CN113021874A - Single cell printing method based on annular laser spot induced transfer - Google Patents

Single cell printing method based on annular laser spot induced transfer Download PDF

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Publication number
CN113021874A
CN113021874A CN202110224495.6A CN202110224495A CN113021874A CN 113021874 A CN113021874 A CN 113021874A CN 202110224495 A CN202110224495 A CN 202110224495A CN 113021874 A CN113021874 A CN 113021874A
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annular
laser
cells
ring
method based
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章宇健
林士楠
邓宇
郭钟宁
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Guangdong University of Technology
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Guangdong University of Technology
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
    • B29C—SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING
    • B29C64/00—Additive manufacturing, i.e. manufacturing of three-dimensional [3D] objects by additive deposition, additive agglomeration or additive layering, e.g. by 3D printing, stereolithography or selective laser sintering
    • B29C64/10—Processes of additive manufacturing
    • B29C64/106—Processes of additive manufacturing using only liquids or viscous materials, e.g. depositing a continuous bead of viscous material
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
    • B29C—SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING
    • B29C64/00—Additive manufacturing, i.e. manufacturing of three-dimensional [3D] objects by additive deposition, additive agglomeration or additive layering, e.g. by 3D printing, stereolithography or selective laser sintering
    • B29C64/20—Apparatus for additive manufacturing; Details thereof or accessories therefor
    • B29C64/264—Arrangements for irradiation
    • B29C64/268—Arrangements for irradiation using laser beams; using electron beams [EB]
    • B29C64/273—Arrangements for irradiation using laser beams; using electron beams [EB] pulsed; frequency modulated
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B33—ADDITIVE MANUFACTURING TECHNOLOGY
    • B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
    • B33Y10/00—Processes of additive manufacturing
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus

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  • Chemical & Material Sciences (AREA)
  • Materials Engineering (AREA)
  • Health & Medical Sciences (AREA)
  • Optics & Photonics (AREA)
  • Manufacturing & Machinery (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Mechanical Engineering (AREA)
  • Wood Science & Technology (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
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  • Biotechnology (AREA)
  • Sustainable Development (AREA)
  • Microbiology (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
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Abstract

The invention discloses a unicell printing method based on annular laser spot induced transfer, which comprises S1, focusing an annular spot on a sacrificial layer; s2, forming a plasma annular cavitation bubble on the sacrificial layer; s3, expanding the annular cavitation bubbles and pushing away cell solution outside the ring, wherein the target transfer cells in the center of the ring are pushed downwards; s4, completely pushing out the target transfer cells by the annular cavitation bubble collapse shock wave generated in the vertical direction of the center position of the ring; s5, the target metastatic cells and the entrained solution are printed. The invention utilizes the central convergent vertical shock wave generated after the annular cavitation bubble is collapsed to push the printing of the biological cell solution, so that target transfer cells are not directly irradiated by laser to generate ablation; and the fluid mechanics interaction among cells is blocked by utilizing the pushing effect of the expansion of the annular cavitation bubbles on the non-target transferred biological solution outside the ring, so that the annular cavitation bubbles induced by the annular light spots effectively block substances inside and outside the ring, and the quantity of the target transferred substances is accurate and controllable.

