CN107418998B - Microparticle manipulation method, bioprinting method and biological construct - Google Patents
Microparticle manipulation method, bioprinting method and biological construct Download PDFInfo
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- CN107418998B CN107418998B CN201610822794.9A CN201610822794A CN107418998B CN 107418998 B CN107418998 B CN 107418998B CN 201610822794 A CN201610822794 A CN 201610822794A CN 107418998 B CN107418998 B CN 107418998B
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Abstract
The invention relates to the technical field of particle manipulation, in particular to a particle manipulation method. The method comprises forming a liquid film on the lower end of a suction member; and step b: the fine particles are sucked by the sucking member, and the fine particles are held at the lower end of the sucking member and brought into contact with the liquid film. Compared with the existing microparticle manipulation method, the microparticle manipulation method provided by the invention can effectively reduce the damage to the microparticles by forming the liquid film on the lower end part of the suction part in advance and keeping the microparticles at the position where the lower end part of the suction part is contacted with the liquid film, thereby being beneficial to keeping the preset shape and/or good biological activity of the microparticles.
Description
Technical Field
The invention relates to the technical field of particle manipulation, in particular to a particle manipulation method, a biological printing method and a biological construct.
Background
In the prior art, pipettes such as a pipette gun, a dropper and a syringe are usually adopted to control particles, when the pipette is used, the pipette gun with a proper volume, the dropper with a proper needle caliber or the syringe and the like are selected according to the size of the particles, the pipettes are manually operated to suck the particles into a tube cavity of the corresponding pipettes by means of negative pressure generated when the pipettes suck the liquid, so that the particles are captured, and after the pipettes are displaced, the particles are placed at a set placement position by means of positive pressure generated when the pipettes discharge the liquid, so that the particles are placed.
However, since the existing microparticle manipulation method needs to suck microparticles into the lumen of the pipette, it is easy to damage the microparticles, for example, the microparticles cannot maintain the preset shape, and for example, for bioactive microparticles, the inner wall of the lumen is damaged due to friction, causing cell damage or death, and affecting the bioactivity of the microparticles, so as to affect the microparticles to achieve the corresponding biological function, and finally, affect the biological function of the biological construct formed thereby. Moreover, the existing particle control method completely depends on the negative pressure of a liquid shifter to realize the suction of the particles, and a larger suction force is required to be applied, so that on one hand, the requirement on the suction technology is improved, the suction difficulty is increased, and the control success rate is reduced, and on the other hand, the larger suction force can further aggravate the damage to the particles; and, rely on the negative pressure to realize the absorption to the particle alone, absorb the controllability relatively poor, often appear a certain a plurality of particles of absorption, the condition that can't absorb the particle another time, it is difficult to guarantee the uniformity of absorbing at every turn, this also causes unnecessary trouble to control steps such as subsequent aversion and/or placing.
It should be noted that what is described in this section is for convenience of understanding only and should not be taken as an acknowledgement or suggestion that such matter is prior art.
Disclosure of Invention
The invention aims to solve the technical problems that: the existing particle control method is easy to cause damage to the particles.
In order to solve the above technical problem, a first aspect of the present invention provides a method for manipulating microparticles, which sequentially comprises the following steps:
a, forming a liquid film at the lower end part of the suction component; and
step b: the fine particles are sucked by a suction member having a liquid film formed on the lower end portion thereof, and are held on the lower end portion of the suction member and brought into contact with the liquid film.
Alternatively, in step a, the lower end portion of the sucking member is formed into a liquid film by sucking the liquid in advance with the sucking member.
Optionally, in step a, the liquid previously sucked by the sucking means is a liquid in which the particles are immersed.
Alternatively, in step b, after the fine particles are sucked to the lower end portion of the suction member by the suction member, the suction operation is stopped, and the fine particles are held at the lower end portion of the suction member by the negative pressure and the tension of the liquid film.
Alternatively, in step b, the microparticles are sucked by one or at least two sucking parts having lower end portions forming a liquid film, each sucking part sucking only a single microparticle.
