JP2008035841A - Droplet-ejecting device and droplet-ejecting method - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To reduce the possibility of contamination (contamination with bacteria and pollution). <P>SOLUTION: The droplet-ejecting device 10 for ejecting a liquid containing a material acting on a sample cell toward the sample cell in a culture liquid held in a dish 11 is constituted of an ejecting head 12 holding the liquid containing the material acting on the sample cell, and having an ejecting opening for ejecting the liquid, a piezoelectric element 34 for ejecting the liquid from the ejecting opening of the ejecting head 12 according to the applied voltage, and PC (personal computer) 17 for control/image display, applying a prescribed voltage to the piezoelectric element 34 when the ejecting opening of the ejecting head 12 is present at the position in non-contact with the culture solution. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a droplet discharge device and a droplet discharge method for discharging a small amount of a liquid such as a reagent or a chemical solution.

Patent Document 1 proposes an attempt to immerse the tip of a tubular member that holds a liquid containing a reagent, a gene, or the like in a culture solution that protects the sample cell, and introduce the reagent or the like into the sample cell. .
JP 2004-321115 A

  However, in the introduction method as disclosed in Patent Document 1 above, since the tubular member holding the reagent or the like directly touches the culture solution, when sample cells in a plurality of culture vessels are processed, between the culture vessels The risk increases such as bacterial infection, and contamination (bacterial infection and pollution) such as mash of different cultures increases.

  The present invention has been made in view of the above points, and an object of the present invention is to provide a droplet discharge apparatus and a droplet discharge method that can reduce the possibility of contamination.

One aspect of the droplet discharge device of the present invention is a droplet discharge device that discharges a liquid containing a substance that acts on a sample cell toward the sample cell in the culture solution held in the container,
While holding a liquid containing a substance that acts on the sample cell, a liquid holding member having a discharge port for discharging the liquid,
Driving means for discharging the liquid from the discharge port of the liquid holding member according to the applied voltage;
Control means for applying a predetermined voltage to the driving means when the discharge port of the liquid holding member is in a position not in contact with the culture solution;
It is characterized by comprising.

Further, one aspect of the droplet discharge method of the present invention is a droplet discharge device for discharging a liquid containing a substance that acts on a sample cell toward the sample cell in the culture solution held in the container, A liquid holding member having a discharge port for discharging the liquid and holding the liquid containing a substance that acts on the sample cell, and discharging the liquid from the discharge port of the liquid holding member according to an applied voltage A droplet ejection apparatus comprising: a driving unit;
Arranging the liquid holding member at a position not in contact with the culture medium;
Discharging the liquid from the liquid holding member by the driving means when the liquid holding member is present at the position;
It is characterized by having.

  According to the present invention, there is provided a droplet discharge device and a droplet discharge method capable of reducing the possibility of contamination by discharging a liquid containing a substance that should act on a sample cell in a non-contact manner with a culture solution. Can do.

  The best mode for carrying out the present invention will be described below with reference to the drawings.

  FIG. 1 is a diagram showing a mechanical configuration of a droplet discharge device according to an embodiment of the present invention. This droplet discharge device 10 is used selectively for drug stimulation, gene introduction, fluorescent label addition (staining), etc., on any cell of a sample cell in a culture solution. (Reagent), a liquid containing a gene, a fluorescent label, or the like is discharged from a discharge head toward a desired sample cell as an object.

  The liquid droplet ejection apparatus 10 according to the present embodiment includes a dish 11, a plurality (three in FIG. 1) of ejection heads 12, piezoelectric elements (not shown in FIG. 1), an XY stage 13, a Z stage 14, It is composed of an inverted microscope 15, an imaging unit (digital camera) 16, and a control / image display personal computer (PC) 17.

  Here, the dish 11 is a container for sample cells present in the culture solution. The plurality of ejection heads 12 are liquid holding members in which different or the same ejection liquid is retained in advance, and are disposed above the dish 11 in the vertical direction. The liquid contains a drug or substance that acts on the sample cell. The piezoelectric element is a driving means that causes the inertial force to act on the liquid held by the ejection head 12 by moving the ejection head 12 back and forth in the ejection direction of the head at a high speed, thereby ejecting the liquid from the ejection head 12. is there.

  The XY stage 13 moves the dish 11 placed on the XY stage within a horizontal plane, and sets the relative position between the ejection head 12 and the dish 11 (sample) in the horizontal direction. Further, the Z stage moves along the Z rail 18 to move the ejection head 12 in the vertical direction and set the ejection position in the vertical direction.