Description

Single cell printing method based on annular laser spot induced transfer
Technical Field
The invention relates to a cell printing method, in particular to a unicell printing method based on annular laser spot induced transfer.
Background
The three-dimensional biological printing technology is a flexible, automatic and on-demand complex biological structure manufacturing platform, and is a novel human organ and tissue design and engineering technology. The existing 3D bio-printing technologies can be mainly classified into the following three categories: inkjet printing, squeeze printing, and laser induced bioprinting.
Inkjet printing is the deposition of cell droplets or bio-ink on a receiving layer in droplet-based bio-printing methods, including inkjet bio-printing, micro-valve based bio-printing, and acoustic based bio-printing. Of the different types of inkjet printing techniques, only thermal and piezoelectric drop-on-demand printing techniques are commonly used for bioprinting. In thermal bioprinting, the application of a voltage pulse heats the bio-ink, resulting in localized heating; after the vapor bubble is formed, the bubble rapidly expands and disintegrates, creating pressure within the bio-ink reservoir, helping the bio-ink to overcome the surface tension of the nozzle tip, thereby ejecting a water droplet. In the piezoelectric bioprinting technique, a voltage pulse is applied to a piezoelectric crystal, which expands, thereby generating a pressure wave in a biological solution pool to help the bio-ink overcome the surface tension of the nozzle tip and eject a droplet. The advantages of ink jet printers are fast printing speed, low cost and wide usability. However, the risk of exposure of cells and materials to thermal and mechanical stress, low droplet directionality, non-uniform droplet size, frequently clogged nozzles, pose considerable disadvantages for 3D bioprinting using these printers.
Extrusion bioprinting bio-ink containing cells is extruded out of the nozzle by air pressure (pneumatic) or mechanical systems (piston or screw). This is the simplest of all bioprinting methods. The method has the advantages of strong load capacity, high printing speed, strong expandability and wide printing capability, and the range of the biological ink (hydrogel, polymer, microcarrier, acellular ECM and cell aggregation) is one of the methods. However, these high pressures adversely affect cell viability due to nozzle shear forces, particularly for high viscosity materials, resulting in cell viability typically between 40% and 95%.
Laser induced bioprinting (LIFT) is a non-contact, jet-less printing that uses laser energy to eject droplets of cellular bio-ink onto a receiving substrate. Laser-based bioprinting techniques come in different types, with the same basic principles, but with slightly different experimental settings. The size of the bio-ink voxels or droplets formulated by this technique depends on several process parameters. Including laser pulse energy, focused laser spot size, laser pulse duration, distance between donor and collector slides, thickness of laser energy absorbing layer and cell bio-ink layer, etc. LIFT has many advantages. First, it is a nozzle-less approach, eliminating the problems of nozzle clogging and contamination risks. Secondly, high resolution can be achieved, enabling printing of high density cell suspensions and bio-inks (1-300mPa/s) with a wide range of viscosities. LIFT also has many limitations, and the fluidic effect driven by the circular cavitation bubbles induced by the existing LIFT technology may cause non-target metastatic cells to be printed together due to the complex hydrodynamic properties of the cell suspension. The main disadvantage of the existing LIFT technology is the side effect of laser irradiation on cell activity.
Therefore, it is necessary to design a printing method that solves the problems of the conventional laser induced bio-printing technique that the conveying precision is not high, and the printing efficiency and mechanical stress caused by the nozzle clogging are solved without the nozzle.
Disclosure of Invention
The invention aims to solve the problems and provides a unicell printing method based on annular laser spot induced transfer. The method combines the annular cavitation bubbles generated by annular light spot induction with a laser induction technology, avoids the direct irradiation process of laser on target transfer cells by utilizing the special structure of the annular light spot, and simultaneously, the special structure of the annular cavitation bubbles can play a role in blocking the transfer of non-target transfer cells, thereby solving the problems of low transfer precision of the traditional laser-induced biological printing technology, and low printing efficiency and mechanical stress caused by nozzle blockage without a nozzle.
The purpose of the invention can be achieved by adopting the following technical scheme:
a unicell printing method based on annular laser spot induced transfer comprises the following steps:
step 1, after pulse laser beams which are uniformly distributed are incident to an annular light spot forming system element, emergent light is focused to form an annular light spot on a sacrificial layer through a transparent constraint layer;
step 2, the sacrificial layer generates ablation under the action of light and heat of laser and forms a plasma annular cavitation bubble;
step 3, rapidly expanding the annular cavitation bubbles and pushing away cell solution outside the rings, and meanwhile, pushing down target transfer cells in the centers of the rings under the expansion of the cavitation bubbles;
step 4, collapsing the annular cavitation bubbles, and completely pushing out target transfer cells by collapse shock waves generated in the direction vertical to the center position of the ring;
