Laser Poration of Cells Using Thermoplasmonic Substrates
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Solution Overview
Problem
Current intracellular delivery methods lack the ability to efficiently, safely, and cost-effectively deliver diverse cargos such as amino acids, proteins, and nucleic acids into cells with high throughput, spatial selectivity, and without immunotoxicity, while maintaining cell viability.
Innovation Solution
A method involving a substrate with metalized projections coated with an electrically conductive layer, where cells are exposed to continuous or pulsed laser radiation, causing transient changes in cell permeability to facilitate cargo uptake, using thermoplasmonic substrates with pyramidal projections and controlled laser parameters for efficient and selective delivery.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If viral transduction is used for intracellular delivery, then delivery capability is achieved, but immunotoxicity risks and limited cargo-carrying capacity occur
Solution Approach 1:
The patent replaces biological viral transduction mechanisms with a physical laser-based poration system. The laser radiation directly creates pores in the cell membrane through photothermal effects, eliminating the need for viral vectors and their associated immunotoxicity while enabling delivery of diverse cargos including proteins, nucleic acids, and drugs.
Solution Approach 2:
The patent utilizes changes in optical and thermal parameters (laser wavelength, intensity, pulse duration) to control the poration process. By adjusting these parameters, the system can create transient pores that facilitate cargo entry without causing permanent damage or immunotoxicity, thereby resolving the contradiction between delivery capability and safety.
2Adaptability or versatility
If conventional laser poration is used, then cell permeability is increased for cargo delivery, but cell viability decreases
Solution Approach 1:
The patent employs pulsed laser radiation with specific pulse durations (nanosecond to microsecond range) followed by rest intervals. This periodic action allows the cell membrane to repair and stabilize between pulses, maintaining viability while accumulating sufficient poration effect for efficient cargo delivery over multiple cycles.
Solution Approach 2:
The system dynamically adjusts laser parameters (intensity, pulse width, repetition rate) based on real-time feedback and cell response. This dynamic control enables optimization of the poration process to achieve maximum cargo delivery efficiency while minimizing damage to cell structure and function, thereby preserving viability.
3Productivity
If high laser intensity is applied for rapid cargo delivery, then delivery speed increases, but substrate damage occurs
Solution Approach 1:
The patent employs substrates with localized features such as arrays of protrusions or patterns that concentrate laser energy delivery to specific regions. This local quality approach allows high-intensity laser treatment in targeted areas for rapid cargo delivery while preserving the overall substrate integrity and preventing widespread damage.
Solution Approach 2:
The patent uses pulsed laser radiation with controlled duty cycles, applying high intensity only during brief pulse windows followed by longer rest periods. This periodic application accumulates delivery throughput while allowing heat dissipation and substrate recovery between pulses, preventing thermal damage and maintaining substrate strength.
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
Achieves high efficiency and viability of cargo delivery to cells, with up to 95% efficiency and 98% viability, enabling the delivery of various cargos at a high throughput rate, including macromolecules and proteins, while minimizing immunotoxicity and substrate damage.
Implementation Method 1
irradiating the substrate surface (and in particular the projections) with continuous or pulsed radiation
Implementation Method 2
using thermoplasmonic substrates with pyramidal projections and controlled laser parameters for efficient and selective delivery
Implementation Method 3
causing transient changes in cell permeability to facilitate cargo uptake
Data Source
AI summary
In one aspect, a method of cell processing is disclosed, which includes disposing a plurality of cells on a substrate across which a plurality of projections are distributed and an electrically conductive layer at least partially coating said projections, exposing the cells to a cargo to be internalized by the cells, irradiating the substrate surface (and in particular the projections) with continuous wave or pulsed laser radiation. For example, one or more laser pulses having a pulse width in a range of about 1 ns to about 1000 ns can be applied so as to facilitate uptake of the cargo by at least a portion of the cells (e.g., the cells positioned in the vicinity of the projections (e.g., within hundreds of nanometer (such as less than 100 nm) of the projections)). In some embodiments, the laser pulses have a pulse width in a range of about 10 ns to about 500 ns, e.g., in a range of about 5 ns to about 50 ns.