The inverted microscope 15 acquires a microscope image of a sample using light from the light source 19 as disclosed in, for example, Japanese Patent Application Laid-Open No. 2002-174772. In the inverted microscope 15, a plurality of objective lenses 20 that are alternatively arranged in the optical path, a half mirror 21 that deflects a light beam from the light source 19 toward the objective lens 20, and an enlargement of the sample together with the objective lens 20. A primary image I _{1 of the} sample is formed by an imaging lens 22 that forms an image and two mirrors 23 and 24 that deflect the emitted light beam from the imaging lens 22 in the horizontal direction. Then, such a primary image I ₁ is relayed by the relay optical system 25 and reflected vertically upward and transmitted forward by the two mirrors 26 and 27 that can be inserted into and removed from the optical path integrally. Are selectively guided to the two light fluxes. Among these, the light beam reflected vertically upward forms an image as a _secondary image I ₂ , and further enters the eyepiece 30 attached to the lens barrel 29 after being relayed by the second relay optical system 28. It reaches the eyes of the observer and is observed. On the other hand, the forward transmitted light is guided to the imaging unit 16 via the photographing lens 31. Thus, the imaging unit 16 is an imaging unit that captures a microscopic image of the sample cell.

  The control / image display PC is connected to the inverted microscope 15 and the imaging unit 16 via cables 32 and 33, respectively, and the XY stage 13, the Z stage 14, the ejection head 12, and the like via the cables 32 and 33, respectively. The control unit includes a display that displays a microscopic image of the sample and a keyboard and a mouse as operation input units, as well as controlling the piezoelectric element and the imaging unit 16.

  In addition, as a method of filling the ejection head 12 with the sample to be ejected, the pre-filled ejection head 12 is set on the apparatus. Of course, as disclosed in JP-A-2001-235400, a Teflon (registered trademark) pipe and a syringe piston pump for suction and discharge (in addition, a Verista pump) are connected to the discharge head 12, and a sample to be discharged is A liquid supply unit for sucking in from the discharge port and discharging from the discharge port after completion of the discharge operation may be assembled in the apparatus and controlled by the control / image display PC 17. Further, the sample is often sucked from the sample cup or the like, and the sample cup may be configured on the apparatus or may be sucked by being manually immersed in the sample in the sample cup at the discharge port.

  By the way, the ejection head 12 is a head that ejects liquid by utilizing inertia as described with reference to FIG. Specifically, an inertial force acts on the liquid in the flow path by momentarily slightly moving the flow path (liquid holding member) holding the liquid containing the drug or the like, and as a result, the discharge head A part of the liquid held in the liquid 12 is ejected to the outside as droplets. This advance / retreat movement is performed by the piezoelectric element mechanically fixed to the ejection head 12 (flow path).

  FIG. 2 is a block diagram illustrating a functional configuration of the droplet discharge device 10 according to an embodiment. The control / image display PC 17 includes a control unit (PC main body) 171 that controls the entire droplet discharge device 10 and an input unit (operation unit) that performs input of settings such as a discharge position, a discharge region, and a liquid type. ) 172, a display unit 173 for inputting a setting information such as a microscope image and a discharge region, and displaying a setting result, and a storage unit 174 for storing information such as the set discharge position, discharge region, and liquid type. Prepare. Further, the storage unit 174 stores a control program executed by the control unit 171.

  In such a configuration, the control unit 171 controls the imaging unit 16 to image the sample in the dish 11 placed on the XY stage 13 through the microscope optical system of the inverted microscope 15 and obtains the image by the imaging. The displayed microscope image is displayed on the display unit 173. Then, the display unit 173 displays an input request for setting information such as a discharge region, receives an input of a discharge position, a discharge region, and a liquid type set by a user operation of the input unit 172, and displays the information on the display unit 173. And display the data in the storage unit 174. After that, according to the information stored in the storage unit 174, the XY stage 13 and the Z stage 14 are controlled to set the relative positions of the discharge head 12 and the dish 11 (sample). By applying a predetermined voltage to the piezoelectric element 34 that is fixedly fixed and moving the discharge head 12 back and forth at high speed in the discharge direction of the head, an inertial force is applied to the liquid held by the discharge head 12, thereby Thus, the liquid is ejected from the ejection head 12 to the sample in the dish 11.

