Combination Microscopy Resolving Resolution-Speed Trade-offs

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Solution Overview

Problem

High-resolution microscopy methods face limitations in measuring speed, sample field size, and penetration depth, making them unsuitable for examining large samples or dynamic processes, especially when high spatial resolution is required.

Innovation Solution

A combination microscope that integrates multiple microscopy methods, including structured line or wide-field illumination, activation of marking molecules, laser scanning microscopy, and STED techniques, to generate composite images with enhanced spatial resolution, allowing for high-resolution imaging of small sample sections and dynamic processes.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If high-resolution microscopy methods are used, then spatial resolution is improved, but measuring speed deteriorates

Engineering Contradiction:
Improvespatial resolutionVSAvoidmeasuring speed
Core Design Contradiction:
Measurement precisionVSProductivity

Solution Approach 1:

The patent divides the sample field into multiple regions of interest (ROIs) and applies high-resolution microscopy only to selected ROI regions while using classic microscopy for other areas. This segmentation allows the system to achieve high spatial resolution where needed while maintaining faster measurement speeds for less critical areas, thus resolving the contradiction between resolution and measuring speed.

Inventive Principle:
Principle #1Segmentation

2Measurement precision

If high-resolution microscopy methods are used, then spatial resolution is improved, but the measurable sample field size deteriorates

Engineering Contradiction:
Improvespatial resolutionVSAvoidsample field size
Core Design Contradiction:
Measurement precisionVSArea of stationary object

Solution Approach 1:

The patent implements local quality by applying high-resolution microscopy methods selectively to specific regions of interest within the sample field, while using classic microscopy methods for the remaining areas. This allows the system to provide enhanced spatial resolution locally where it is most needed, while maintaining a larger overall measurable sample field size through the use of faster classic microscopy for other regions.

Inventive Principle:
Principle #3Local quality

3Measurement precision

If high-resolution microscopy methods are used, then spatial resolution is improved, but penetration depth deteriorates

Engineering Contradiction:
Improvespatial resolutionVSAvoidpenetration depth
Core Design Contradiction:
Measurement precisionVSLength of stationary object

Solution Approach 1:

The patent implements a dynamic switching mechanism that allows the microscopy system to adaptively choose between high-resolution methods and classic methods based on the specific imaging requirements. When deep penetration is needed, the system can switch to classic microscopy methods that offer better penetration depth, while maintaining the option to use high-resolution methods for superficial structures, thus dynamically resolving the contradiction between resolution and penetration depth.

Inventive Principle:
Principle #15Dynamics

4Reliability

If multiple microscopy methods are combined, then data quality is improved, but device complexity increases

Engineering Contradiction:
Improvedata qualityVSAvoiddevice complexity
Core Design Contradiction:
ReliabilityVSDevice complexity

Solution Approach 1:

The patent implements multi-functionality by designing a microscopy system that can perform multiple microscopy methods (high-resolution and classic) using a unified platform. The system includes a beam combining unit that can selectively direct different illumination paths and a detector that can process multiple types of microscopy signals, allowing one device to serve multiple functions and reduce overall system complexity despite combining multiple methods.

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

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-resolution imaging of small sample sections while maintaining classic microscope data quality, simplifying measuring processes and increasing data quality, and allows for simultaneous recording of different image fields with varying resolutions, overcoming the limitations of traditional high-resolution microscopy.

Implementation Method 1

an illumination radiation modulator arranged downstream of the wide-field illumination module in the illumination beam path in the illumination direction, which can be activated and deactivated in the illumination beam path and in the activated state imposes a strip-shaped modulation on the illumination radiation

Methodology Applied
Scientific EffectStructured illumination:

Implementation Method 2

an objective, which detects the sample for all microscopy methods

Methodology Applied
Scientific EffectLuminescence detection: Luminescence

Implementation Method 3

a detection beam path and a microscope module connected to the detection beam path, which has a tube lens and a surface detector and, together with the lens, images the sample onto the surface detector

Methodology Applied
Scientific EffectOptical imaging: Lens

Data Source

PatentEP3206070B1Combination microscopy
Publication Date: 2018.05.23 CARL ZEISS MICROSCOPY GMBH
  • EP3206070B1 patent drawingFigure 1~2
  • EP3206070B1 patent drawingFigure 3
  • EP3206070B1 patent drawingFigure 4

AI summary

A method for generating an image of a sample using microscopy techniques with varying spatial resolutions is described, combining at least two of the following microscopy techniques: laser scanning microscopy to generate an LSM microscopy image; a microscopy technique in which the sample is excited to luminescence by structured line or wide-field illumination, the structure is rotated and shifted multiple times for each rotation, with at least three rotation positions and at least three shift positions per rotation position; the luminescent sample is imaged onto an area detector in each case, and a first microscopy image with increased spatial resolution beyond the optical resolution of the image is generated from the images thus obtained; and a further microscopy technique according to the PAL principle, with which a second microscopy image is generated.specify the geometric locations of luminescence-emitting labeling molecules with a spatial resolution increased beyond the optical resolution, and yet another microscopy method in which the sample is labeled with labeling molecules suitable for STED, ESA or RESOLFT techniques and a third microscopy image is generated using STED, ESA or RESOLFT, whereby the obtained microscopy images are superimposed.