Optionally, the particle manipulation method further comprises a step c, which is disposed after the step b: the suction member is moved to a position above the set placement position of the fine particles, and the fine particles are driven by the suction member to be separated from the lower end portion of the suction member and placed at the set placement position.
Alternatively, in step c, the suction member is moved downward to bring the fine particles into direct contact with the set placement position, and the fine particles are separated from the lower end portion of the suction member by the adhesion force with the set placement position and the gravity thereof and placed at the set placement position.
Alternatively, in step c, the fine particles are ejected by the suction member, and the fine particles are separated from the lower end portion of the suction member by the positive pressure and the self gravity and are placed at the set placement position.
Alternatively, when the fine particles are placed at the set placement position by the suction means, a part of the liquid sucked in advance remains in the suction means.
Optionally, the particle manipulation method further comprises a step d, which is disposed after the step c: the remaining liquid preliminarily sucked in the sucking means is discharged.
Optionally, the particles have a particle size of 0.5 to 3 mm.
Alternatively, the microparticles are gel-state microparticles capable of maintaining their own structural stability in the natural state.
Optionally, the particulate is a bio-brick.
The second aspect of the invention also provides a bioprinting method. The bioprinting method utilizes the particle manipulation method of the present invention to manipulate biological tiles as particles.
The third aspect of the invention also provides a biological construct. The biological construct is prepared using the bioprinting method of the present invention.
Compared with the existing microparticle manipulation method, the microparticle manipulation method provided by the invention can effectively reduce the damage to the microparticles and help the microparticles to maintain the preset form and/or good biological activity by forming the liquid film at the lower end part of the suction part in advance and keeping the microparticles at the position where the lower end part of the suction part is contacted with the liquid film.
Other features of the present invention and advantages thereof will become apparent from the following detailed description of exemplary embodiments thereof, which proceeds with reference to the accompanying drawings.
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, and 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 these drawings without creative efforts.
Fig. 1 shows an embodiment of the method for operating a bioprinting unit according to the present invention, wherein fig. 1a and 1b show step a, fig. 1c shows step b, fig. 1d and 1e show step c, and fig. 1f shows step d.
In the figure:
10. a liquid transferring gun; 20. a biological brick; 30. a storage unit; 40. and printing the substrate.
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. The following description of at least one exemplary embodiment is merely illustrative in nature and is in no way intended to limit the invention, its application, or uses. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without any inventive step, are within the scope of the present invention.
Techniques, methods, and apparatus known to those of ordinary skill in the relevant art may not be discussed in detail but are intended to be part of the specification where appropriate.
In the description of the present invention, it is to be understood that the orientation or positional relationship indicated by the orientation words such as "front, rear, upper, lower, left, right", "lateral, vertical, horizontal" and "top, bottom", etc. are usually based on the orientation or positional relationship shown in the drawings, and are only for convenience of description and simplicity of description, and in the case of not making a reverse description, these orientation words do not indicate and imply that the device or element being referred to must have a specific orientation or be constructed and operated in a specific orientation, and therefore, should not be considered as limiting the scope of the present invention; the terms "inner and outer" refer to the inner and outer relative to the profile of the respective component itself.
In the description of the present invention, it should be understood that the terms "first", "second", etc. are used to define the components, and are used only for the convenience of distinguishing the corresponding components, and if not otherwise stated, the terms have no special meaning, and thus, should not be construed as limiting the scope of the present invention.
As used herein, the term "biological construct" refers to an artificially constructed, two-or three-dimensional structure containing cells. In certain preferred embodiments, the biological construct is a three-dimensional construct, a tissue precursor, an artificial tissue or an artificial organ.
As used herein, the term "bio-brick" is a biologically active modular unit for the construction of biological organisms, which when present alone, maintains its spatial configuration, and which can be used in a variety of fields, such as bioprinting (e.g., 3D bioprinting), tissue engineering, and regenerative medicine. Wherein, in the biological printing field of 3D, biological brick can regard as the basic unit of biological printing of 3D. The preferred structure and composition of the bio-brick is: a nuclear layer comprising cells, wherein the cells are capable of growing, proliferating, differentiating or migrating, the nuclear layer being made of a biodegradable material and providing a substance required for the vital activity of the cells; and a shell layer encapsulating the core layer, the shell layer being located outside the core layer, being made of a biodegradable material, and providing mechanical protection for the core layer and the cells inside. The bio-brick of this preferred structure behaves as a bioactive microsphere.