  Here, an outline of the ejection head 12 will be described with reference to FIGS. 3A and 3B.

  FIG. 3A is a perspective view showing an appearance of a droplet discharge unit including the discharge head 12 and the piezoelectric element 34. As shown in the figure, the droplet discharge section uses a laminated piezoelectric element 34 as an acceleration application element. One end of the piezoelectric element 34 is fixed to the Z stage 14 as the gantry, and the other end is fixed to the ejection head 12. The discharge head 12 includes a flow path 35 and a nozzle 36. The flow path 35 includes a straight portion 35A and a tapered portion 35B. Since the piezoelectric element 34 and the flow path 35 are formed as a rigid body, the displacement of the piezoelectric element 34 does not change the volume of the flow path 35, and one end of the piezoelectric element 34 is fixed to the Z stage 14. Therefore, the entire ejection head 12 is displaced in the vertical direction in the drawing as the piezoelectric element 34 is displaced. As described above, a voltage having a desired waveform is applied to the piezoelectric element 34 from the control unit 171.

  Next, the discharge operation will be described with reference to FIG. 3B. Here, FIG. 3B is a diagram showing a temporal change with the ejection head 12 as a cross section.

  While the voltage applied from the control unit 171 to the piezoelectric element 34 is the initial voltage E0, as shown in the state (a), the droplet discharge unit is stationary. The position of the nozzle 36 at this time is A in the figure.

  When the voltage applied from the control unit 171 to the piezoelectric element 34 is gradually increased, the droplet discharge unit is slowly displaced downward in the figure as shown in the state (b) as the voltage gradually increases. To do. When the voltage applied from the control unit 171 to the piezoelectric element 34 is increased to the maximum supply voltage E1, the nozzle 36 is lowered to the position B as shown in the state (c). The maximum supply voltage E1 is a value that depends on the ejection amount, that is, the diameter of the droplet.

  Here, when the voltage applied from the control unit 171 to the piezoelectric element 34 is instantaneously reduced from the maximum supply voltage E1 to the initial voltage E0, as shown in the state (d), the droplet discharge unit also reduces the voltage. As a result, it suddenly displaces upward in the figure. At this time, the inertial force in the downward direction in the figure acts on the liquid containing the drug or the like in the flow path 35, and a relative flow is generated in the flow path 35. The flow generated in the flow path 35 is discharged from the nozzle 36 while increasing the speed at the tapered portion 35B.

  Thereafter, as shown in the state (e), the droplet discharge section returns to the initial position.

  The ejection head 12 has the following characteristics.

  The discharge port diameter of the nozzle 36 is approximately φ5 μm to φ500 μm. However, those sufficiently larger than the molecular size of the liquid to be discharged are used.

  The diameter of the droplet is in the range of φ5 μm to φ500 μm. Depending on the size of the cell, for example, the maximum supply voltage E1 to the piezoelectric element 34 is adjusted, and the diameter of the discharged droplet can be varied from the size of one cell to the size of a plurality of cells.

  The size of the droplets can be controlled by adjusting the amount of liquid present in the flow path 35 of the ejection head 12 or the driving conditions of the ejection head 12 (advance / retreat speed, voltage application time, acceleration, etc.).

  Next, the operation of the droplet discharge device 10 according to an embodiment of the above configuration will be described.

  FIG. 4A is a flowchart of a control program executed by the control unit 171 of the control / image display PC 17 of the droplet discharge device 10.

  As shown in the figure, the droplet discharge device 10 first performs an image acquisition process (step S1). This is a step of acquiring a microscopic image of the sample cell in the dish 11 by the imaging unit 16 by a known procedure and storing it in the storage unit 174.

  Next, an ejection position / liquid setting step is performed (step S2). That is, while the acquired image is displayed on the display unit 173, an input request such as the discharge position, the size (diameter) of the discharge region, and the type of liquid to be discharged is displayed. In response to this, the storage unit 174 stores discharge information such as the discharge position input from the input unit 172. That is, each of the three ejection heads 12 can eject different types of liquids, and thus allows the user to set which liquid is to be ejected and in what size.

  And an actual discharge process is implemented (step S3). In this step, the ejection information is read from the storage unit 174, and the ejection head 12 is selected, set, and ejected based on the readout information.

  When a plurality of types of liquids are ejected, the operation of the flowchart is repeated.