and 5, under the action of gravity and self momentum, printing the target transfer cells and the carried solution on a receiving plate.
Further, the specific content in step 1 includes: through changing the inner and outer diameter of the annular light spot, the laser energy value and the distance between the transparent constraint layer and the receiving plate, the biological cells with different diameters are accurately printed.
Further, the material of the sacrificial layer in step 2 is a material that can be photolyzed or pyrolyzed by laser and generates gas.
Preferably, the sacrificial layer is triazene or metallic titanium.
The implementation of the invention has the following beneficial effects:
1. the invention combines the annular cavitation bubbles generated by the induction of the annular light spots with the laser-induced forward transfer technology, and uses the central convergent vertical shock wave generated after the annular cavitation bubbles are collapsed to push the printing of the biological cell solution, so that target transfer cells are not directly irradiated by laser to generate ablation; and the fluid mechanics interaction among cells is blocked by utilizing the pushing effect of the annular cavitation bubble expansion stage on the non-target transferred biological solution outside the ring, so that the annular cavitation bubble induced by the annular light spot effectively blocks substances inside and outside the ring, the quantity of the target transferred substances is accurate and controllable, and the problem of low transmission precision of the traditional laser-induced biological printing technology is solved.
2. The invention does not need a nozzle, has simpler and more convenient operation and high efficiency, does not have the problem of nozzle blockage, has high deposition efficiency, does not have the mechanical stress of the nozzle acting on cells, and has wide viscosity range of printable cell solution.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a schematic flow chart of a single cell printing method based on annular laser spot induced transfer according to the present invention;
FIG. 2 is a schematic diagram of a single cell printing method based on annular laser spot induced transfer shooting annular laser spot induced annular cavitation forward transfer process by high-speed photography;
FIG. 3 is a schematic diagram of a single cell printing method based on annular laser spot induced transfer, which shoots an annular laser spot through high-speed photography to induce the transfer process of sodium alginate cell solution.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example (b):
referring to fig. 1 to 3, the present embodiment relates to a single cell printing method based on annular laser spot induced transfer, including the following steps:
step 1, after pulse laser beams 1 which are uniformly distributed are incident to an annular light spot forming system element 2, emergent light is focused to form an annular light spot 3 on a sacrificial layer 5 through a transparent constraint layer 4; through changing the inner and outer diameter of the annular light spot 3, the laser energy value and the distance between the transparent constraint layer 4 and the receiving plate 9, the single biological cells with different diameters are accurately printed.
Step 2, the sacrificial layer 5 generates ablation under the action of light and heat of laser and forms a plasma annular cavitation bubble 7; the material of the sacrificial layer 5 is a material which can be photolyzed or pyrolyzed by laser and generates gas, for example, triazene or metallic titanium can be used as the sacrificial layer 5.
Step 3, rapidly expanding the annular cavitation bubbles 7 and pushing away cell solution outside the rings, and meanwhile, pushing down target transfer cells in the centers of the rings under the expansion of the cavitation bubbles;
step 4, the annular cavitation bubbles 7 are collapsed, and collapse shock waves generated in the direction vertical to the center of the ring push out the target transfer cells 8 completely;
step 5, under the action of gravity and self momentum, the target transfer cell 8 and the carried solution are printed on a receiving plate.
The method combines the annular cavitation bubbles 7 generated by the induction of the annular light spots 3 with a laser-induced forward transfer technology, and uses the central convergent vertical shock wave generated after the annular cavitation bubbles 7 are collapsed to push the printing of the biological cell solution, so that the target transfer cells 8 are not directly irradiated by laser to generate ablation; and the fluid mechanics interaction between cells is blocked by the pushing effect of the annular cavitation bubbles 7 on the non-target transferred biological solution outside the ring in the expansion stage, so that the annular cavitation bubbles 7 induced by the annular light spots 3 effectively block substances inside and outside the ring, the amount of the target transferred substances is accurate and controllable, and the problem of low transmission precision of the traditional laser-induced biological printing technology is solved.
The method has the advantages of no need of a nozzle, simpler and more convenient operation and high efficiency, and solves the problems of low printing efficiency and mechanical stress caused by nozzle blockage.
While the invention has been described in connection with what is presently considered to be the most practical and preferred embodiment, it is to be understood that the invention is not to be limited to the disclosed embodiment, but on the contrary, is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims.

