Diamond Reference Samples for Non-Bleaching Microscope Calibration

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Solution Overview

Problem

Existing reference samples for calibrating optical imaging systems, particularly high-resolution microscopes, are limited in their ability to accurately measure and adjust resolution due to degradation of luminescence intensity and require separate recordings of illumination and detection beam paths, which complicates direct verification of resolution capability.

Innovation Solution

A reference sample using diamond or silicon carbide as a carrier material, doped with nitrogen vacancy pairs or silicon vacancy pairs, featuring precise two- and three-dimensional structures that can be excited to emit luminescence, allowing for direct resolution measurement and calibration without degradation, and enabling comparison of different optical systems.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If glass reference samples doped with rare earth elements are used, then fluorescence radiation can be excited for resolution evaluation, but luminescence intensity decreases significantly upon repeated excitation

Engineering Contradiction:
Improveresolution measurement capabilityVSAvoidluminescence intensity stability
Core Design Contradiction:
Measurement precisionVSReliability

Solution Approach 1:

The patent changes the material parameter from glass doped with rare earth elements to diamond or silicon carbide doped with nitrogen vacancy pairs or silicon vacancy pairs. This material substitution fundamentally alters the luminescence properties, providing a reference sample that maintains stable luminescence intensity over repeated excitations while still enabling fluorescence radiation excitation for resolution measurements

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent uses composite material structures where diamond or silicon carbide serves as the base material and nitrogen vacancy pairs or silicon vacancy pairs serve as the dopant. This composite approach combines the structural stability of diamond/silicon carbide with the luminescence properties of the vacancy pairs, achieving both measurement precision and reliability

Inventive Principle:
Principle #40Composite materials

2Ease of manufacture

If technical processes with resolution corresponding to optical system resolution are used to introduce structures, then structures can be created in the substrate, but metrological determination of high-resolution microscope resolution is limited

Engineering Contradiction:
Improvestructure introduction capabilityVSAvoidresolution measurement accuracy
Core Design Contradiction:
Ease of manufactureVSMeasurement precision

Solution Approach 1:

The patent changes the resolution parameter of the structuring process by using lithographic methods that achieve lateral resolutions and positioning accuracies of 5 nm, which is significantly higher than conventional technical processes. This enables the creation of substructures with precisely known dimensions that can serve as accurate references for measuring microscope resolution capabilities

Inventive Principle:
Principle #35Parameter changes

3Ease of operation

If separate recordings of illumination and detection beam paths are required, then calibration can be performed, but direct verification of resolution capability is complicated

Engineering Contradiction:
Improvecalibration capabilityVSAvoidmeasurement process complexity
Core Design Contradiction:
Ease of operationVSDevice complexity

Solution Approach 1:

The patent merges the illumination and detection functions into a single measurement process by using the reference sample with luminescent substructures. The excited luminescence from the substructures provides direct information about the detection beam path while the excitation process characterizes the illumination, eliminating the need for separate recordings and simplifying the calibration procedure

Inventive Principle:
Principle #5Merging (Combining)

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

The solution provides robust, non-bleaching reference samples that enable precise determination of resolution and calibration of optical systems, allowing for direct verification of resolution capability and improved comparability between systems, with high reproducibility and reduced uncertainty due to luminescence degradation.

Implementation Method 1

at least one support structure made of a support material that can be excited to emit light (luminescence)

Methodology Applied
Scientific EffectLuminescence: Luminescence

Implementation Method 2

The excitation and emission of fluorescence radiation are based on doping the glass, particularly with so-called rare earth elements

Methodology Applied
Scientific EffectFluorescence: Fluorescence

Data Source

PatentEP4103995B1Reference sample for a microscope and methods for calibration and referencing
Publication Date: 2025.10.08 CARL ZEISS MICROSCOPY GMBH
  • EP4103995B1 patent drawingFigure 1
  • EP4103995B1 patent drawingFigure 2
  • EP4103995B1 patent drawingFigure 3a~3c

AI summary

The invention relates to a reference sample (1) for calibrating and/or adjusting a microscope (9) and uses of the reference sample (1). The latter has at least one carrier structure (2) made of a carrier material (3), which is excitable to emit luminescence light at least in regions of its extent, and at least one two-dimensional and/or three-dimensional structuring (5, 6, 7, 8, 14) consisting of a number of partial structures (5.1, 5.1ax, 6.1, 7.1, 8.1, 14.1). According to the invention, the carrier material (3) is diamond or silicon carbide and is doped in the regions or around the regions of the two-dimensional and/or three-dimensional structuring (5, 6, 7, 8, 14) in order to be excitable to emit the luminescence light.