Apparatus, substrate, and method for intercellular delivery of payloads by photoporation

Abstract

An apparatus for payload delivery to a cell includes a laser source configured to generate laser energy and a substrate implanted with photoacoustic launch pads. The apparatus is configured to create a pore in a cell membrane by: absorbing the laser energy by the substrate; increasing the temperature of the photoacoustic launch pads in response to the laser energy; creating a vapor nanobubble by causing fluid surrounding the cell to absorb heat and expand in response to the increased temperature of the photoacoustic launch pads; causing the vapor nanobubble to collapse, disrupting a membrane of the cell; forming a pore in the cell membrane in response to disrupting the membrane of the cell; and delivering a payload into the cell based on causing fluid surrounding the cell containing the payload to enter the cell via the pore.

Description

APPARATUS, SUBSTRATE, AND METHOD FOR INTERCELLULAR DELIVERY OF PAYLOADS BY PHOTOPORATIONCROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of and priority to U.S. Provisional Patent Application Serial No. 63 / 575,963, filed on April 8, 2024, and U.S. Provisional Patent Application Serial No. 63 / 606,349, filed on December 5, 2023, the entire contents of which are incorporated by reference herein.TECHNICAL FIELD

[0002] The present application relates to apparatus and methods for medical applications of laser-driven microfluid pumps and, in particular, to membrane permeabilization-based cell delivery.BACKGROUND

[0003] Delivering payloads to cells is a process in biotechnology and medicine, that enables the targeted delivery of therapeutic agents, genetic material, or diagnostic tools directly into specific cells. This can be achieved through various methods, including viral vectors, lipid nanoparticles, electroporation, and cell-penetrating peptides. Each method employs unique mechanisms to cross the cell membrane while preserving the integrity and functionality of the payload. The efficiency of delivery depends on factors such as the cell type, payload size, and delivery environment.SUMMARY

[0004] In accordance with aspects of the present disclosure, an apparatus for payload delivery to a cell is presented. The apparatus includes a container for holding a fluid containing the payload and the cell to which the payload is to be delivered; a laser sourceconfigured to generate laser energy; and a substrate implanted with photoacoustic launch pads, the photoacoustic launch pads configured to absorb laser energy generated by the laser source.

[0005] In an aspect of the present disclosure, a temperature of the photoacoustic launch pads may be increased based on the absorbed laser energy to create a vapor nanobubble in the fluid.

[0006] In an aspect of the present disclosure, the vapor nanobubble may create a pore in a cell membrane of the cell thereby enabling the payload contained in the fluid to enter the cell.

[0007] In an aspect of the present disclosure, the photoacoustic launch pads may be configured to: induce a nano-jet of streaming fluid, propagating from the photoacoustic launch pads towards the cell, in response to the photoacoustic launch pads absorbing the laser energy; and cause a disruption of a cell membrane by the nano-jet of streaming fluid.

[0008] In an aspect of the present disclosure, the substrate may be at least one of quartz or glass.

[0009] In an aspect of the present disclosure, the photoacoustic launch pads may include a nanopattern of metal ions on the substrate.

[0010] In accordance with aspects of the present disclosure, the photoacoustic launch pads may include a nanopattern of metal ions.

[0011] In an aspect of the present disclosure, the metal ions may include gold ions.

[0012] In accordance with aspects of the present disclosure, the container may be a microfluidic channel through which cells flow in single file.[00131 In an aspect of the present disclosure, the substrate may be part of a wall of the microfluidic channel.

[0014] In an aspect of the present disclosure, the laser source may be configured to generate pulsed laser energy.

[0015] In an aspect of the present disclosure, a method for payload delivery to a cell is presented. The method includes: applying laser energy to a substrate to heat photoacoustic launch pads implanted within the substrate. The substrate is proximate to a fluid containing the cell, and the photoacoustic launch pads are configured to absorb laser energy; and in response to the photoacoustic launch pads absorbing the applied laser energy, creating a pore in a cell membrane of a cell contained in the fluid.

[0016] In an aspect of the present disclosure, the method may further include increasing a temperature of the photoacoustic launch pads in response to absorption of the laser energy and creating a vapor nanobubble by causing fluid surrounding the cell to absorb heat and expand in response to the increased temperature of the photoacoustic launch pads.

[0017] In an aspect of the present disclosure, the method may further include causing the vapor nanobubble to collapse, disrupting a membrane of the cell.

[0018] In an aspect of the present disclosure, the method may further include forming a pore in the cell membrane in response to disrupting the membrane of the cell.

