Laser Microdissection Using Charged Membrane and Electrostatic Collection
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Solution Overview
Problem
Existing laser microdissection techniques face challenges such as mechanical damage, low precision, and high cost due to mechanical methods, UV laser alteration of DNA and RNA, and high operational complexity in collecting target regions from tissue samples.
Innovation Solution
A method and system utilizing a negatively-charged membrane on a substrate with a laser beam passing through to dissect and electrostatically propel target regions to a positively-charged agar gel collection material, avoiding mechanical contact and minimizing sample damage.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If laser catapulting technique is used to dissect target regions, then dissection precision is improved, but DNA and RNA may be altered by the UV laser beam striking the middle of the cells
Solution Approach 1:
The patent extracts the harmful UV laser beam from the collection process by using a visible light laser instead. The collection mechanism is separated from the dissection mechanism, allowing precise dissection without the harmful effects of UV radiation on the collected cells.
Solution Approach 2:
The patent introduces a charged membrane as an intermediary between the laser dissection system and the collection system. The membrane is charged to attract the dissected target regions without requiring UV laser exposure, thereby mediating the collection process without harmful radiation.
2Reliability
If laser pressure catapulting technique is used, then target regions can be collected without mechanical contact, but additional time is required to defocus and re-focus the laser
Solution Approach 1:
The patent applies preliminary action by pre-charging the membrane before the laser dissection process. This eliminates the need for laser refocusing during collection, as the charged membrane passively attracts the dissected target regions immediately after dissection, saving time.
3Productivity
If laser capture microdissection technique using thermoplastic film is used, then target regions can be collected, but mechanical damage occurs on biological samples during separation
Solution Approach 1:
The patent replaces the mechanical film-based collection system with an electrostatic field-based system. The charged membrane creates an electric field that attracts and collects the laser-dissected target regions without mechanical contact, eliminating mechanical damage while maintaining collection efficiency.
4Manufacturing precision
If sticky membrane transfer technique is used to collect dissected target regions, then collection precision is improved, but the membrane used leads to higher cost
Solution Approach 1:
The patent changes the parameter of the collection membrane from requiring special sticky properties to requiring only electrostatic charge properties. This allows the use of simpler, less expensive materials that can be charged, thereby reducing cost while maintaining collection precision through electrostatic attraction.
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
This approach enables precise, non-contact collection of target regions with reduced sample damage and operational complexity, improving efficiency and precision while maintaining bioactivity for subsequent analysis.
Implementation Method 1
passing a laser beam from a second side of the substrate, through the substrate, the membrane, and the target object, to dissect target regions from the prepared tissue section
Implementation Method 2
mounting a negatively-charged membrane on a first side of a substrate... whereby the dissected target regions adhere to the collection material
Data Source
AI summary
A method of collecting target regions from a target object is described. The method in one embodiment comprises mounting a negatively-charged membrane on a first side of a substrate, mounting a target object on the membrane, positioning a collection material adjacent to the target object, and passing a laser beam from a second side of the substrate, through the substrate, the membrane, and the target object, to dissect target regions from the prepared tissue section, whereby the dissected target regions adhere to the collection material. In another embodiment, the present invention is a system for collecting target regions from a target object. In one embodiment, the system comprises a substrate having a first side and a second side, a negatively-charged membrane adhered to the first side of the substrate, and a collection material mountable adjacent to the membrane. In another embodiment, the system further comprises an inverted microscope, a stage for holding the substrate over the microscope, a generator operable to generate a laser beam to pass through the substrate from the second side and to dissect target regions from a target object mounted on the membrane, whereby the dissected target regions adhere to the collection material. In the preferred embodiments, the target objects are tissues and the target regions are cells.