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Precision Imaging with Two-Photon Microscope Technology

Introduction

In the fields of biotechnology, medical research, and diagnostics, high-resolution imaging is crucial for understanding complex biological processes and discovering new therapies. However, traditional imaging techniques often fall short when it comes to deep tissue penetration and achieving molecular-level detail without damaging live samples. Our patented two-photon microscope with spectral resolution offers a cutting-edge solution, providing unprecedented imaging depth and clarity that can transform the way researchers visualize biological structures in live tissue.

Current Gaps in Imaging Technology

Conventional microscopes, while useful for many research purposes, struggle with limitations in penetration depth and resolution when applied to thick or live tissue samples. Additionally, imaging live tissues can result in sample damage due to high-intensity light exposure. This is particularly problematic in fields like neuroscience and cancer research, where precise, non-invasive imaging is essential for observing biological processes in real time. Furthermore, standard imaging systems lack spectral resolution, making it difficult to distinguish between different molecular signatures within a sample.

These limitations create barriers for researchers who require more detailed and precise imaging technologies to gain meaningful insights from their work. The need for a high-performance system capable of imaging deep into live tissues, while maintaining both clarity and spectral resolution, is clear across many scientific disciplines.

A Breakthrough in Spectral Imaging with Two-Photon Microscopy

Our patented two-photon microscope with spectral resolution addresses these challenges by enabling high-resolution, deep tissue imaging while minimizing sample damage. Two-photon excitation allows for deeper penetration into tissues with less photodamage, making it ideal for imaging live cells, tissues, and even entire organisms. The addition of spectral resolution further enhances the technology by allowing researchers to separate and analyze distinct molecular signals, providing a more detailed view of complex biological structures.

This innovative system can be used in a wide range of applications, from mapping neural networks in the brain to studying the behavior of cancer cells. Its non-invasive nature makes it a powerful tool for live imaging in real-time, offering researchers greater flexibility and accuracy in their studies. Additionally, the spectral capabilities enable the differentiation of various fluorophores or molecular markers, unlocking new possibilities in multi-modal imaging.

Key Advantages

  • Deep Tissue Penetration: Two-photon excitation allows for clear imaging of live tissues without damaging the sample.
  • Spectral Resolution: This technology can separate molecular signals, enabling the study of multiple markers in a single sample.
  • Non-Invasive Imaging: The system is ideal for live imaging, reducing photodamage and allowing for real-time observations.
  • Wide Range of Applications: From neuroscience to cancer research, this microscope opens up new possibilities for advanced research.

A Leap Forward in Imaging Precision

Licensing this two-photon microscope technology with spectral resolution provides research institutions, biotechnology companies, and pharmaceutical developers with a highly advanced imaging tool. With applications spanning from basic research to drug discovery, this technology enables scientists to visualize biological processes in unprecedented detail, offering a significant edge in the race to make new breakthroughs.

Please see terms and conditions
A microscope for generating an image of a sample, the microscope includes a light source for generating a pulsed light; a scanning mirror receiving the pulsed light and transmitting the pulsed light to the sample to be imaged causing the sample to emit energy; a dispersive element that receives the emitted energy from the sample, disperses the energy into its spectral elements and transmits the spectrally dispersed energy; and a camera that generates a spectrally resolved image of the sample based on the spectrally dispersed energy from the dispersive element. Also described is a method of generating spectrally resolved images of the sample.

1. A microscope for generating a multi-dimensional, spectrally resolved image of a dynamic sample, the microscope comprising:

a light source for generating a pulsed light;
a scanning mirror in one of a fully non-descanned arrangement and a half-descanned arrangement receiving the pulsed light and transmitting the pulsed light to the dynamic sample to be imaged, causing the sample to emit energy;
a dispersive element that receives the emitted energy from a single scan of the dynamic sample, disperses the energy into its spectral elements and transmits the spectrally dispersed energy; and
a camera that generates a multi-dimensional, spectrally resolved image of the dynamic sample based on the spectrally dispersed energy from the dispersive element using only the emitted energy from the single scan of the sample.
2. A microscope as set forth in claim 1 wherein the light source comprises a laser.
3. A microscope as set forth in claim 1 wherein the scanning mirror comprises a computer controlled x-y scanning mirror.
4. A microscope as set forth in claim 1 wherein the scanning mirror comprises a plurality of computer controlled mirrors, and wherein one of the mirrors is a short pass, dichroic mirror.
5. A microscope as set forth in claim 1 wherein the scanning mirror comprises a pair of computer controlled scanning mirrors and a long pass dichroic mirror positioned between the computer controlled scanning mirrors.
6. A microscope as set forth in claim 1 wherein the dispersive element comprises one of a transmission grating, and a prism.
7. A microscope as set forth in claim 1 wherein the camera comprises a charge coupled device camera.
8. A microscope as set forth in claim 1, and further comprising a round lens for focusing the light from the light source to a point.
9. A microscope as set forth in claim 1, and further comprising a cylindrical lens for focusing the light from the light source in a line.
10. A microscope as set forth in claim 1, and further comprising a broadband light source positioned beneath the sample to be imaged and a narrow band-pass filter positioned between the sample and the camera to provide transmission imaging of the sample.

11. A microscope for generating an image of a sample, the microscope comprising:

a laser light source for generating a pulsed light;
a computer controlled scanner receiving the pulsed light and scanning the pulsed light across the sample to be imaged, causing the sample to emit energy, wherein the scanner is part of one of a fully non-descanned arrangement and a half-descanned arrangement;
at least one of a transmission grating and a prism that receives the emitted energy from a single scan of the sample, disperses the energy into its spectral elements and transmits the spectrally dispersed energy; and
a charge coupled device camera that generates a spectrally resolved image of the sample based on the spectrally dispersed energy from the dispersive element using only the emitted energy from the single scan of the sample.

12. A method of generating an image of a sample having x and y dimensions and using a microscope having a laser light source, a computer controlled scanning mirror, a dispersive element and a camera, the method comprising:

generating laser energy;
scanning the laser energy across the sample to cause emission of light from the sample using a scanner in one of a fully non-descanned arrangement and a half descanning arrangement;
dispersing the light emitted from the sample into its spectral elements;
receiving the spectral elements of the dispersed light as a continuous spectrum; and
generating a spectrally resolved image of the sample at a given wavelength based on only a single scan of the sample.
13. A method as set forth in claim 12 wherein dispersing the light from the sample into its spectral elements comprises passing the light through the dispersive element.
14. A method as set forth in claim 13 wherein the dispersive element is one of a transmission grating, reflection grating and a prism.
15. A method as set forth in claim 12 wherein generating a spectrally resolved image of the sample at a given wavelength comprises acquiring a series of spectral images for each y-position on the sample and reconstructing an image from the components of each of the series of spectral images corresponding to the given wavelength.
16. A method as set forth in claim 12 and further comprising calibrating the microscope by generating fluorescence images of a sample that fluoresces uniformly across the sample and relating row numbers in the y direction with spectral elements of the emitted light from the sample.

17. A method of generating an image of a sample having x and y dimensions and using a microscope having a laser light source, a computer controlled scanning mirror, a dispersive element and a camera, the method comprising:

generating laser energy;
scanning the laser energy across the sample to cause emission of light from the sample using a scanner in one of a fully non-descanned arrangement and a half descanning arrangement;
dispersing the light emitted from the sample into its spectral elements by passing the light through one of a transmission grating and a prism; and
generating a multi-dimensional, spectrally resolved image of the sample at a given wavelength by acquiring a series of spectral images for each y-position on the sample and reconstructing an image from the components of each of the series of spectral images corresponding to the given wavelength using only the emitted energy from a single scan of the sample.
18. A microscope as set forth in claim 1 wherein the multi-dimensional, spectrally resolved image is a two-dimensional image of the dynamic sample, in which one dimension is a line scanned along a geometrical dimension of the dynamic sample and another dimension is a wavelength of the spectrally dispersed energy from the dispersive element.
19. A microscope as set forth in claim 1 wherein the dynamic sample is a biological sample.
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Title

Two-photon microscope with spectral resolution

Inventor(s)

Valerica Raicu, Russell Fung

Assignee(s)

UWM Research Foundation Inc

Patent #

7973927

Patent Date

July 5, 2011

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