[0019] In an aspect of the present disclosure, the method may further include delivering a payload into the cell based on causing fluid surrounding the cell containing the payload to enter the cell via the pore.[0020J In an aspect of the present disclosure, the method may further include: inducing nano-jet of streaming fluid, propagating from the photoacoustic launch pads towards the cell, in response to the photoacoustic launch pads absorbing the energy from the laser energy; and disrupting a cell membrane by nano-jets of streaming fluid.

[0021] In accordance with aspects of the present disclosure, a substrate is implanted with photoacoustic launch pads. The photoacoustic launch pads are configured to increase in temperature in response to laser energy. The photoacoustic launch pads, when disposed proximate to a fluid, are further configured to induce a nano-jet of streaming fluid in response to the photoacoustic launch pads absorbing the energy from the laser energy.

[0022] In an aspect of the present disclosure, the photoacoustic launch pads may be configured to receive laser energy of a wavelength in a range of about 180nm to about 1mm.

[0023] In an aspect of the present disclosure, the photoacoustic launch pads may include a nanopattem of gold ions, and the substrate is at least one of quartz or glass.

[0024] Further details and aspects of exemplary embodiments of the present disclosure are described in more detail below with reference to the appended figures.BRIEF DESCRIPTION OF THE DRAWINGS[0025| A better understanding of the features and advantages of the disclosed technology will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the technology are utilized, and the accompanying drawings of which:

[0026] FTG. 1 depicts a front view of an apparatus for membrane permeabilization- based cell delivery, in accordance with aspects of the present disclosure;

[0027] FIG. 2 depicts the apparatus of FIG. 1 and a mammalian cell, in accordance with aspects of the present disclosure;|0028] FIG. 3 depicts a top view of the apparatus of FIG. 1, with a laser being applied to gold (Au) implanted regions of the apparatus, in accordance with aspects of the present disclosure;

[0029] FIG. 4 depicts the creation of a vapor nanobubble (VNB) by a laser pulse, in accordance with aspects of the present disclosure;

[0030] FIGS. 5 and 6 illustrate the collapsing of the VNB and the creation of a pore in the membrane of the cell, in accordance with aspects of the present disclosure;

[0031] FIG. 7 is an illustration of the delivery of substances into the cell via the newly created pore, in accordance with aspects of the present disclosure; and

[0032] FIG. 8 is a flow diagram for a method for effecting payload delivery to a cell, in accordance with aspects of the present disclosure.

[0033] Further details and aspects of various embodiments of the present disclosure are described in more detail below with reference to the appended figures.DETAILED DESCRIPTION

[0034] This disclosure relates to apparatus and methods for medical applications of a laser-driven photoacoustic microfluid pump.

[0035] Although the present disclosure will be described in terms of specific embodiments, it will be readily apparent to those skilled in this art that various modifications, rearrangements, and substitutions may be made without departing from the spirit of thepresent disclosure. The scope of the present disclosure is defined by the claims appended hereto.10036] For purposes of promoting an understanding of the principles of the present disclosure, reference will now be made to exemplary embodiments illustrated in the drawings, and specific language will be used to describe the same. It will nevertheless be understood that no limitation of the scope of the present disclosure is thereby intended. Any alterations and further modifications of the inventive features illustrated herein, and any additional applications of the principles of the present disclosure as illustrated herein, which would occur to one skilled in the relevant art and having possession of this disclosure, are to be considered within the scope of the present disclosure.|0037] The process of converting (or transforming) one form of energy into another is often referred to as transduction. A transducer is a device that is typically employed to perform such a function, and transducers can be characterized by the direction in which the physical system (e.g., pressure, temperature, sound waves, etc.) passes through them. For example, a sensor is a type of transducer that receives and responds to a signal / stimulus from a physical system (e.g., temperature) and produces an electrical signal that represents information about the physical system. An actuator, on the other hand, is a transducer that controls / generates a physical system (e.g., sound waves), in response to some electrical signal. For example, a speaker transforms an electrical signal of a recording to mechanical sound waves.

[0038] As noted above, one form of energy can be transformed into another. These energy forms may include, for example, mechanical, electrical, chemical, electromagnetic, thermal, and acoustic energy. Research has been conducted to exploretransforming other forms of energy, such as transforming light energy (e.g., high-energy photons) to mechanical energy. Transforming light energy into some form of mechanical energy requires efficient momentum transfer, and that is difficult to attain. An efficient system that can perform such a transformation is desired.

