Light-field Microscope Illumination Using Perpendicular Beam Width Control
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Solution Overview
Problem
Conventional light-field microscopes face challenges in efficiently constructing three-dimensional image data while preventing sample fading, as they often irradiate excitation light beyond the necessary areas, leading to unnecessary exposure and potential fading of the sample.
Innovation Solution
The light-field microscope incorporates an illumination optical system that radiates excitation light with a predetermined width perpendicular to the optical axis of the objective lens, using a microlens array to collect and focus fluorescence, allowing for efficient acquisition of three-dimensional image data by limiting excitation light to the necessary area and preventing sample fading. This system includes a beam-width adjuster and a scanner for precise control of the excitation light, ensuring that only the required area is irradiated for image construction.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If excitation light is radiated broadly to capture deep parts of the sample, then three-dimensional image data can be constructed, but unnecessary areas are irradiated causing sample fading
Solution Approach 1:
The illumination optical system is designed to radiate excitation light with a specific beam width that corresponds precisely to the detection area defined by the objective lens focal plane and microlens array. This ensures that excitation light is applied only to the local region where fluorescence detection is occurring, preventing unnecessary irradiation of other sample areas and thus avoiding sample fading while still enabling three-dimensional image data acquisition.
Solution Approach 2:
The system uses a beam width that is precisely controlled to match the detection field of view, avoiding excessive illumination. By setting the beam width to correspond to the area captured by the microlens array and objective lens combination, the system applies just enough excitation light for the required depth imaging without over-irradiating surrounding areas.
2Object-affected harmful factors
If a beam with predetermined width is radiated perpendicular to the optical axis, then unnecessary area irradiation is prevented, but the complexity of the illumination optical system increases
Solution Approach 1:
The illumination optical system controls the beam width parameter of the excitation light to match the detection area. By adjusting and maintaining the beam width at a specific value that corresponds to the focal plane area captured by the objective lens and microlens array, the system prevents sample fading without requiring complex additional components beyond standard optical elements.
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
This configuration enables the efficient acquisition of three-dimensional image data by preventing sample fading and allowing for the construction of images with larger depths without irradiating unnecessary areas, maintaining the sample's integrity throughout the imaging process.
Implementation Method 1
an objective lens that collects fluorescence generated in the sample as a result of the sample being irradiated with the excitation light
Data Source
AI summary
The present invention provides a light-field microscope including: an illumination optical system that radiates excitation light onto a sample; and a detection optical system including an objective lens that collects fluorescence generated in the sample as a result of the sample being irradiated with the excitation light by the illumination optical system, an image-acquisition element that acquires an image of the fluorescence collected by the objective lens, and a microlens array disposed between the image-acquisition element and the objective lens. The illumination optical system radiates a beam of the excitation light having a predetermined width in the optical-axis direction of the objective lens so as to include the focal plane of the objective lens onto the sample in a direction substantially perpendicular to the optical axis.