Laser Ablation Imaging with Real-Time Focus Control for Live Cells
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Solution Overview
Problem
Conventional methods struggle to obtain sub-cellular biological material from living cells for effective downstream analysis due to large ablation spot sizes and the need for sample fixation, which limits access to intra- and inter-cellular material.
Innovation Solution
A device with an optical assembly and laser focus detection unit allows real-time monitoring and adjustment of ablation parameters, enabling precise ablation of sub-cellular regions with minimal impact on cellular structure, using high-NA optics and directing laser light parallel to the ablation plume for efficient transfer to a receiver.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If conventional LAESI methods are used, then biomolecules can be analyzed, but the ablation spot size is too large to target sub-cellular regions
Solution Approach 1:
The patent changes the optical parameter by using high numerical aperture (NA) optics instead of conventional low NA optics. This parameter change reduces the ablation spot size from being larger than whole cells to sub-cellular dimensions, enabling precise targeting of specific intracellular regions while maintaining effective ablation and biomolecule collection.
2Reliability
If sample fixation is performed for material collection, then material can be obtained for analysis, but live cells cannot be monitored in real-time
Solution Approach 1:
The patent extracts the requirement for sample fixation by implementing a rapid ablation and collection system that captures biomolecules directly from live cells before fixation is needed. The high-speed ablation and immediate transfer of material to the receiver allows analysis of live cell material without the compromising effects of fixation and drying.
3Productivity
If laser ablation is performed with conventional optics, then material can be removed, but the remaining cellular structure is excessively damaged
Solution Approach 1:
The patent applies local quality by using high NA optics to concentrate the laser energy into a tightly focused sub-cellular spot. This creates a localized ablation zone that removes only the targeted region while minimizing thermal and mechanical damage to the surrounding cellular structure, allowing the cell to survive and remain viable for further analysis.
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 collection of targeted biological material with minimal degradation, allowing for effective downstream analysis and survival of live cells, with improved spatial and temporal resolution.
Implementation Method 1
an ablation laser...configured to direct laser light through the objective, through the sample stage and into the sample to selectively ablate at least a portion of the sample
Implementation Method 2
laser-ablated material are ionized by nanometer-sized droplets
Implementation Method 3
The low NA optics used for ablation of the sample lead to large ablation spot sizes...capable of access to sub-cellular detail
Implementation Method 4
laser-ablated material are ionized by nanometer-sized droplets from an electrospray ion source which are then transmitted to a mass spectrometer
Data Source
AI summary
Disclosed herein are systems for imaging and ablating a sample. An imaging/ablating device includes an optical assembly, a sample stage, and a receiver. The optical assembly enables ablation of a region of interest within the sample. The laser light propagated from the optical assembly during ablation propagates substantially in the same direction as the direction of travel of the ablation plume toward the receiver. A laser focus detection unit including at least one reference laser and photodetector generates at least one real-time detection signal indicative of one or more characteristics of the sample during ablation and/or of a distance from the objective to the sample stage or a surface of the sample. A controller coupled with the laser focus detection unit dynamically controls in real-time one or more parameters of the ablation laser and/or a position of the objective and/or a position of the receiver relative to the sample to improve MS imaging quality.