As used herein, the term "gel-state particles" is a generic term for a class of particles having similar properties to biological bricks, which are capable of maintaining their own structural stability in the natural state.
Figure 1 illustrates one embodiment of the particle manipulation method of the present invention. Referring to fig. 1, the method for manipulating particles provided by the present invention sequentially comprises the following steps:
a, forming a liquid film at the lower end part of the suction component;
step b: the fine particles are sucked by the suction member having a liquid film formed on the lower end portion thereof, and are held on the lower end portion of the suction member and brought into contact with the liquid film.
According to the particle control method, when the suction component is used for capturing and obtaining the particles, the particles are not sucked into the tube cavity, but are kept at the lower end part of the suction component, and the abrasion of the inner wall of the tube cavity to the particles and the change of the particle form can be avoided, so that the damage of the tube cavity of the suction component to the particles can be effectively prevented; in addition, according to the present invention, a liquid film for contacting the fine particles is formed in advance on the lower end portion of the suction member before the fine particles are sucked to the lower end portion of the suction member, and the fine particles can be further isolated from the lower end portion of the suction member by the liquid film to indirectly contact the fine particles with the lower end portion of the suction member, so that the fine particles can be prevented from being mechanically damaged due to direct rigid contact with the lower end portion of the suction member, and damage to the fine particles during manipulation can be further reduced. In addition, the invention does not suck the particles into the tube cavity any more, but keeps the particles at the lower end part of the sucking part, thereby improving the controllability of sucking, ensuring the consistency of sucking operation every time more easily and facilitating the implementation of subsequent displacement and/or placement.
Compared with the existing microparticle manipulation method, the microparticle manipulation method provided by the invention has the advantages that the liquid film is formed at the lower end part of the suction part in advance, and the microparticles are kept at the position where the lower end part of the suction part is in contact with the liquid film, so that the damage to the microparticles can be effectively reduced, and the microparticles are helped to keep the preset shape and/or good biological activity.
In addition, the present invention has advantages that the liquid film is formed at the lower end part of the absorbing component in advance: (1) the preformed liquid film can play a role in attracting the particles to be sucked, so that the particles are not only acted by the negative pressure of the sucking part but also acted by the surface tension of the liquid film in the sucking process, on one hand, the negative pressure required to be applied by the sucking part can be reduced, the sucking technical requirement can be reduced, the successful sucking probability can be increased, the damage of the sucking force to the particles can be reduced, on the other hand, the particles can be more firmly kept at the lower end part of the sucking part, the displacement of the particles is facilitated, and the stability and the reliability of the particle control process are improved; (2) if the liquid film is not formed in advance, the particle size must be larger than the lumen diameter of the suction part to achieve the purpose of keeping the particles at the lower end part of the suction part, which results in that only the particles with larger size can be kept at the lower end part of the suction part, and the particles with smaller size are difficult to be controlled based on the non-inhalation type particle obtaining mode, while the invention forms the liquid film at the lower end part of the suction part in advance, which can effectively overcome the requirement of the non-inhalation type particle obtaining mode on the particle size, expand the application range of the non-inhalation type particle obtaining mode, and enable more particles to be controlled on the premise of smaller damage, for example, the control of the particles with the particle size within the range of 0.5-3mm can be realized.
The formation of the liquid film in step a of the present invention may be performed in various ways, for example, by painting. Wherein preferably, the liquid film may be formed at a lower end portion of the sucking part by sucking the liquid in advance with the sucking part. The liquid film forming device has the advantages that the liquid film is more convenient to form, the liquid sucked into the sucking component can also wet the inner wall of the tube cavity of the sucking component, the liquid film can be formed at the lower end part of the sucking component and in the tube cavity, and the damage to particles can be further reduced.