  Here, details of the ejection position / liquid setting process in step S2 will be described with reference to FIGS. 4B and 5A. 4B is a diagram showing a detailed flowchart of the ejection position / liquid setting step, and FIG. 5A is a diagram showing an ejection position / liquid setting screen.

  That is, as shown in FIG. 4B, in the discharge position / liquid setting step, first, an image display step is performed (step S21). This is a step of causing the display unit 173 to display the microscopic image of the sample acquired in step S1 as a determination image for determining the ejection position.

  Next, an input request process is performed (step S22). This is a step of displaying on the display unit 173 a display requesting input of information on discharge position setting.

  Thereafter, an input receiving process is performed (step S23). That is, when information such as the ejection position, the size of the ejection area, and the type of liquid to be ejected is input from the input unit 172 such as a keyboard, the input information is accepted. This is a step of accepting designation of center coordinates, diameter, and liquid type for discharging the liquid on the displayed microscope image. As a specific means for specifying the ejection area, it is preferable to use a GUI and not only a keyboard but also various input devices such as a mouse and a touch panel.

  And a discharge position display process is implemented (step S24). In this step, as shown in FIG. 5A, the information received in step S23 is superimposed on the microscope image displayed in step S21 and displayed on the display unit 173. Thereby, the operator can confirm the suitability of the discharge position. In FIG. 5A, for the sake of simplification, a microscope image is omitted, and a certain reagent (liquid), for example, reagent A (liquid A) is indicated by a solid line, and another reagent (liquid), for example, Reagent B (liquid B) is indicated by a broken line.

  As shown in the right half of FIG. 5 (A), the reagent (liquid) discharge position and the size of the discharge area are such that reagent A (liquid A) and reagent B (liquid B) are separated. It may be set, or as shown in the left half of the figure, an intersection may be made in the ejection areas of the reagents A and B, and a part of them may be set to overlap.

  Finally, a discharge position storage step is performed (step S25). That is, information such as the set ejection position is stored in the storage unit 174.

  By the way, as described in step S24 above, when setting the discharge region, adjacent discharge regions can be set so as to make a partial intersection. In this way, it is possible to form a concentration distribution in which the concentration of the reagent or substance contained in the ejected liquid is high at the intersection, and is low otherwise. That is, it is possible to set areas in which the concentration difference of the liquid to be ejected (including a reagent or substance that acts on the sample cells) is different within a very narrow range in one dish 11, and the sample in the same environment In contrast, it is possible to observe how the reagents and substances contained in the discharge liquid act depending on the concentration.

  Next, details of the ejection process in step S3 will be described with reference to FIGS. 4C, 6 and 7. FIG. FIG. 4C is a diagram showing a detailed flowchart of the ejection process, FIG. 6 is a diagram for explaining a droplet ejection system, and FIG. 7 shows an applied voltage to the piezoelectric element 34 and the ejection head 12. It is a figure which shows the relationship with forward / backward movement.

  That is, as shown in FIG. 4C, in the discharge process, first, a discharge position / discharge liquid setting readout process is performed (step S31). This is a step of reading out necessary information such as the ejection position and size stored in the storage unit 174 in step S25 from the storage unit 174.

  Next, an ejection head selection process is performed (step S32). That is, a suitable ejection head 12 is selected based on the information (liquid type to be ejected) read in step S31.

  Subsequently, an ejection head moving step is performed (step S33). This is a step of moving the ejection head 12 selected in step S32 to a position based on the information (ejection position) read in step S31. At this time, the XY stage 13 is moved with respect to the XY coordinates, and the Z stage 14 is moved with respect to the Z coordinates by an appropriate distance to secure the relative position between the selected ejection head 12 and the dish 11 (sample).

  And a drive voltage application process is implemented (step S34). That is, by applying a voltage as shown in FIG. 7 to the piezoelectric element 34 that is a driving means of the discharge head 12, the discharge head 12 moves forward and backward, thereby discharging the liquid inside the discharge head 12. . At this time, as shown in FIG. 6, the discharge position is set in the air outside the culture solution 37 in the dish 11, so that the discharge head 12 does not come into contact with the culture solution 37 in the dish 11. . And discharge is performed from the position. In the present embodiment, the voltage applied to the piezoelectric element 34 has a waveform as described in detail later.

