Correlative Microscopy for Protein Complex Spatial Analysis
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Solution Overview
Problem
Current methods for studying protein interactions at the nanoscale in cells are limited by low spatial resolution and inability to examine individual cells efficiently, missing crucial information for drug development, especially for cancer and AIDS research.
Innovation Solution
A device and method combining a sample processing unit with temperature control, fluid management, and a correlative scanning transmission electron microscope (STEM) and light microscope system, allowing for high-resolution imaging and automated data analysis of protein distribution in intact cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Illumination intensity
If light microscopy is used to visualize protein locations, then spatial distribution can be observed, but spatial resolution is limited and direct visualization of protein complexes is not possible
Solution Approach 1:
The patent combines light microscopy and electron microscopy into a correlative imaging system. The light microscope visualizes fluorescently labeled proteins to identify their spatial distribution, while the electron microscope provides high-resolution imaging of the same region to directly observe protein complexes. This merging of two microscopy techniques allows both low-resolution wide-field visualization and high-resolution structural detection.
Solution Approach 2:
The patent uses fluorescent tags as intermediaries to mark specific proteins. These fluorescent tags are attached to target proteins and allow the light microscope to locate them. The same fluorescent tags or associated structures then serve as reference points for the electron microscope to image the corresponding protein complexes with high spatial resolution.
2Reliability
If FRET is used to detect complex formation, then presence of neighboring molecules can be inferred, but precise spatial resolution and distinction of complex components is not possible
Solution Approach 1:
The patent merges FRET-based detection with electron microscopy imaging. FRET provides reliable detection of protein-protein interactions through energy transfer between fluorophores, while electron microscopy simultaneously or subsequently images the same region to provide high-resolution structural information about the complexes and their components.
Solution Approach 2:
The patent uses fluorescent tags as intermediaries that serve dual purposes: they enable FRET detection of complex formation and also serve as localization markers for electron microscopy imaging, thereby linking functional detection with structural visualization.
3Measurement precision
If conventional electron microscopy is used to examine thin sections, then nanoscale dimensions can be investigated, but cells are not intact and examination is very time-consuming
Solution Approach 1:
The patent performs preliminary actions by preparing cells in a state that preserves their integrity and facilitates subsequent rapid imaging. Cells are cultured and labeled with fluorescent tags before imaging, and the correlative microscopy system is pre-configured to automatically acquire both light and electron microscopy images of the same region, eliminating the need for time-consuming manual section preparation and alignment.
Solution Approach 2:
The patent merges automated image acquisition and processing workflows for both light and electron microscopy. The system automatically coordinates imaging of the same cellular region with both techniques, and software automatically aligns and correlates the images, dramatically reducing the time required compared to manual examination of multiple thin sections.
4Ease of operation
If light microscopy with fluorescent tags is used, then protein locations can be visualized, but it is not possible to directly visualize whether certain proteins form a complex or not
Solution Approach 1:
The patent merges light microscopy and electron microscopy into a correlative imaging system. The light microscope visualizes fluorescently labeled proteins to identify their spatial distribution, while the electron microscope provides high-resolution imaging of the same region to directly observe protein complexes. This merging of two microscopy techniques allows both low-resolution wide-field visualization and high-resolution structural detection.
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 precise spatial analysis of protein complexes in individual cells, enhancing our understanding of cellular functions and facilitating the development of targeted therapies by providing detailed information on protein distribution and interaction.
Implementation Method 1
an electron microscope with a STEM detector
Implementation Method 2
the light microscope comprises a light-optical lens having a cavity
Implementation Method 3
the light microscope comprises a light-optical lens having a cavity
Data Source
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AI summary
The present invention relates to a device and a method for the stoichiometric analysis of samples. In order to study the spatial distribution of different proteins in the plasma membrane of a complete cell within a short time frame, a device and a method are proposed for the stoichiometric analysis of samples. The problem is solved by a device for the stoichiometric analysis of samples, wherein the device comprises a) a sample processing device, comprising a sample holder for holding the sample, means for setting the temperature, means for adding and removing fluid (also gases) and at least one fluid reservoir, b) an electron microscope with a detector, and c) a process control device, controlled by a computer, for controlling the means for setting the temperature and the means for adding and removing fluids (also gases) and an image recording device, automated and controlled by a computer, which captures images by means of the electron microscope, a unit storing the captured images, and an image analysis unit controlled by the computer.