I.s.s. (usa), Inc.

University of California

University of Illinois

Research Corporation Technologies, Inc.

Ykk Corporation

Enrico Gratton

Beniamino Barbieri

Maria Angela Franceschini

Sergio Fantini

Alexander Dvornikov

William Mantulin

Michelle A Digman

Guido Motolese

Abdel Tahari

Brett A Feddersen

Luisa Marsili

Martin J vandeVen

David W Piston

Kaveh Azartash

William W Mantulin

Francesco Cutrale

Martin VandeVen

John Maier

Chiara Stringari

Luca Lanzano

Scott A Walker

Peter So

Chen-Yuan Dong

Peter Donovan

Viera Crosignani

Enrico D'Amico

Enrico D'AMICO

An imaging system comprises an excitation light source, a directing element positioned to direct light from the excitation light source toward a sample, a detector configured to measure incoming light from the sample, a filter cavity positioned between the sample and the detector, a first filter configured to be inserted into the filter cavity, a sine filter configured to be inserted into the filter cavity, and a processing unit communicatively connected to the detector, configured to receive image data from the detector to form an image. Methods of constructing a hyperspectral image of a sample are also described.

System and method for hyperspectral imaging in highly scattering media by the spectral phasor approach using two filters

A label-free imaging method to monitor stem cell metabolism discriminates different states of stem cell as they differentiate in a living tissues. We use intrinsic fluorescence biomarkers and the phasor approach to Fluorescence Lifetime Imaging Microscopy (FLIM). We identify and map intrinsic fluorophores such as collagen, retinol, retinoic acid, flavins, nicotinamide adenine dinucleotide (NADH) and porphyrin. We measure the phasor values of germ cells ingerm line. Their metabolic fingerprint cluster according to their differentiation state, reflecting changes in FAD concentration and NADH binding during the differentiation pathway. The phasor approach to lifetime imaging provides a label-free, fit-free and sensitive method to identify different metabolic state of cells during differentiation, to sense small changes in the redox state of cells and may identify symmetric and asymmetric divisions and predict cell fate.

Phasor method to fluorescence lifetime microscopy to discriminate metabolic state of cells in living tissue

An apparatus for inclined single plane Illumination microscopy of a sample includes a laser for launching excitation light beams at a plurality of wavelengths, a laser beam expander, an injection arm optically coupled to the laser beam expander, a conventional back-to-back microscope system, a universal dichroic mirror optically coupled to the injection arm to direct the excitation light beams into the conventional back-to-back microscope onto a sample plane in an imaging plane, and to receive fluorescence light from the sample, a universal optical adaptor optically coupled to the universal dichroic mirror, a re-imaging component optically coupled to the universal optical adaptor; and a camera output connector optically coupled to the re-imaging component, where the laser beam expander, injection arm, universal optical adapter, re-imaging component, and camera are combined in a modular unit which is arranged and configured to be coupled to the conventional back-to-back microscope.

Apparatus and method for an inclined single plane imaging microscope box (iSPIM box)

An optical imaging method based on a feedback principle in which the specific scan pattern is adapted according to the shape of the sample. The feedback approach produces nanometer-resolved three dimensional images of very small and moving features in live cells and in a matter of seconds. Images of microvilli in live cultured opossum kidney cells expressing NaPi co-transporter proteins with different GFP constructs and images of cell protrusions in a collagen matrix are produced with a resolution of about 20 nm. Along cell protrusions in three dimensional cellular adhesions could be identified to the extracellular matrix.

Nanometer-scale optical imaging by the modulation tracking (MT) method

An apparatus and method for in-depth fluorescence imaging using two-photon fluorescence imaging in turbid media. The apparatus includes a detector which can significantly enhance the use of a detection method that allows to efficiently collect scattered fluorescence photons from a wide area of the turbid sample. By using this detector it is possible to perform imaging of turbid samples, simulating brain tissue at depths up to 3 mm, where the two-photon induced fluorescence signal is too weak to be detected by previous means used in conventional two-photon microscopy. The detector separates the excitation and detection optics which allows a more efficient collection of fluorescence and enhancing the possible imaging depth.

Apparatus and method for light emission detection for in-depth imaging of turbid media

A system and method is provided for improved fluorescence decay time measurement. A digital heterodyning technique is disclosed in which a photon detector is sampled at a rate slightly faster than a digitally pulsed excitation signal. A resulting cross correlation frequency is low enough to be read by inexpensive electronics such as by a field programmable gate array. Phase information in the signal provides correlation with corresponding photon detections.