[0039] Based on their operating principle, micropumps can be divided into two groups: mechanical and nonmechanical. Although nonmechanical micropumps have no moving parts, they still require carefully fabricated microstructures and electrical contacts to generate thermal, electrical, magnetic, or acoustic stimulus to drive the fluids. While the performance of micropumps improved as the fabrication technique evolved, the principle and design of micropumps have remained almost the same over past decades. In various embodiments, the micropump has no moving parts or electrodes, thus requires no micro- or nanofabrication. In various embodiments, the size, number, location, and timing of the micropumps can be remotely controlled, reconfigured, and programmed in real-time.

[0040] Cell-based therapies using engineered cells have the potential to transform treatment of diseases like cancer. However, efficient delivery of macromolecules into cells remains challenging due to (i) cell viability and cytotoxicity and (ii) poor delivery efficiencies. The present disclosure describes a method for cytosolic delivery of exogenous macromolecules using a photo-acoustic streaming phenomenon.

[0041] With reference to FIGS. 1 and 2, an apparatus 100 for membrane permeabilizati on-based cell delivery is shown. The apparatus 100 generally includes a substrate 110 with photoacoustic launch pads 102 (e.g., gold ion implanted regions) and a laser source 300 (FIG. 3).[0042| The substrate 1 10 may include quartz or glass. Implanted substrates are made by exposing the surface of a substrate 110 (e.g., glass or quartz) to a stream of high- energy metal ions (e.g., gold ions) created by a particle accelerator. In aspects, the photoacoustic launch pads 102 (i.e., nano-particle embedded areas) may be created by gold (Au) ion implantation in about a 0.5-mm-thick quartz substrate 110 at about 60 keV to a dose of about 6 * 1016per cm square. Based on the chosen acceleration voltage, the Au ions may be implanted within about 50 nm below the surface. A relatively high dose may be used so that a sufficient Au nanoparticle concentration and corresponding optical absorption can be obtained. In aspects, the metal ions may be implanted through a track- etched membrane that has pores of the desired diameter (e.g., about 50 nm) and / or by adhering the track-etched membrane to the surface of an already implanted substrate.

[0043] The laser source 300 is configured to generate laser energy 302 (e.g., an ion beam). It is contemplated that the laser energy 302 may be any wavelength in the range of about 180nm to about 1mm. The laser source 300 may include a fiberoptic element (not shown) configured to convey the laser energy 302 to the substrate 110. The end surface of the fiber optic element may include flat, dimpled (concave), or convex shapes. For example, the convex shapes may be used for defocusing the laser energy 302. The concave shape may be used to focus the laser energy 302 on a small target area. The fiber optic element may range in diameter between approximately a fraction of a micron to over several millimeters. FIG. 2 shows a diagram of the substrate 1 10 with the photoacoustic launch pads 102 and a cell 200 (e.g., a mammalian cell) in situ.

[0044] The disclosed apparatus 100 overcomes the challenges of current photoporation methods by directly patterning nano-scale gold implanted areas onbiocompatible materials (e.g., glass or quartz) to create photoporation substrates that are safe, self-contained, and controllable. The disclosed ion beam-based method for creating nano-scale patterns of gold nano-particle embedded areas that, when illuminated by the laser, lead to the formation of vapor nanobubble (VNB) based photoporation sites.

[0045] In aspects, the nano-patterned areas may be fabricated by taking a nanopatterned (e.g., track-etched) membrane and adhering it to the surface of an implanted substrate, instead of patterning the implantation itself.

[0046] Referring to FIG. 3, laser energy 302 being applied to the photoacoustic launch pads 102 of the apparatus 100, is shown. The laser source 300 generates pulsed laser energy 302 which is applied to the photoacoustic launch pads 102. The membrane 202 of the cell 200 is surrounded by a fluid 204.

[0047] With reference to FIG. 4, a vapor nanobubble 400 (VNB) is created by absorption of the pulsed laser energy 302 by the photoacoustic launch pads 102 of the substrate 110 that are in contact with or near the target cell 200.

[0048] Referring to FIGS. 5 and 6, the VNB 400 collapses, and a pore 206 is created in the membrane 202 of the cell 200.

[0049] With reference to FIG. 7, a payload 702 is delivered into the cell 200 via the newly created pore 206. The payload 702 (e.g., small molecules and / or macromolecules) is in the fluid 204.