  Thereafter, a step of determining the presence or absence of the remaining head is performed (step S35). That is, after the ejection in step S34 is completed, it is confirmed whether or not there is a remaining ejection head 12 that has not yet been ejected among the ejection heads 12 selected in step S32. If there is an ejection head 12 that has not yet ejected, the process returns to step S32, and steps S32 to S35 are repeated until all ejections are completed. And after discharge of all the selected discharge heads 12 is performed, this discharge process is ended.

  Next, the waveform of the voltage applied to the piezoelectric element 34 as the driving means of the ejection head 12 in the driving voltage applying process in step S34 will be described.

The applied voltage waveform may be a Lorentz waveform as shown in FIG. 8A, for example. As shown in the figure, this Lorentz waveform is

Defined by Here, a is the amplitude, b is the peak X-axis position, and c is FWHM (full width at half maximum).

Alternatively, the applied voltage waveform may be a Gaussian waveform as shown in FIG. 8B. This Gaussian waveform is shown in the figure,

Defined by

Furthermore, the applied voltage waveform may be a combined waveform of a Lorentz waveform and a Gaussian waveform as shown in FIG. 8C. As shown in the figure, this composite waveform is

Defined by Here, d is a correction value of the Gaussian waveform. This d takes a value of 0 ≦ d ≦ 1, and when d = 0, this combined waveform is the same waveform as the Lorentz waveform, and when d = 1, it is the same waveform as the Gaussian waveform.

  Each waveform has a steep slope, but since there is no discontinuous portion, unnecessary high-frequency vibration does not occur, and the ejection force (inertia) acting on the liquid held in the sufficient ejection head 12. Power).

  In each of the equations shown in the above formulas 1 to 3, the value of c is in the range of 1.0 μs to 5 ms and does not exceed the resonance frequency of the piezoelectric element 34. The value a is the maximum supply voltage E1 applied to the piezoelectric element 34, and the value b is half the total number of steps for operating the piezoelectric element 34.

  The piezoelectric element 34 expands when a high voltage is applied, and the expansion / contraction width of the piezoelectric element 34 is 1 μm to 100 μm. By reducing the a value of the piezoelectric element 34, a c value equal to or higher than the resonance frequency can be set. However, it is essential that the piezoelectric element 34 has no resonance.

  Note that even if a sine waveform is used as the driving voltage of the piezoelectric element 34, resonance does not occur because there is no discontinuous portion, but such a sine waveform has a gentler slope than the Gaussian waveform or the Lorentz waveform. For this reason, the range of the viscosity of the liquid that can ensure and cope with the discharge amount of the liquid is smaller than when the Gaussian waveform or Lorentz waveform is used. However, depending on the liquid used, it is of course possible to use a sine waveform.

  As described above in detail, according to the droplet discharge device 10 according to an embodiment of the present invention, a sufficient inertia force (acceleration) acts on the liquid held in the discharge head 12 to perform reliable discharge. As expected, the driving voltage of the piezoelectric element 34 is driven forward and backward by the piezoelectric element 34 by using a waveform in which the applied voltage does not change discontinuously, preferably a Gaussian waveform or Lorentz waveform, or a composite waveform thereof. Unnecessary high frequency vibration does not occur in the discharge head 12. Since high-frequency vibration does not occur in this manner, satellite droplets are not ejected, so that an appropriate amount of droplets is ejected accurately. As a result, only one droplet can reach only a desired region on the sample. That is, the droplet discharge device 10 according to the above-described embodiment can selectively and accurately cause a droplet to reach a sample in a desired region.

  In addition, when setting the areas where different liquids are discharged, it is possible to make each discharge area have an intersection. For this reason, it is possible to selectively set a region where several kinds of droplets are mixed and a region where the droplets are not mixed, that is, a region where a plurality of kinds of drugs or substances having some effect on the sample are present and a region where it is not. Therefore, the interaction of these drugs or substances with the sample can be observed.

  Furthermore, in the droplet discharge device 10 according to the above-described embodiment, the discharge port of the discharge head 12 does not touch the tip of the discharge head 12 with the liquid (culture medium 37) existing around the sample. Droplets are ejected when present in air. Therefore, since the discharge head 12 does not touch the culture solution 37 in the dish 11, even when a plurality of dishes 11 are handled, there is a dramatic risk of contamination from the dish 11 to other dishes 11 via the discharge head 12. Can be reduced.

  Although the present invention has been described above based on the embodiments, the present invention is not limited to the above-described embodiments, and various modifications and applications are naturally possible within the scope of the gist of the present invention.