Further, the microparticle manipulation method of the present invention may further include a step c provided after the step b: the suction member is moved to a position above the set placement position of the fine particles, and the fine particles are driven by the suction member to be separated from the lower end portion of the suction member and placed at the set placement position. Based on this, the particle manipulation method of the invention not only can realize the capture and acquisition of the particles, but also can realize the displacement and placement of the particles.
Wherein the placing of the particles can be achieved by direct placing. That is, in step c, the suction member may be moved downward to bring the fine particles into direct contact with the set placement position, so that the fine particles are separated from the lower end portion of the suction member by the adhesion force with the set placement position and the gravity thereof and placed at the set placement position. The particles are placed based on the direct placing mode, and only the sum of the adhesive force between the particles and the set placing position and the gravity of the particles is larger than the sum of the tension between the particles and the liquid film and the sum of the negative pressure.
In addition, the invention can also realize the placement of the particles by a spraying mode. That is, in step c, the fine particles are ejected by the suction member so that the fine particles are separated from the lower end portion of the suction member by the positive pressure and the self gravity and are placed at the set placement position. This mode of placing, the positive pressure when utilizing the suction means tapping places the particle in setting for and places the position, can effectively improve and place efficiency.
When the suction component places the particles at the set placing position, a part of the liquid sucked in advance still remains in the suction component, namely the suction component does not discharge the liquid sucked in advance in the suction component. The advantage of this operation is that the liquid at the set placement position can be reduced as much as possible, so that the particles themselves are mainly at the set placement position, and the influence of the liquid sucked in advance on the function of the particles at the set placement position is reduced. Based on this, further, the particle manipulation method of the present invention may further include a step d disposed after the step c: the remaining liquid preliminarily sucked in the sucking means is discharged. Through setting up step d, can be convenient for absorb the part and continue to realize controlling or other usefulness in addition another particle, raise the efficiency.
The method for manipulating the microparticles of the present invention can be applied to various microparticles, particularly to the body-shaped objects with obvious curved surfaces, such as hemispheres, ellipsoids or spheres, and particularly to the microparticles with soft and non-hard (such as non-solid and non-hard surfaces), wherein more preferably, the method for manipulating the microparticles of the present invention can be applied to the gel-state microparticles with bioactivity, such as biological bricks, and capable of maintaining the structural stability of the microparticles under the natural state.
The particle manipulation method of the present invention is further described below in conjunction with the embodiment shown in fig. 1. In this embodiment, the particle manipulation method of the present invention is applied to a bio-printing process, and the manipulation of the particles includes sucking, displacing, and placing the bio-brick 20, that is, the particles of this embodiment are the bio-brick 20; the biological bricks 20 are stored in the storage unit 30 in advance (for example, in each hole of the pore plate), and the biological bricks 20 are immersed in the liquid in advance in the storage unit 30, so that the biological bricks 20 can maintain good activity in the liquid environment; the suction part is a pipette gun 10, the pipette gun 10 is a controllable pressure pipette gun so as to control suction and discharge pressure, the pipette gun 10 can be but is not limited to have 5 gears, and each gear corresponds to different pressure and different liquid volume; the set placement position is located on the printing substrate 40, which in this embodiment may also be referred to as a set printing position, and the printing substrate 40 may be a three-dimensional structure (e.g., a cylinder such as a gyrometer) or a planar structure, which may be selected according to the desired biological construct to be formed. Specifically, in this embodiment, the bio-brick 20 with a particle size of 2mm is used as the operation target, and the liquid-transferring gun 10 with a bore diameter of 0.4mm, 0.5mm, or 1mm may be used as the liquid-transferring gun 10 suitable for this embodiment, but in other embodiments, the particle size of the bio-brick 20 and the bore diameter of the liquid-transferring gun 10 may also be of other dimensions as long as they are suitable for each other.
