Multiphoton Microscopy Wavelength Tuning for Deep Tissue Imaging
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Solution Overview
Problem
Multiphoton microscopy is limited by light absorption, scattering, and the power of the incident beam, restricting imaging depth and photochrome excitation possibilities due to the choice of wavelengths used.
Innovation Solution
A device comprising a laser source emitting a central wavelength between 1010 nm and 1050 nm, a spectral supercontinuum generator, an optical parametric amplification system, and a second harmonic generator, producing synchronous beams with central wavelengths between 1030 nm, 1300 nm, and 1700 nm, which are within windows of lesser water absorption and scattering, allowing for enhanced imaging depth and photochrome excitation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of energy
If a femtosecond laser emitting ultrashort pulses with a wavelength between 700 nm and 1100 nm is used, then the absorption of water and other constituents is minimized, but the imaging depth remains limited by light scattering in the tissues
Solution Approach 1:
The invention changes the wavelength parameter from the conventional 700-1100 nm range to 1300 nm or 1700 nm, which are located in water absorption windows. This parameter change simultaneously reduces both water absorption and light scattering, thereby increasing imaging depth while maintaining low energy loss
2Length of moving object
If a femtosecond laser emitting ultrashort pulses with a wavelength centered on 1300 nm or 1700 nm is used, then the imaging depth is increased, but the choice of accessible photochromes is limited
Solution Approach 1:
The invention segments the excitation process by using multiple discrete wavelengths (1300 nm and 1700 nm) instead of a single wavelength. Each wavelength can excite different photochromes, thereby expanding the range of accessible photochromes while maintaining deep imaging capability
Solution Approach 2:
The invention makes the microscopy system multi-functional by enabling excitation of multiple types of photochromes using two different wavelengths. This allows the same system to access a broader range of fluorescent probes without requiring separate specialized systems
3Adaptability or versatility
If multiple wavelengths are used to access different photochromes, then the imaging possibilities are expanded, but the system complexity increases
Solution Approach 1:
The invention merges multiple wavelength sources into a single femtosecond laser system operating at 1300 nm or 1700 nm. By using the nonlinear optical properties of tissue and appropriate fluorophores, the system achieves multi-wavelength excitation capability through a single laser source, thereby expanding imaging possibilities while minimizing system complexity
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The device enables deeper tissue imaging and broader photochrome excitation possibilities by generating beams with wavelengths that minimize absorption and scattering, offering multiple excitation options and improving imaging capabilities in multiphoton microscopy.
Implementation Method 1
a spectral supercontinuum generator downstream of the laser source, generating a second beam with a central wavelength lying between 1670 nm and 1730 nm from a part of the first beam
Implementation Method 2
an optical parametric amplification system downstream of the spectral supercontinuum generator, generating a third beam with a central wavelength lying between 2545 nm and 2690 nm from at least a part of the second beam and a part of the first beam
Implementation Method 3
a second harmonic generator downstream of the optical parametric amplification system, the second harmonic generator generating a fourth beam with a central wavelength lying between 1270 nm and 1345 nm from at least a part of the third beam
Implementation Method 4
Multiphoton microscopy makes it possible to image biological tissues to a certain depth non-invasively with a micrometric three-dimensional resolution. It consists in exciting fluorochromes at a given focal point with several photons simultaneously
Data Source
AI summary
The invention relates to a device comprising:a laser source emitting a first beam with a central wavelength λ1 lying between 1010 nm and 1050 nm,a spectral supercontinuum generator downstream of the laser source, generating a second beam with a central wavelength λ2 lying between 1670 nm and 1730 nm from a part of the first beam,an optical parametric amplification system downstream of the spectral supercontinuum generator, generating a third beam with a central wavelength λ3 lying between 2540 nm and 2690 nm from at least a part of the second beam and a part of the first beam, anda second harmonic generator downstream of the optical parametric amplification system, the second harmonic generator generating a fourth beam with a central wavelength λ4 lying between 1270 nm and 1345 nm from at least a part of the third beam.