Light Input Assembly for Targeted Illumination in Confocal Microscopy

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Solution Overview

Problem

Scanning confocal microscopy systems, particularly those using spinning disk configurations, lack the ability to increase light intensity dramatically over a specified region, limiting their effectiveness in targeted illumination applications such as FRAP and photoactivation experiments.

Innovation Solution

A light input assembly is inserted between the confocal scanning head and the microscope, featuring a light input, beam directing means, a beam coupler, and an optical relay to selectively illuminate and modify specific regions of a sample, allowing for controlled and intense light delivery without compromising the operation of the confocal scanning head.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If a spinning disk configuration with multiple pinholes is used for parallel illumination, then imaging speed is improved, but the ability to increase light intensity dramatically over a specified region deteriorates

Engineering Contradiction:
Improveimaging speedVSAvoidlight intensity over selected region
Core Design Contradiction:
ProductivityVSIllumination intensity

Solution Approach 1:

The system separates the illumination function into two independent paths: a spinning disk confocal path for high-speed imaging and a separate targeted illumination path for high-intensity regional illumination. This segmentation allows each path to be optimized for its specific function without compromising the other.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

A beam coupler is introduced as an intermediary component that selectively combines the confocal illumination beam with the targeted illumination beam. The beam coupler allows dynamic switching between confocal imaging mode and targeted illumination mode, enabling both functions to coexist in the same optical path.

Inventive Principle:
Principle #24Intermediary (Mediator)

2Illumination intensity

If laser power is increased for targeted illumination in scanning spot systems, then illumination intensity is improved, but imaging speed deteriorates

Engineering Contradiction:
Improvelight intensityVSAvoidimaging speed
Core Design Contradiction:
Illumination intensityVSProductivity

Solution Approach 1:

The illumination system is divided into two independent illumination paths: a confocal scanning path for high-speed imaging and a separate targeted illumination path for high-power regional illumination. This allows simultaneous operation of both functions without the trade-off present in single-path systems.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The patent merges two previously separate illumination approaches (confocal scanning and targeted high-power illumination) into a single integrated system. The beam coupler enables both illumination paths to be combined and directed to the sample simultaneously or alternately, eliminating the need to choose between speed and intensity.

Inventive Principle:
Principle #5Merging (Combining)

3Adaptability or versatility

If an assembly is inserted between the confocal scanning head and microscope for targeted illumination, then illumination versatility is improved, but system complexity deteriorates

Engineering Contradiction:
Improveillumination versatilityVSAvoidsystem complexity
Core Design Contradiction:
Adaptability or versatilityVSDevice complexity

Solution Approach 1:

The inserted assembly is designed as a multi-functional unit that can perform both confocal imaging and targeted illumination functions. By integrating multiple functions into a single assembly, the patent reduces the need for separate independent systems while maintaining versatility.

Inventive Principle:
Principle #6Universality (Multi-functionality)

Solution Approach 2:

The assembly acts as an intermediary between the confocal scanning head and the microscope, providing targeted illumination capabilities without requiring modification of the existing confocal system. This intermediary approach allows addition of functionality while preserving the original system's operation.

Inventive Principle:
Principle #24Intermediary (Mediator)

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-power, targeted illumination of selected regions within the sample, facilitating FRAP and photoactivation experiments with improved precision and efficiency, while maintaining the functionality of the confocal scanning head.

Implementation Method 1

a beam coupler for selectively coupling the light beam into the light path from the confocal scanning head to the microscope

Methodology Applied
Scientific EffectOptical coupling:

Implementation Method 2

an optical relay on said light path within the assembly to relay a sample image formed on the light path between the confocal scanning head and the microscope from the microscope side of the assembly to the scanning confocal head side of the assembly

Methodology Applied
Scientific EffectOptical relay:

Implementation Method 3

the intensity of the light beam may be sufficient to bleach or otherwise modify the optical properties of fluorescent parts of the selected region of the sample, to enable FRAP or photoactivation experiments

Methodology Applied
Scientific EffectPhotobleaching:

Implementation Method 4

many of the dyes used in fluorescence microscopy will 'bleach' when exposed to very strong light sources

Methodology Applied
Scientific EffectFluorescence: Fluorescence

Data Source

PatentEP2059844B1Improvements in and relating to scanning confocal microscopy
Publication Date: 2017.05.17 REVVITY SINGAPORE PTE LTD
  • EP2059844B1 patent drawingFigure 1
  • EP2059844B1 patent drawingFigure 2~3
  • EP2059844B1 patent drawingFigure 4~6

AI summary

An assembly (100) and a method are provided for inputting a light beam into a light path extending from the confocal scanning head (102) to the microscope (104) of a scanning confocal microscope system to illuminate a selected region of a sample mounted in the microscope. The assembly comprises a light input for receiving a light beam from a light source; beam directing means (8, 12) for controlling the path of the light beam with reference to the shape of the selected region of the sample; and a beam coupler (16) for selective coupling the light beam into the light path from the confocal scanning head (102) to the microscope (104), with the beam direction being controlled by the beam directing means so as to illuminate the selected region. A scanning confocal microscope system including such an assembly is also described, together with a method of calibrating the system. In addition, an optical switch for selectively switching a light beam between two paths is disclosed.