In this embodiment, the microparticle manipulation method comprises the following steps in sequence:
step a: as shown in fig. 1a, first, the pipette 10 is set to the pipette mode, and the shift position is set to 2 (for example, 50 μ l), so as to complete the setting of the pipette 10; then, as shown in fig. 1b, the pipette 10 is moved to a position above (preferably directly above) the bio-brick 20 stored in the storage unit 30, the position of the pipette 10 is adjusted, then, a pipetting process is started to start pipetting, the pipette 10 is used to aspirate the liquid immersing the bio-brick 20, the liquid level in the pipette 10 is raised to 1-2 steps (for example, 40 μ l), and the liquid aspirated in advance forms a liquid film at the lower end of the pipette 10;
step b: as shown in fig. 1c, the pipetting gun 10 continues to aspirate the bio-brick 20 to the lower end of the pipetting gun 10 under the action of the negative pressure of the pipetting gun 10 and the surface tension of the liquid film, and the bio-brick is contacted with the liquid film, at which time the aspiration operation is stopped, and after the aspiration operation is stopped, although the bio-brick 20 slightly moves downward under the action of the inertial flow of the liquid, the bio-brick 20 can still be held at the lower end of the pipetting gun 10 under the action of the negative pressure and the liquid film tension, and at which time the liquid level inside the pipetting gun 10 rises to 2 steps, and the capture and acquisition (also referred to as grasping) of the bio-brick 20 is completed;
c, performing a step; as shown in fig. 1d, the pipette gun 10 holding the bio-brick 20 at the lower end is moved to the printing substrate 40, and the pipette gun 10 is positioned above (preferably directly above) the set printing position corresponding to the bio-brick 20, thereby completing the displacement of the bio-brick 20; then, as shown in fig. 1e, the pipette gun 10 is adjusted to the liquid discharging mode, the liquid discharging position is set between 1-2 (for example, 10 μ l out of 50 μ l), the liquid discharging is started, the biological brick 20 is separated from the lower end of the pipette gun 10 under the action of the positive pressure of the pipette gun 10 and the self gravity of the biological brick 20, and the biological brick falls to the set printing position, and the placement of the biological brick 20, that is, the printing of the biological brick 20 is completed;
step d: as shown in FIG. 1f, the pipette 10, with the previously aspirated liquid remaining inside, is removed from the printing substrate 40 and the pipette 10 is moved to a suitable discharge position, such as a dedicated waste reservoir, to discharge the remaining liquid in the pipette 10 (e.g., 40. mu.l of the remaining 50. mu.l) to complete the discharge of the remaining liquid.
In this embodiment, the liquid previously sucked into the pipette gun 10 can wet the inner wall of the muzzle and the muzzle tip of the pipette gun 10, so that a liquid film is formed at the muzzle of the pipette gun 10 (i.e. the lower end of the suction unit), and the liquid film can prevent the muzzle from directly contacting with the bio-brick 20 when the pipette gun 10 continues to suck the bio-brick 20, thereby preventing the form and biological activity of the bio-brick 20 from being damaged and destroyed due to the direct rigid contact with the muzzle.
Moreover, because the liquid film can separate the muzzle from the biological bricks 20, even the biological bricks 20 with the particle size smaller than the diameter of the muzzle can be kept at the muzzle under the action of the liquid film and the negative pressure, therefore, the requirement of a non-inhalation particle acquisition mode on the particle size of the biological brick 20 can be effectively overcome by arranging the liquid film, the application range of the non-inhalation particle acquisition mode is expanded, so that more specifications and types of bio-bricks 20 can be manipulated with less damage, and because of the smaller particle size of the bio-bricks 20, the number of cells contained inside is relatively small and the demand for nutrient supply is relatively low, so that the non-inhalational access to the bio-brick 20 with smaller particle size is achieved by using a liquid film, which is also beneficial to better maintain the biological activity of the bio-brick 20 during the printing process and is helpful to finally obtain a biological construct with better biological activity.