[0050] Referring to FIG. 8, a method 800 for intracellular delivery of a payload (e.g. small molecules or macromolecules such as proteins or DNA / mRNA) through a pore 206 created in the cytosolic membrane 202 of a cell 200 is shown. The method uses a nanopatterned implanted substrate 110 and pulsed laser energy 302 (FIG. 3). The cellmembrane pore 206 is created in the membrane 202 by one of two mechanisms: (i) VNB photoporation, or ii) the disruption of the cell membrane 202 by nano-jets of fluid streaming induced by an ultrasound wave propagating from the photoacoustic launch pads 102 into the fluid 204 after absorbing the energy from the pulsed laser energy 302.

[0051] Initially, the cell 200 is disposed in a fluid 204. The fluid 204 may be water, blood, plasma, body fluid, or any other fluid, depending on the medical or surgical application.

[0052] At step 802, the apparatus generates laser energy from a laser source 300 (FIG. 3). The laser source may generate laser energy in wavelengths in the range of about 180nm to about 1mm.

[0053] At step 804, VNBs 400 are created by the absorption of the pulsed laser energy 302 by the photoacoustic launch pads 102 of the substrate 110 that are in contact with or near the target cell 200 (FIG. 4). In a VNB 400, temperature gradients in the fluid can reach hundreds of degrees kelvin per nanometer.

[0054] At step 806, the apparatus increases the temperature of the photoacoustic launch pads 102 based on the absorption of the laser energy. At step 808, the apparatus creates a vapor nanobubble by causing fluid surrounding the cell to absorb heat and expand in response to the increased temperature of the photoacoustic launch pads 102. For example, the photoacoustic launch pads 102 absorb the laser energy and increase in temperature, which evaporates nearby water molecules, creating a VNB 400 (FIGS. 4 and 5).

[0055] At step 810, upon cessation of application of laser energy, the vapor nanobubble collapses, disrupting the membrane of the cell. At step 812, a pore 206 formsin the cell membrane 202 in response to the VNB 400, disrupting the membrane 202 of the cell 200 (FIG. 6).

[0056] At step 814, a payload 702 is delivered into the cell from the fluid surrounding the cell containing the payload 702. The payload 702 enters the cell via the pore 206. In aspects, the payload 702 may include small molecules or macromolecules, such as protein or nucleic acid.

[0057] For example, when the VNB collapses, this creates a pore 206 in the membrane of the cell (FIG. 6). This enables the surrounding fluid containing the payload to enter the cell (FIG. 7). After the membrane 202 recovers from the transient pore 206, the cell 200 has been transfected with the payload 702.

[0058] In aspects, the apparatus 100 disrupts the cell membrane 202 by nano-jets of fluid streaming induced by an ultrasound wave propagating from the photoacoustic launch pads 102 (i.e., the nano-patterned areas) into the fluid after the photoacoustic launch pads 102 absorb the energy from the pulsed laser energy 302. This mechanism may be described as a laser-driven photoacoustic microfluid pump (LDMP).

[0059] In aspects, the apparatus may consist of a microfluidic channel through which cells flow in a single fde. When they reach the treatment location, a laser pulse is directed to the wall of the microfluidic channel. The wall of the channel is patterned with photoacoustic launch pads (i.e., implanted nanopattems) that absorb the laser energy 302 and evaporate the nearby fluid 204 (e.g., water molecules) (FIG. 3) to create a vapor nanobubble 400 (FIG. 4). In aspects, one or more sides and / or the bottom of the channel may include the photoacoustic launch pads. When the vapor nanobubble 400 collapses, this creates a pore in the membrane 202 of the cell 200). This enables the surroundingfluid 204, containing the payload 702 (e.g., small molecules or macromolecules such as protein or nucleic acid) to enter the cell 200 (FIG. 7). After the membrane recovers from the transient pore, the cell 200 has been transfected with the payload 702. In another embodiment, a substrate is patterned with an array of nanometer spots. The cells are either deposited or flowed over the substrate. Illumination of the substrate by a pulsed laser creates the VNB that, in turn, creates pores in the surface of mammalian cells and enables supernatant cargo (e.g., macromolecules including protein, DNA, mRNA) to be introduced into the cells.[0060| The jets launched by this micropump from the substrate 110 (FIG.1) always flow normal to the substrate 110 surface regardless of the direction of laser beams. A simple conclusion from this observation is that the jets are not driven by momentum transfer from incident photons. Because the laser-induced heating and subsequent photothermal expansion of the Au-implanted quartz layer do not depend on the angle of incident lasers, the observation of jets perpendicular to the surface agrees with the mechanism of photoacoustic streaming and does not contradict previous observations. On the other hand, this substrate-jet relationship enables the micropump to pump fluids in the same direction without worrying about the direction of a laser beam. The direction of pumping may be changed with the direction of the substrate 110 (FIG. 1).[0061[ Laser-induced heating and photothermal expansion of the embedded Au particles induce stress inside the quartz plate and cause ultrasonic vibrations of the quartz plate. Such a local surface vibration and heating induce vibration of water in two directions: longitudinal vibration normal to the quartz surface and shear vibration parallel to the surface. However, the shear motion of the plate cannot be effectively propagatedinto the fluid because this motion decays exponentially in the fluid, and the penetration distance is typically less than 1pm at room temperature. Only the longitudinal vibrations of the quartz plate can be effectively propagated through the fluid. This vibration induces a longitudinal ultrasonic wave in the fluid, leading to a fluid jet normal to the plate surface.

