Microscope Excitation Waveform for Second-Harmonic Detection
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Solution Overview
Problem
Existing microscopes combining saturated excitation (SAX) and multiphoton excitation face limitations in frequency range detection, leading to increased time or cost for creating observation images due to the need to detect high-order harmonics.
Innovation Solution
A microscope device and image acquisition method using an excitation beam with a temporal waveform that is the n-th root of a linear function of a sine wave, allowing detection of lower-order harmonics like the second or third harmonic, reducing the need for frequency reduction or high-cost devices.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If an excitation beam with sinusoidal temporal waveform is used for n-photon excitation, then fluorescence intensity is generated, but high-order harmonics (third or fifth) must be detected which increases device cost or reduces imaging speed
Solution Approach 1:
The patent changes the temporal waveform parameter of the excitation beam from a standard sine wave to a square wave. This parameter change transforms the harmonic content of the fluorescence signal, allowing the second harmonic to be detected instead of requiring third or fifth harmonics. The square wave excitation creates a fluorescence temporal waveform that contains strong second harmonic components, which can be detected by devices with lower frequency range limits, thus reducing device complexity and cost.
2Measurement precision
If the frequency of the excitation beam is reduced to detect high-order harmonics, then detection becomes possible, but the time required to create an observation image is increased
Solution Approach 1:
By changing the temporal waveform from sine to square wave, the patent enables detection at the second harmonic frequency instead of requiring third or fifth harmonics. This allows the excitation beam frequency to be maintained at higher values, thereby maintaining faster image creation speed while still achieving the spatial resolution benefits of saturated excitation microscopy.
3Measurement precision
If a device with higher frequency range is introduced to detect high-order harmonics, then observation images can be created, but the cost is increased
Solution Approach 1:
The patent changes the excitation waveform to a square wave, which transforms the fluorescence signal to contain detectable second harmonic components. This allows the use of devices with lower frequency range specifications, reducing the need for expensive high-frequency detectors and thereby lowering the overall device cost while maintaining spatial resolution improvement.
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
Enables efficient creation of observation images using lower-order harmonics, avoiding time increases and cost escalation by utilizing a device with a lower frequency range.
Implementation Method 1
an object to be observed is irradiated with an excitation beam having a sinusoidal temporal waveform, the object to be observed is excited to output fluorescence
Implementation Method 2
an object to be observed is irradiated with long-wavelength ultrashort-pulse light such as near-infrared light as an excitation light beam to cause multiphoton excitation such as two-photon excitation to occur in the object to be observed
Implementation Method 3
The optical system irradiates the object to be observed with the excitation beam output from the excitation beam output unit
Implementation Method 4
The harmonic detector detects a second harmonic included in a temporal waveform of a light intensity of fluorescence generated in the object to be observed
Data Source
AI summary
A microscope device includes an excitation beam output unit, an optical system, and a harmonic detector. The excitation beam output unit outputs excitation beam. A temporal waveform of a light intensity of the excitation beam includes an n-th root (n is an integer of 2 or more) of a linear function of a sine wave. A maximum value of the light intensity of the excitation beam is higher than a saturation excitation intensity of an object to be observed. The optical system irradiates the object to be observed with the excitation beam output from the excitation beam output unit. Fluorescence is generated in the object to be observed due to an n-photon excitation by the irradiation with the excitation beam. The harmonic detector detects a second harmonic included in a temporal waveform of a light intensity of the fluorescence.