Moreover, the liquid film formed in advance at the muzzle can attract the biological brick 20 sucked later, so that the biological brick 20 is not only subjected to negative pressure but also subjected to the surface tension of the liquid film, on one hand, the negative pressure required to be applied by the liquid-transferring gun 10 can be reduced, the technical requirement on suction can be reduced, the successful suction probability is increased, and the damage of suction force to the biological brick 20 can be reduced; on the other hand, the biological bricks 20 can be firmly kept at the muzzle, so that the firmness in the grabbing process is improved, the biological bricks 20 can be firmly kept at the muzzle in the shifting process and are not easy to fall off, and the stability and the reliability in the shifting process are improved; on the other hand, the liquid film can make the bio-brick 20 always in the liquid environment from the process of being sucked out from the storage unit 30 to being placed at the set printing position, and the liquid environment is an important factor for keeping the bioactivity of the bio-brick 20, so that the bio-brick 20 can still keep good activity even in the displacement process by setting the liquid film, and the success rate of bio-printing is improved.
In addition, in this embodiment, the liquid film is formed by previously sucking the liquid for immersing the bio-brick 20 by the pipette 10, which can simplify the steps of the micro-particle manipulation method and improve the manipulation efficiency compared to the case of forming the liquid film by sucking a separate liquid outside the storage unit 30, and the liquid film formed by the liquid for immersing the bio-brick 20 has more excellent biocompatibility with the bio-brick 20 compared to the liquid film formed by other liquids, and can better maintain the bioactivity of the bio-brick 20 during the manipulation.
Moreover, in this embodiment, only one biological brick 20 is controlled in each control process, that is, in step b, only a single biological brick 20 is sucked by each pipette 10, which not only reduces the control difficulty, but also increases the success rate of control; moreover, based on this, a "dot-drawing" bio-printing mode can be realized, that is, the bio-bricks 20 can be arranged on the surface of the printing substrate 40 in a preset arrangement manner, instead of the prior art "line-drawing" continuous extrusion bio-printing mode, which requires placing the material to be printed (cell suspension containing the bio-bricks 20, adhesive, hydrogel, etc.) in a container and driving the material to be printed in a single-row linear output under the action of the enveloping flow by means of a piston or a nozzle, because the "dot-drawing" bio-printing mode can more accurately ensure the spacing, spatial position, etc. between the bio-bricks 20, and the bio-bricks 20 can be arranged more accurately according to the need, therefore, a more accurate bio-printing process can be realized, which is helpful for obtaining a more accurate biological construct, and from another perspective, it can also reduce the limitation of the bio-printing technology on the shape of the biological construct to be printed, the biological printing technology can realize the printing of more biological constructs with different shapes, the application range of the biological printing technology is effectively enlarged, and in addition, compared with the flowing cell suspension, the single biological brick 20 has better shaping capacity, so the biological brick 20 is directly controlled to carry out biological printing, the steps of carrying out subsequent shaping treatment on the cell suspension and the like can be omitted, and the accurate structure construction of the three-dimensional tissue can be more directly realized.
Furthermore, in step c of this embodiment, when the bio-brick 20 is placed at the set printing position, the liquid previously sucked into the pipette 10 still remains. Based on this, on one hand, in the placing process, part of the liquid in the suction part can be placed at the set printing position together with the particles under the action of negative pressure, and the part of the liquid is favorable for continuously keeping the bioactivity of the biological brick 20 in the printing process, and on the other hand, in the placing process, the liquid placed at the set printing position along with the biological brick 20 can be controlled not to be excessive, so that the influence of the excessive liquid outside the biological brick 20 on the positioning and biological functions of the biological brick 20 can be effectively reduced, the biological brick 20 can be positioned more accurately, and the biological brick 20 can be ensured to realize the preset biological function.
Therefore, the particle manipulation method of the present invention is applied to the field of bioprinting, and the biological brick 20 is not sucked into the interior of the pipette gun 10, but a liquid film is formed at the muzzle in advance, and the biological brick 20 is kept at the muzzle in the position contacting with the liquid film, so that the mechanical damage of the pipette gun 10 to the biological brick 20 and the pressure damage to the biological brick 20 caused by the large applied negative pressure can be effectively reduced, the biological brick 20 can be kept in a liquid environment in the whole manipulation process, good biological activity can be maintained, the biological brick 20 can be helped to maintain a preset form and good biological activity in the manipulation process, and a more accurate biological construct with better biological activity can be finally obtained; moreover, the grabbing and shifting processes of the biological bricks 20 are more stable and reliable under the action of the surface tension of the liquid film, the control efficiency and the control success rate can be effectively improved, and more accurate biological printing is realized.
