Remote Focusing Microscope Polarization Splitting
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Solution Overview
Problem
Current microscopy techniques face challenges in achieving fast and efficient 3D volumetric imaging, particularly with fragile samples, due to limitations in sample scanning and optical sectioning methods, which result in vibration artifacts, reduced scan speed, and light loss.
Innovation Solution
A remote focusing system that splits the fluorescence signal into polarized light beams, allowing for aberration-free, multi-color, volumetric imaging without compromising the fluorescent signal or requiring sample/detection-objective translation. This system includes a polarizing beam splitter, remote objective, and mirrors to direct and combine the polarized light beams at a detection surface.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If sample scanning is used to perform 3D volumetric imaging, then images at different planes can be obtained, but vibration artifacts occur and scan speed is reduced
Solution Approach 1:
Instead of moving the sample or detection objective to change focus planes, the patent inverts the approach by using a remote focusing system that moves the focal plane while keeping both the sample and objective stationary. This is achieved by splitting the fluorescence signal into polarized light beams and using a movable mirror to adjust the focal position, thereby eliminating vibration artifacts while maintaining high scan speed.
2Measurement precision
If detection objective is moved to perform optical sectioning, then light sheet synchronization can be achieved, but moving optical parts through media is required and imaging speed is reduced
Solution Approach 1:
The patent replaces the mechanical movement of the detection objective with an optical-based remote focusing system. By using polarizing beam splitters and a movable mirror to adjust the focal plane, the system achieves optical sectioning without physically moving the objective through the sample media, thereby eliminating mechanical constraints and improving imaging speed.
3Measurement precision
If remote focusing with polarization splitting is used, then aberration-free volumetric imaging is achieved, but fluorescent signal may be compromised
Solution Approach 1:
The patent segments the fluorescence signal into two separate polarized light beams using a polarizing beam splitter. Each polarized beam is then directed through a separate optical path with its own objective lens, allowing both beams to be imaged simultaneously without interference. This segmentation enables the system to maintain full fluorescent signal intensity while achieving aberration-free volumetric imaging through the remote focusing mechanism.
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 high-speed 3D volumetric imaging with minimal light loss, maintaining image quality and allowing for simultaneous multi-color imaging, which is not feasible with existing pmRF systems that incur up to 50% light loss.
Implementation Method 1
a polarizing beam splitter configured to split an unpolarized light beam that is incident upon the beam splitter into a first polarized light beam having a first polarization and a second polarized light beam having a second polarization
Implementation Method 2
a remote objective configured to focus the first polarized light beam and the second polarized light beam onto a focal plane
Implementation Method 3
a first mirror configured to direct the first polarized light beam to the remote objective, and a second mirror configured to direct the second polarized light beam to the remote objective
Data Source
AI summary
Various embodiments of a remote focusing system and a microscope that includes such system are disclosed. The system includes a first mirror configured to direct first and second polarized light beams from a beamsplitter to a remote objective. The first and second polarized light beams are representative of an image of a portion of a biological sample. The remote objective is configured to provide first and second polarized intermediate images of the portion of the sample based on the first and second polarized light beams to a reflective surface of a focal plane mirror. The focal plane mirror and the first and second mirrors are further configured to direct the first polarized intermediate image and the second polarized intermediate image to a detector to detect the first polarized intermediate image and the second polarized intermediate image as a final composite image.