Biological Ion Soft-Landing for Native-State Charged Particle Imaging
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Solution Overview
Problem
Current structural imaging techniques for biological macromolecules, such as proteins and DNA, are limited by sample size and conformation, with proteins needing crystallization, which forces them into unstable conformations and restricts the types of proteins that can be studied, while existing integrated sample preparation and imaging systems lack flexibility and integration.
Innovation Solution
A monolithic system integrating sample preparation, ion transport, and imaging using a quadrupole mass filter, energy reduction cell, and electron transparent substrates for soft-landing and holographic imaging with low-energy charged particles, enabling the analysis of proteins and complexes in their native-like state without crystallization, and capable of imaging smaller proteins and non-crystallizable samples.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If proteins are crystallized for imaging, then structural images can be obtained, but the proteins are forced into unstable conformations and not all proteins can be prepared
Solution Approach 1:
The patent changes the physical state parameter of the sample from crystalline to gas-phase ion, eliminating the need for crystallization. This allows proteins and complexes in various conformations to be imaged without being forced into crystal structures, thereby maintaining native-like states while achieving high-resolution imaging
Solution Approach 2:
The patent replaces the mechanical crystallization process with an electromagnetic ionization and manipulation system. By ionizing samples and using electric fields to guide and focus ions onto substrates, the system eliminates the mechanical constraints of crystallization while achieving precise sample positioning for imaging
2Manufacturing precision
If crystallization is used for protein preparation, then imaging can be performed, but the conformations obtained may not be stable or native-like
Solution Approach 1:
The patent changes the preparation approach from solid-state crystallization to gas-phase ion deposition. This parameter change preserves the native conformation of proteins by avoiding the structural constraints of crystal lattices, while the automated ion guidance system simplifies the preparation process
Solution Approach 2:
The patent introduces an electron transparent substrate as an intermediary between the ion source and the imaging system. This substrate receives ion deposits and positions them for imaging without requiring crystallization, thereby preserving native conformations while enabling high-resolution imaging
3Measurement precision
If current imaging techniques are used, then structural information can be obtained for large proteins, but smaller proteins and non-crystallizable samples cannot be studied
Solution Approach 1:
The patent creates a universal imaging system that handles samples across a wide size range by ionizing them. The electromagnetic manipulation system can focus ions of various masses onto the substrate, enabling imaging of small proteins, large complexes, and non-crystallizable samples with a single methodology
Solution Approach 2:
The patent replaces size-dependent mechanical crystallization with electromagnetic ion manipulation. This substitution allows precise control of ion trajectories regardless of sample mass, enabling imaging of samples from small proteins to large complexes that would be impossible to crystallize
4Ease of operation
If integrated sample preparation and imaging systems are used, then workflow is simplified, but flexibility in sample type and size is reduced
Solution Approach 1:
The integrated system incorporates universal ionization and electromagnetic manipulation components that can handle diverse sample types. The system's core functionality—ion generation, guidance, and deposition—applies equally to proteins, complexes, and various conformations, maintaining flexibility while providing workflow integration
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 imaging of proteins and complexes in their native-like state, overcoming size and conformation limitations, and providing higher resolution and flexibility in sample type and size, with reduced damage and noise, compared to existing methods like cryo-EM and X-ray crystallography.
Implementation Method 1
an ion filter coupled to select a sample ion from an ionized sample supply, the ion filter including a quadrupole filter to select the sample ion from the sample supply
Implementation Method 2
an energy reduction cell coupled to receive the selected sample ion and reduce a kinetic energy of the sample ion
Implementation Method 3
an ion transport module coupled to receive the sample ion from the ion filter and transport the sample ion to the substrate
Implementation Method 4
a charge particle emitter coupled to direct coherent charged particles toward the sample; and a detector arranged to detect interference patterns formed from interaction of the coherent charged particles and the sample
Implementation Method 5
a detector arranged to detect interference patterns formed from interaction of the coherent charged particles and the sample
Data Source
AI summary
Systems and method for the preparation and delivery of biological samples for charged particle analysis are disclosed herein. An example system at least includes an ion filter coupled to select a sample ion from an ionized sample supply, the ion filter including a quadrupole filter to select the sample ion from the sample supply, an energy reduction cell coupled to receive the selected sample ion and reduce a kinetic energy of the sample ion, a validation unit coupled to receive the sample ion and determine whether the sample ion is a target sample ion, a substrate coupled to receive the sample, wherein the substrate is electron transparent, an ion transport module coupled to receive the sample ion from the ion filter and transport the sample ion to the substrate, and an imaging system arranged to image, with a low energy charged particle beam, the sample located on the substrate, wherein the substrate is arranged in an analysis location. The imaging system including a charge particle emitter coupled to direct coherent charged particles toward the sample; and a detector arranged to detect interference patterns formed from interaction of the coherent charged particles and the sample.