[0062] An LDMP is made by ion implantation of gold atoms (or other metallic atoms) into a solid substrate such as quartz, glass, or other transparent materials. The substrate size can be small, for example, a sub-mm size. The Au may be implanted into a large, thin substrate.

[0063] Example 1. An apparatus for payload delivery to a cell, the apparatus comprising: a container for holding a fluid containing the payload and the cell to which the payload is to be delivered; a laser source configured to generate laser energy; and a substrate implanted with photoacoustic launch pads, the photoacoustic launch pads configured to absorb laser energy generated by the laser source.

[0064] Example 2. The apparatus of Example 1, wherein the photoacoustic launch pads are configured to increase in temperature based on the absorbed laser energy to create a vapor nanobubble in the fluid.

[0065] Example 3. The apparatus of Example 2, wherein the vapor nanobubble creates a pore in a cell membrane of the cell thereby enabling the payload contained in the fluid to enter the cell.

[0066] Example 4. The apparatus of Example 1, wherein the photoacoustic launchpads are configured to: induce a nano-jet of streaming fluid, propagating from the photoacoustic launch pads towards the cell, in response to the photoacoustic launch pads absorbing the laser energy; and cause a disruption of a cell membrane by the nano-jet of streaming fluid.(0067] Example 5. The apparatus of Example 1, wherein the substrate is at least one of quartz or glass.

[0068] Example 6. The apparatus of Example 1, wherein the photoacoustic launch pads comprise a nanopattem of metal ions on the substrate.

[0069] Example 7. The apparatus of Example 1, wherein the photoacoustic launch pads include a nanopattem of metal ions.

[0070] Example 8. The apparatus of Example 7, wherein the metal ions include gold ions.(0071 ] Example 9. The apparatus of Example 1, wherein the container is a microfluidic channel through which cells flow in single file.

[0072] Example 10. The apparatus of Example 9, wherein the substrate is part of a wall of the microfluidic channel.

[0073] Example 11. The apparatus of Example 1, wherein the laser source is configured to generate pulsed laser energy.

[0074] Example 12. A method for payload delivery to a cell, the method comprising: applying laser energy to a substrate to heat photoacoustic launch pads implanted within a substrate, the substrate proximate to a fluid, the fluid containing the cell, the photoacoustic launch pads configured to absorb the laser energy; and in response to the photoacoustic launch pads absorbing the applied laserenergy, creating a pore in a cell membrane of a cell contained in the fluid.

[0075] Example 13. The method of Example 12, further comprising: increasing a temperature of the photoacoustic launch pads in response to absorption of the laser energy; and creating a vapor nanobubble by causing fluid surrounding the cell to absorb heat and expand in response to the increased temperature of the photoacoustic launch pads.

[0076] Example 14. The method of Example 13, further comprising: causing the vapor nanobubble to collapse, disrupting the cell membrane.

[0077] Example 15. The method of Example 14, further comprising: forming a pore in the cell membrane in response to disrupting the membrane of the cell.

[0078] Example 16. The method of Example 15, further comprising: delivering a payload into the cell based on causing fluid surrounding the cell containing the payload to enter the cell via the pore.

[0079] Example 17. The method of Example 12, further comprising: inducing nano-jet of streaming fluid, propagating from the photoacoustic launch pads towards the cell, in response to the photoacoustic launch pads absorbing the energy from the laser energy; and

[0080] disrupting a cell membrane by the nano-jet of streaming fluid.

