Manipulated Excitation Beams for Wavefront-Corrected Microscopy
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Solution Overview
Problem
Current methods for correcting excitation radiation wavefront in laser scanning microscopy are slow and require high intensities, making them unsuitable for imaging radiation-sensitive and rapidly changing samples.
Innovation Solution
A method and device that focuses a first excitation radiation into a sample volume, detects and evaluates its wavefront using a detector, manipulates a second excitation radiation to correct deviations, and scans the sample with separated partial beams to acquire image data, allowing for local correction of wavefront errors and increased sensitivity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If adaptive optics methods are used to correct wavefront aberrations, then image quality is improved, but acquisition time increases and sample damage occurs due to repeated illumination
Solution Approach 1:
The patent performs wavefront measurement and correction in advance before the actual image acquisition. The method measures the wavefront aberration using a test beam, calculates the correction values, and applies the correction to the imaging beam before capturing the sample images. This preliminary correction eliminates the need for repeated measurements and adjustments during image acquisition, thereby reducing acquisition time and minimizing sample damage from repeated illumination.
Solution Approach 2:
The patent separates the wavefront measurement process from the image acquisition process. It uses a dedicated test beam path for wavefront measurement and a separate imaging beam path for sample observation. This segmentation allows the measurement and correction to be performed independently and in advance, without interfering with the actual imaging process, thus reducing acquisition time and sample exposure.
2Measurement precision
If high intensity excitation radiation is used for wavefront measurement, then measurement sensitivity is improved, but sample damage increases
Solution Approach 1:
The patent extracts the wavefront measurement function from the main imaging process by using a separate test beam. This test beam is specifically designed for wavefront measurement and can be optimized independently. The measurement beam path is separated from the imaging beam path, allowing the use of appropriate intensity levels for measurement without compromising sample integrity during actual imaging.
Solution Approach 2:
The patent changes the parameters of the test beam (such as intensity, wavelength, or pulse duration) to be optimized for wavefront measurement rather than for sample imaging. By adjusting these parameters, the system achieves sufficient measurement sensitivity while minimizing the harmful effects on the sample. The test beam parameters are independently controllable from the imaging beam parameters.
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 faster and more sensitive image acquisition with high penetration depth and reduced sample damage, capable of observing dynamic processes in radiation-sensitive samples.
Implementation Method 1
a first detection radiation caused by the action of the first excitation radiation is detected and evaluated using a wavefront detector with regard to the formation of its wavefront
Implementation Method 2
a second excitation radiation is manipulated using a wavefront manipulator in order to correct any detected deviations in the wavefront
Implementation Method 3
focusing a first excitation radiation into a volume of the sample
Implementation Method 4
The nonlinearity of the excitation ensures that when fluorescence events are excited, fluorescence emission is only emitted from the focus
Data Source
Figure 1
Figure 2
Figure 3a~3b
AI summary
The invention relates to a method and a device for imaging a sample (14). In the method, a first excitation beam (5) is focused into a volume of the sample (14), and the resulting first detection beam (15) is detected and evaluated with regard to the formation of its wavefront. Based on the evaluation results, a second excitation beam (11) is manipulated to correct any detected deviations of the wavefront. A region (20) of the sample (14) to be imaged is scanned by means of the second excitation beam (11), and a second detection beam (16) is acquired as image data. A characteristic feature is that the second excitation beam (11) is directed into the sample volume in at least two partial beams (11T), each illuminating a spot (22) within the sample volume, and the second detection beams (16) caused by each partial beam (11T) are acquired separately.