilizing (to reduce rms noise), and wavelength tuning supercontinuum generation is described for numerous applications including but not limited to metrology, spectroscopy, and biomedical imaging. What is described is a way of enhancing only a small bandwidth portion of the optical spectrum by means of self-seeding in the supercontinuum. This cost-effective method allows for wavelength tuning while maintaining the enhancement.

Abstract: Provided is a method for detecting a target particle that is a method for detecting a non-luminescent target particle dispersed and randomly moving in a sample solution using an optical system of a confocal microscope or multi-photon microscope, having: (a) preparing a sample solution containing target particles, and labeling particles of which the average outer diameter is less than 15% of the diameter of a photodetection region of the optical system, binding two or more molecules of the labeling particles per molecule of the target particles in the sample solution, and forming a non-luminescent complex of which the outer diameter is 15% or more of the diameter of the photodetection region; and, (b) calculating the number of molecules of the complex in the sample solution prepared in the (a) using an inverse scanning molecule counting method.

Abstract: Provided is a method for detecting a target particle that is a method for detecting a non-luminescent target particle dispersed and randomly moving in a sample solution using an optical system of a confocal microscope or multi-photon microscope, having: (a) preparing a sample solution containing target particles, and labeling particles of which the average outer diameter is less than 15% of the diameter of a photodetection region of the optical system, binding two or more molecules of the labeling particles per molecule of the target particles in the sample solution, and forming a non-luminescent complex of which the outer diameter is 15% or more of the diameter of the photodetection region; and, (b) calculating the number of molecules of the complex in the sample solution prepared in the (a) using an inverse scanning molecule counting method.

Abstract: A nanoparticle-based system for enhancement of emitted light inside a medium. The nanoparticle-based system includes a phosphorescent material which, upon activation, produces the emitted light, and a metallic structure attached to the phosphorescent material. The metallic structure has a surface plasmon resonance which resonates at a frequency which spectrally overlaps with one or more frequencies of the emitted light.

Abstract: A flow cytometer including a laser, indexing structure, adjustment structure, and sensor structure. The cytometer is conventionally used with a removable microfluidic cassette, which is installed at a first position that is enforced by the indexing structure. The adjustment structure changes a relative position between an interrogation aperture of the cassette and the laser beam. Feedback from the sensor structure is used to optimize propagation of the laser through the interrogation aperture to reduce (and hopefully eliminate) autofluorescence caused by beam impingement onto the cassette.

Abstract: A fluorescence-spectrum-recording unit, a fluorescence-image-acquiring unit, and a fluorescent-dye-density-calculating unit are comprised, the calculating unit calculates the densities D1 to Dm of fluorescent dyes 1 to m in each of all the pixels and in all of the pixels with the following equation, and, when there exists a pixel in which one of the calculation values of these densities is smaller than 0, a set value larger than the calculation value smaller than 0 is substituted for the density the calculation value of which is smaller than 0 in the equation, and the densities of the other fluorescent dyes are recalculated, relative to the pixel: ( D ? ? 1 ? Dm ) = ( a ? ? 1 ? ( ? ? ? 1 ) … am ? ( ? ? ? 1 ) ? ? ? a ? ? 1 ? ( ? ? ? n ) … am ? ( ? ? ? n ) ) - 1 ? ( I all ? ( ? ? ? 1 ) ? I all ? ( ? ? ? n ) ) where a1 (?1) to am (?n) denote the coefficients for the flu

Abstract: A high-resolution fluorescence image in which an afterimage is suppressed is obtained, even when a fluorescence detection interval is shortened. Provided is a scanning observation apparatus including a scanning unit that spatially scans pulsed excitation light emitted from a light source at prescribed time intervals on a specimen; a fluorescence detecting unit that detects fluorescence generated by exciting a fluorescent substance inside the specimen with the excitation light scanned by the scanning unit, in synchronization with the emission of the excitation light; and a fluorescence correcting unit that subtracts, from a fluorescence intensity detected by the fluorescence detecting unit, an afterimage fluorescence component calculated on the basis of time-sequential fluorescence detected by the fluorescence detecting unit prior thereto, at each scanning position, to correct the fluorescence intensity at the scanning position.