Claims (4)

1. A unicell printing method based on annular laser spot induced transfer is characterized by comprising the following steps:
step 1, after pulse laser beams which are uniformly distributed are incident to an annular light spot forming system element, emergent light is focused to form an annular light spot on a sacrificial layer through a transparent constraint layer;
step 2, the sacrificial layer generates ablation under the action of light and heat of laser and forms a plasma annular cavitation bubble;
step 3, rapidly expanding the annular cavitation bubbles and pushing away cell solution outside the rings, and meanwhile, pushing down target transfer cells in the centers of the rings under the expansion of the cavitation bubbles;
step 4, collapsing the annular cavitation bubbles, and completely pushing out target transfer cells by collapse shock waves generated in the direction vertical to the center position of the ring;
and 5, under the action of gravity and self momentum, printing the target transfer cells and the carried solution on a receiving plate.
2. The single-cell printing method based on annular laser spot induced transfer as claimed in claim 1, wherein the specific content in step 1 includes: through changing the inner and outer diameter of the annular light spot, the laser energy value and the distance between the transparent constraint layer and the receiving plate, the biological cells with different diameters are accurately printed.
3. The single-cell printing method based on ring-shaped laser spot induced transfer as claimed in claim 1, wherein the material of the sacrificial layer in step 2 is a material that can be photolyzed or pyrolyzed by laser and generate gas.
4. The single-cell printing method based on annular laser spot induced transfer as claimed in claim 1 or 3, wherein the sacrificial layer is triazene or metallic titanium.
CN202110224495.6A 2021-03-01 2021-03-01 Single cell printing method based on annular laser spot induced transfer Pending CN113021874A (en)

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114178547A (en) * 2021-07-22 2022-03-15 广东工业大学 Laser-induced transfer printing method for micro electronic elements based on non-Newtonian fluid characteristics
CN114551324A (en) * 2022-04-25 2022-05-27 浙江清华柔性电子技术研究院 Transfer method of micro device
WO2024084031A1 (en) 2022-10-20 2024-04-25 Poietis Printing objects from a well
WO2024084030A1 (en) 2022-10-20 2024-04-25 Poietis Object manipulation and transfer by propulsion

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000034757A1 (en) * 1998-12-10 2000-06-15 The Government Of The Unites States Of America Represented By The Secretary, Department Of Health And Human Services Designs for non-contact laser capture microdissection
CN110484425A (en) * 2019-08-30 2019-11-22 东南大学 A kind of unicellular acquisition device

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000034757A1 (en) * 1998-12-10 2000-06-15 The Government Of The Unites States Of America Represented By The Secretary, Department Of Health And Human Services Designs for non-contact laser capture microdissection
CN110484425A (en) * 2019-08-30 2019-11-22 东南大学 A kind of unicellular acquisition device

Non-Patent Citations (1)

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Title
丁世鹏: "激光捕获技术及环形光束的聚焦特性研究", 《中国优秀博硕士学位论文全文数据库(硕士) 信息科技辑》 *

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114178547A (en) * 2021-07-22 2022-03-15 广东工业大学 Laser-induced transfer printing method for micro electronic elements based on non-Newtonian fluid characteristics
CN114551324A (en) * 2022-04-25 2022-05-27 浙江清华柔性电子技术研究院 Transfer method of micro device
WO2024084031A1 (en) 2022-10-20 2024-04-25 Poietis Printing objects from a well
WO2024084030A1 (en) 2022-10-20 2024-04-25 Poietis Object manipulation and transfer by propulsion
FR3141091A1 (en) * 2022-10-20 2024-04-26 Poietis PRINTING OBJECTS FROM A WELL
FR3141092A1 (en) * 2022-10-20 2024-04-26 Poietis HANDLING AND TRANSFER OF OBJECTS BY PROPULSION

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Application publication date: 20210625