[0081] Example 18. A laser-driven photoacoustic microfluid pump comprising: a substrate implanted with photoacoustic launch pads, wherein the photoacoustic launch pads are configured to increase intemperature in response to exposure to laser energy, and

[0082] wherein the photoacoustic launch pads, when disposed proximate to a fluid, are further configured to induce a nano-jet of streaming fluid in response to the photoacoustic launch pads absorbing the energy from the laser energy.(0083] Example 19. The laser-driven photoacoustic microfluid pump of Example 18, wherein the photoacoustic launch pads are configured to receive laser energy of a wavelength in a range of about 180nm to about 1mm.

[0084] Example 20. The laser-driven photoacoustic microfluid pump of Example 18, wherein the photoacoustic launch pads include a nanopattern of gold ions, and wherein the substrate is at least one of quartz or glass.

[0085] The embodiments disclosed herein are examples of the disclosure and may be embodied in various forms. For instance, although certain embodiments herein are described as separate embodiments, each of the embodiments herein may be combined with one or more of the other embodiments herein. Specific structural and functional details disclosed herein are not to be interpreted as limiting, but as a basis for the claims and as a representative basis for teaching one skilled in the art to variously employ the present disclosure in virtually any appropriately detailed structure. Like reference numerals may refer to similar or identical elements throughout the description of the figures.

[0086] The phrases “in an embodiment,” “in embodiments,” “in various embodiments,” “in some embodiments,” or “in other embodiments” may each refer to one or more of the same or different embodiments in accordance with the present disclosure. A phrase in the form “A or B” means “(A), (B), or (A and B).” A phrase in the form “at least one of A, B, or C” means “(A); (B); (C); (A and B); (A and C); (B and C); or (A, B, and C) .”

[0087] It should be understood that the foregoing description is only illustrative of the present disclosure. Various alternatives and modifications can be devised by those skilled in the art without departing from the disclosure. Accordingly, the present disclosure is intended to embrace all such alternatives, modifications and variances. The embodiments described with reference to the attached drawing figures are presented only to demonstrate certain examples of the disclosure. Other elements, steps, methods, and techniques that are insubstantially different from those described above and / or in the appended claims are also intended to be within the scope of the disclosure.

Claims

CLAIMSWhat is claimed is:

1. An apparatus for payload delivery to a cell, the apparatus comprising: a container for holding a fluid containing the payload and the cell to which the payload is to be delivered; a laser source configured to generate laser energy; and a substrate implanted with photoacoustic launch pads, the photoacoustic launch pads configured to absorb laser energy generated by the laser source.

2. The apparatus of claim 1, wherein the photoacoustic launch pads are configured to increase in temperature based on the absorbed laser energy to create a vapor nanobubble in the fluid.

3. The apparatus of claim 2, wherein the vapor nanobubble creates a pore in a cell membrane of the cell thereby enabling the payload contained in the fluid to enter the cell.

4. The apparatus of claim 1, wherein the photoacoustic launch pads are configured to: induce a nano-jet of streaming fluid, propagating from the photoacoustic launch pads towards the cell, in response to the photoacoustic launch pads absorbing the laser energy; and cause a disruption of a cell membrane by the nano-jet of streaming fluid.

5. The apparatus of claim 1, wherein the substrate is at least one of quartz or glass.

6. The apparatus of claim 1 , wherein the photoacoustic launch pads comprise a nanopattem of metal ions on the substrate.

7. The apparatus of claim 1, wherein the photoacoustic launch pads include a nanopattern of metal ions.

8. The apparatus of claim 7, wherein the metal ions include gold ions.

9. The apparatus of claim 1, wherein the container is a microfluidic channel through which cells flow in single file.

10. The apparatus of claim 9, wherein the substrate is part of a wall of the microfluidic channel.

11. The apparatus of claim 1, wherein the laser source is configured to generate pulsed laser energy.

12. A method for payload delivery to a cell, using the apparatus of claim 1, the method comprising: applying laser energy to the substrate to heat photoacoustic launch pads implanted within the substrate, the substrate proximate to a fluid, the fluid containing the cell, the photoacoustic launch pads configured to absorb the laser energy; and in response to the photoacoustic launch pads absorbing the applied laser energy, creating a pore in a cell membrane of a cell contained in the fluid.

13. The method of claim 12, further comprising: increasing a temperature of the photoacoustic launch pads in response to absorption of the laser energy; and creating a vapor nanobubble by causing fluid surrounding the cell to absorb heat and expand in response to the increased temperature of the photoacoustic launch pads.

14. The method of claim 13, further comprising: causing the vapor nanobubble to collapse, disrupting the cell membrane.

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