Therefore, the invention also provides a bioprinting method for manipulating biological bricks as microparticles by using the microparticle manipulation method of the invention, and a biological construct prepared by using the bioprinting method of the invention.
It should be noted that, in the above embodiments, only one suction component is used to operate the microparticles, but it should be understood by those skilled in the art that a plurality of suction components may also be used to operate a plurality of microparticles simultaneously in one time, for example, in other embodiments, a plurality of pipetting guns 10 including one gun head may be used to suck a plurality of microparticles, or one or more pipetting guns 10 including a plurality of gun heads may also be used to suck a plurality of microparticles, which can further improve the operation efficiency; in other embodiments, the suction member is not limited to the embodiment of the pipette gun 10, and may be another pipette such as a pipette, a burette, or a syringe. In addition, the references to "step a" and "step b" in the present invention only indicate the sequence before and after, "step a" and "step b" may be two steps next to each other, and there may be other steps between "step a" and "step b".
The above description is only exemplary of the present invention and should not be taken as limiting the invention, as any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (15)
1. A method of manipulating particles, comprising the steps of, in order:
a, forming a liquid film at the lower end part of the suction component; and
step b: the fine particles are sucked by the sucking member having a liquid film formed on a lower end portion thereof, so that the fine particles are held on the lower end portion of the sucking member and brought into contact with the liquid film without being sucked into the lumen of the sucking member.
2. The microparticle manipulation method according to claim 1, wherein in the step a, a liquid film is formed on a lower end portion of the suction member by previously sucking a liquid by the suction member.
3. The particle manipulation method according to claim 2, wherein in the step a, the liquid previously sucked by the suction means is a liquid in which the particles are immersed.
4. The method according to claim 1, wherein in the step b, after the fine particles are sucked to a lower end portion of the suction member by the suction member, the suction operation is stopped, and the fine particles are held at the lower end portion of the suction member by a negative pressure and a tension of the liquid film.
5. The microparticle manipulation method according to claim 1, wherein in said step b, the microparticles are sucked by said sucking means having one or at least two lower end portions forming a liquid film, and each of said sucking means sucks only a single microparticle.
6. The particle manipulation method of claim 1, further comprising a step c, disposed after step b, of: and moving the suction part to be above the set placing position of the particles, and driving the particles to be separated from the lower end part of the suction part by using the suction part to be placed at the set placing position.
7. The method according to claim 6, wherein in the step c, the suction member is moved downward to bring the particles into direct contact with the set placement position, and the particles are separated from the lower end of the suction member by adhesion to the set placement position and gravity thereof and placed at the set placement position.
8. The method according to claim 6, wherein in the step c, the particles are ejected by the suction member so that the particles are separated from a lower end portion of the suction member by a positive pressure and a self gravity and are placed at the set placement position.
9. The particle manipulation method according to claim 6, wherein when the suction means places the particle at the set placement position, a part of the liquid previously sucked remains in the suction means.
10. The particle manipulation method of claim 9, further comprising a step d, disposed after step c, of: discharging the liquid previously sucked remaining in the sucking means.
11. The manipulation of particles of any one of claims 1 to 10, wherein said particles have a size of from 0.5mm to 3 mm.
12. The method of manipulating microparticles according to any one of claims 1 to 10, wherein the microparticles are gel-state microparticles capable of maintaining their own structural stability in a natural state.
13. The method of claim 12, wherein the particles are bio-bricks (20).
14. A bioprinting method, characterized in that a bio-brick (20) as said particles is manipulated by means of the particle manipulation method according to any of claims 1 to 13.
15. A biological construct prepared using the bioprinting method of claim 14.
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