Protective Agent for Vacuum Electron Microscopy of Living Samples
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Solution Overview
Problem
Current electron microscopy techniques cannot observe biological samples in their living state at high magnification without deforming or killing them, as they require a vacuum environment that is detrimental to water-containing samples and prevents oxygen supply, leading to dehydration and ice crystal formation.
Innovation Solution
A method involving a protective agent containing saccharides and electrolytes, such as glycerin and glucose, is applied to biological samples to form a thin film using an electron beam or plasma, allowing for observation in a vacuum without deformation, and a kit including a surfactant solution for electron microscopic observation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a biological sample is placed in a vacuum environment for electron microscopic observation, then the observation can be performed, but the sample undergoes dehydration and deformation
Solution Approach 1:
The patent introduces an ionic liquid as an intermediary substance between the vacuum environment and the biological sample. The ionic liquid forms a protective layer around the sample, allowing vacuum conditions to be maintained while preventing direct vacuum-induced dehydration and structural deformation of the biological specimen.
Solution Approach 2:
The patent creates an inert environment using ionic liquid that mimics a protective atmosphere around the sample. This inert environment prevents harmful vacuum effects while allowing electron microscopic observation to proceed, effectively shielding the sample from vacuum damage.
2Reliability
If a biological sample is vacuum-dried to remove moisture, then vacuum resistance is improved, but the surface shape is significantly deformed
Solution Approach 1:
The ionic liquid acts as a mediator that provides vacuum resistance without requiring vacuum drying. It forms a protective barrier that allows the sample to maintain its natural hydrated state and surface shape while still being compatible with vacuum observation conditions.
Solution Approach 2:
The patent changes the physical-chemical parameters of the sample environment by introducing ionic liquid, which has unique properties allowing it to maintain sample hydration and structure under vacuum conditions, thereby preserving surface shape while providing vacuum resistance.
3Reliability
If a conductive film is deposited on a non-conductive sample, then conductivity is improved, but the sample requires pretreatment that may alter its natural state
Solution Approach 1:
The ionic liquid serves as an intermediary that provides both conductivity and vacuum compatibility simultaneously. This eliminates the need for separate conductive coating steps and complex pretreatment procedures, simplifying sample preparation while maintaining the sample's natural state.
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 the observation of water-containing biological samples, like mammals, plant tissues, and cultured cells, in their living state without deformation, maintaining their viability and structure for imaging.
Implementation Method 1
forming a thin-film on a surface of the water-containing biological sample by irradiating the water-containing biological sample with an electrom beam
Implementation Method 2
forming a thin-film on a surface of the water-containing biological sample by irradiating the water-containing biological sample with an electrom beam or plasma
Data Source
Figure 1(a)~1(b)
Figure 2(a)~2(b)
Figure 3(a)~3(b)
AI summary
Provided are a protective agent for electron microscopic observation in a vacuum which can protect a biological sample in a water-containing state, such as a mammal, a plant tissue or a cultured cell, and a single cell in the living state without deforming it even in a vacuum, a kit using the same, methods for observation, diagnosis, evaluation, and quantification of a sample by an electron microscope, and a sample stage to be used for the observation. The protective agent for electron microscopic observation of the present invention contains a survival environment imparting component, a saccharide, and an electrolyte.