System and method for digital parallel frequency fluorometry

Tear film stability has an important role in the quality of vision. A system and method for performing Fluctuation Analysis of Spatial Image Correlation (FASIC) provides for a non-invasive system and method for evaluating the dynamics of the tear film surface using spatial autocorrelation analysis. With FASIC, a series of images are obtained using illumination and a camera. The spatial autocorrelation is calculated for image frames produced by the camera. A sinusoidal background appears in this correlation together with other features. The changes in the sinusoidal background of the spatial autocorrelation is extracted and monitored over time. The spatial period of this sinusoidal background correlates with the thickness of the tear film. In this regard, one is able to derive the tear film thickness from the period of this sinusoidal background.

Method and device for tear film analysis

The invention provides methods and devices for detecting, identifying, classifying and characterizing particles in a fluid sample. Optical analyzers are provided having a rotating and/or translating sample container for measuring the concentrations of fluorescent particles present in very low concentrations and for characterizing fluorescent particles on the basis of size, shape, diffusion constant and/or composition. Scanning optical analyzers are provided using pattern recognitions data analysis techniques and multichannel detection.

Methods and devices for characterizing particles in clear and turbid media

A fluorescence spectrometer comprises a laser and at least one beam splitter positioned to receive a light beam from the laser and to divide it into several first light beam portions. Dichroic mirrors are positioned to separately receive the first light beam portions and to reflect the beam portions at an angle to the first light beam portions. Transparent chambers are provided for holding the samples. Objective lens systems are respectively positioned in the path of the reflected beam portions to respectively focus each reflected beam portion to a point within one of the separate transparent chambers. Lenses are positioned to receive fluorescence from a sample for testing within the transparent chambers and to respectively focus the fluorescence at pin holes in opaque partitions. The lenses are positioned to receive the fluorescence, which passes back through the objective lens system and the dichroic mirror. Light detectors are each respectively positioned adjacent to one of the partitions, with one of the partitions respectively positioned between each of the lens and the light detectors to permit each light detector to sense fluorescence through the pin hole. Electronics are provided to receive and process signals from each light detector. Structure is provided to permit high speed data collection from a large number of samples in separate, transparent chambers.

Rapid high throughput spectrometer and method

The present invention involves a time-resolved measurement method for the real time, non-invasive, simultaneous measurement of time-varying and other hemoglobin compartment saturation. This capability achieves absolute pulse oximetry and oximetry for tissue, without calibration based on a population of healthy people. Calculations conducted by the invention use quantitative measurement of tissue absorption spectrum for tissue saturation, and an amplitude of absorption oscillations for the time-varying hemoglobin compartments at various wavelengths. The invention illuminates tissue and senses light at predetermined distances apart on the tissue to be measured. Intensity and phase data are acquired from source-detector pairs to calculate absolute tissue optical properties from time-resolved measurement data, namely, a reduced scattering coefficient and an absorption coefficient. To determine time-varying hemoglobin compartment saturation, an amplitude is quantitatively calculated of absorption oscillations correlating variations of an average intensity of the source and detector pair by using the time-resolved measurement data.

Method for measuring absolute saturation of time-varying and other hemoglobin compartments

The quantitative determination of various materials in highly scattering media such as living tissue may be determined in an external, photometric manner by the use of a plurality of light sources positioned at differing distances from a sensor. The light from said sources is amplitude modulated, and, in accordance with conventional frequency domain fluorometry or phosphorimetry techniques, the gain of the sensor is modulated at a frequency different from the frequency of the light modulation. Data may be acquired from each of the light sources at differing distances at a frequency which is the difference between the two frequencies described above. From these sets of data from each individual light source, curves may be constructed, and the slopes used to quantitatively determine the amount of certain materials present, for example glucose, oxyhemoglobin and deoxyhemoglobin in living tissue.

Photosensor with multiple light sources

A scanning fluorescence lifetime microscope measures modulation and phase in fluorescence emission stimulated by spatially overlapped pump and probe beams operating at different frequencies. A pump laser modulated at a first frequency is focused onto a diffraction limited spot to excite a fluorescent sample under study. Simultaneously, a probe laser modulated at a second frequency is focused onto the same spot to induce a stimulated fluorescence emission in response to the optically combined output of the pump and probe laser light. Fluorescence emitted from the sample produces a cross-correlation signal which is dependent upon the spatial overlapping of the pump and probe beams at the focal point thereby producing a beneficial axial sectioning effect. Choosing a small differency frequency between the modulation of the first and second laser sources produces a low frequency cross correlation signal even when the modulation frequencies of the pump and probe lasers are very high. A signal processor obtains modulation and phase information from the low frequency cross correlation fluorescence signal allowing monitoring of even ultrafast fluorescence phenomena induced by high frequency modulation of the pump and probe lasers.

Growing community of inventors

San Clemente, CA, United States of America

Enrico Gratton