Massachusetts Institute of Technology

National Taiwan University

National University of Singapore

Nanyang Technological University

Peter T C So

Timothy M Ragan

Karsten Bahlman

Ki-Hean Kim

Bevin Engelward

Zahid Yaqoob

Hayden Huang

Daekeun Kim

Lily Hsu Laiho

Dushan N Wadduwage

Chen-Yuan Dong

Hyung Min Kim

Hanry Yu

Ramachandra Rac Dasari

Christopher Rowlands

Jeon Woong Kang

Yongjin Sung

Vijay Raj Singh

Niyom Lue

Elly Nedivi

Jagath Rajapakse

Colin J R Sheppard

Dominika Lyzwa

Jong Kang Park

Yi Xue

Kalen Berry

Sunney I Chan

Yan Seng Elijah Yew

Vijay Singh

Hyuk-Sang Kwon

Heejin Choi

George Cragg

Systems and methods herein provide improved, high-throughput multiphoton imaging of thick samples with reduced emission scattering. The systems and methods use structured illumination to modify the excitation light. A reconstruction process can be applied to the resulting images to recover image information free of scattering. The disclosed systems and methods provide high throughput, high signal-to-noise ratio, and high resolution images that are depth selective.

Systems and methods to reduce scattering in temporal focusing multiphoton microscopy

Since the fat content of pork is a deciding factor in grading the quality of meat, the use of a noninvasive subcutaneous probe for real-time, in situ monitoring of the fat components is of importance to vendors and other interested parties. Fortunately, in situ, in vivo monitoring of subcutaneous fat can be accomplished with spatially offset Raman spectroscopy (SORS) using a fiber-optic probe. The probe acquires Raman spectra as a function of spatial offset. These spectra are used to determine the relative composition of fat-to-skin. The Raman intensity ratio varies disproportionately depending on the fat content, with variations in slope that are correlated to the thickness of the fat layer. Ordinary least square (OLS) regression using two components indicates that depth-resolved SORS spectra reflect the relative thickness of the fat layer.

Subcutaneous fat thickness measurement by Raman spectroscopy

Our high phase sensitivity wide-field phase cancellation interferometry system allows single-shot, label-free optical sensing of neural action potentials via imaging of optical path length changes. Single-shot sensing and monitoring of single neurons within a neural network should lead to a more comprehensive understanding neural network processing, which is beneficial for the advancement in the field of neuroscience as well as its biomedical applications and impact. Our system cancels the phase profile of the resting neuron from the phase profile of the spiking neuron, improving the sensitivity by two orders of magnitude. Using a detector with an extremely large well depth and an appropriately biased interferometer increases the sensitivity by another order of magnitude, yielding a measurement that is three orders of magnitude more sensitive than those possible with other microscopes.

Phase cancellation microscopy

Refractive index of biological specimens is a source of intrinsic contrast that can be explored without any concerns of photobleaching or harmful effects caused by extra contrast agents. This feature also contains rich information that can be related to the metabolism of cells at the cellular and subcellular levels. The present invention relates to systems and methods that can provide, without any moving parts, the 3-D refractive index map of continuously flowing biological samples in a micro-fluidic channel, for example.

3-D holographic imaging continuous flow cytometry

In the systems and methods of the present invention a multifocal multiphoton imaging system has a signal to noise ratio (SNR) that is reduced by over an order of magnitude at imaging depth equal to twice the mean free path scattering length of the specimen. An MMM system based on an area detector such as a multianode photomultiplier tube (MAPMT) that is optimized for high-speed tissue imaging. The specimen is raster-scanned with an array of excitation light beams. The emission photons from the array of excitation foci are collected simultaneously by a MAPMT and the signals from each anode are detected using high sensitivity, low noise single photon counting circuits. An image is formed by the temporal encoding of the integrated signal with a raster scanning pattern. A deconvolution procedure taking account of the spatial distribution and the raster temporal encoding of collected photons can be used to improve decay coefficient. We demonstrate MAPMT-based MMM can provide significantly better contrast than CCD-based existing systems.

Multifocal imaging systems and method

An imaging system is provided that includes a pulsed light source providing pulsed light and is applicable to both microscopes and endoscopes. One or more optical elements with certain dispersive properties are positioned to receive the pulsed light and apply selective dispersive properties to shift the focal plane according to the user and to produce two photon (2p) wide field uniform illumination and 2p wide field structured illumination for the purpose of improving the optical axial resolution and rejection of background signal. An imaging element receives the signal arising from the 2p wide field uniform illumination and 2p wide field structured illumination and produces a respective 3D resolved image of a sample.

High sensitivity temporal focusing widefield multiphoton endoscope capable of deep imaging

In accordance with preferred embodiments of the present invention, a method for imaging tissue, for example, includes the steps of mounting the tissue on a computer controlled stage of a microscope, determining volumetric imaging parameters, directing at least two photons into a region of interest, scanning the region of interest across a portion of the tissue, imaging a plurality of layers of the tissue in a plurality of volumes of the tissue in the region of interest, sectioning the portion of the tissue and imaging a second plurality of layers of the tissue in a second plurality of volumes of the tissue in the region of interest, detecting a fluorescence image of the tissue due to said excitation light; and processing three-dimensional data that is collected to create a three-dimensional image of the region of interest.

Systems and methods for volumetric tissue scanning microscopy

An imaging system is provided that includes a optical pulse generator for providing an optical pulse having a spectral bandwidth and includes monochromatic waves having different wavelengths. A dispersive element receives a second optical pulse associated with the optical pulse and disperses the second optical pulse at different angles on the surface of the dispersive element depending on wavelength. One or more focal elements receives the dispersed second optical pulse produced on the dispersive element. The one or more focal element recombine the dispersed second optical pulse at a focal plane on a specimen where the width of the optical pulse is restored at the focal plane.

3D two-photon lithographic microfabrication system

A multiphoton excitaion microscope for simultaneously detecting differently colored fluorescence materials on biochips includes a multiphoton excitation source, objectives, and a plurality of detection channels. The biochip is hybridized and labeled with fluorescence materials for expressing hybridized biological signals. The multiphoton excitation source is focused to a light spot on the biochip to excite the fluorescence materials bound thereon. After that, the fluorescence emission at different wavelengths from the different fluorescent materials can be detected by the plural detection channels.

Multiphoton excitation microscope for biochip fluorescence assay

The invention features a microscopy system for imaging a sample including: a substantially transparent optical block having an interface for positioning the sample adjacent the interface; a light source which during operation overlaps at least two optical beams at the interface, the source directing the beams into the block towards the interface at incident angles that cause the beams to reflect from the interface and establish an evanescent standing wave of electromagnetic energy that extends away from the interface and into the sample; and a detector which during operation records an image of the sample based on optical radiation emitted from the sample in response to the evanescent standing wave. The invention also features a method for determining a high-resolution image of a sample including: coupling an evanescent standing wave of electromagnetic energy into the sample; recording an image of the sample based on radiation emitted from the sample in response to the evanescent standing wave for each of multiple positions of the standing wave; and constructing the high-resolution image based on the multiple recorded images.

Standing wave total internal reflection imaging

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Boston, MA, United States of America

Peter T C So