  For example, in the above-described embodiment, the liquid droplet ejection apparatus 10 including the three ejection heads 12 has been described. However, the number of ejection heads 12 is not limited thereto, and may include only one. You can have as many as you like.

  5A shows an example in which two types of reagents (liquids) are discharged into two discharge regions, respectively, it is needless to say that a single region may be used instead of a plurality of regions. Alternatively, there may be no overlap by setting a plurality of areas.

  Further, the discharged liquid is not limited to a plurality of types and may be the same. In that case, as shown in FIG. 5B, by generating the intersection (overlap) of the discharge regions, a density difference from the other portions in the discharge regions can be generated. In FIG. 5B, if ideal, the density in the central portion is the highest. By producing such a concentration difference, it is possible to observe the influence of the concentration of the drug or substance in a very close region. When one kind of liquid is used as described above, the discharge may be performed once or more, and the conditions are not limited.

  Further, as shown in FIG. 5C, the liquid may be ejected so as to provide many intersections of different liquids (drugs or substances). In the example of FIG. 5C, if ideal, four types of liquids overlap in the central portion. By providing many intersections of different liquids in this way, it is possible to observe the influence of drug or substance interaction in a very close region. In addition, when using a plurality of types of liquids in this way, the discharge may be performed once or more for each liquid, and the conditions are not limited.

(Appendix)
The invention having the following configuration can be extracted from the specific embodiment.

(1) A droplet discharge device for discharging a liquid containing a substance that acts on a sample cell toward the sample cell in a culture solution held in a container,
While holding a liquid containing a substance that acts on the sample cell, a liquid holding member having a discharge port for discharging the liquid,
Driving means for discharging the liquid from the discharge port of the liquid holding member according to the applied voltage;
Control means for applying a predetermined voltage to the driving means when the discharge port of the liquid holding member is in a position not in contact with the culture solution;
A droplet discharge apparatus comprising:

(Correspondence with embodiment)
The droplet discharge device described in (1) corresponds to FIG. 1, FIG. 2, and FIG. In one embodiment, the droplet discharge device 10 is the droplet discharge device, the dish 11 is the container, the culture solution 37 is the culture solution, the discharge head 12 is the liquid holding member, and the piezoelectric element 34 is the above. The control unit 171 of the control / image display PC 17 corresponds to the drive unit, and corresponds to the control unit.

(Function and effect)
According to the droplet discharge device described in (1), in the droplet discharge device that discharges liquid toward the sample cells in the culture solution, the discharge port of the liquid holding member that discharges the liquid is immersed in the culture solution. Since the ejection is performed without the possibility of contamination, the possibility of contamination to the sample cells can be reduced.

  (2) The liquid according to (1), wherein the control means discharges the liquid by applying a voltage to the driving means when the discharge port of the liquid holding member exists in the air. Drop ejection device.

(Correspondence with embodiment)
The droplet discharge device described in (2) corresponds to FIG.

(Function and effect)
According to the droplet discharge device described in (2), the droplet is not discharged when the discharge port is in contact with the culture solution existing around the sample cell. Since it is discharged, even when handling a plurality of culture solutions, the possibility of contamination from the culture solution to other culture solutions through the discharge port can be dramatically reduced.

  (3) The drive means applies the inertial force to the liquid held by the liquid holding member and causes the liquid to be discharged from the discharge port. The droplet discharge device according to (1), wherein the droplet discharge device is advanced and retracted in a direction substantially parallel to the liquid discharge direction.

(Correspondence with embodiment)
The droplet discharge device described in (3) corresponds to FIG.

(Function and effect)
According to the droplet discharge device described in (3), discharge can be performed without worrying about clogging even if the diameter of the discharge port is small.

  (4) The control means uses, as the drive means, any one of a Gaussian waveform voltage, a Lorentz waveform voltage, and a synthesized waveform voltage of the Gaussian waveform and the Lorentz waveform as a voltage for moving the liquid holding member forward and backward. The liquid droplet ejection apparatus according to (3), wherein the liquid droplet ejection apparatus is applied.

(Correspondence with embodiment)
The droplet discharge device described in (4) corresponds to FIGS. 8A, 8B, and 8C.

(Function and effect)
According to the droplet discharge device described in (4), the voltage applied to the driving means is a voltage of a Gaussian waveform or Lorentz waveform having a steep slope but no discontinuous portion or a composite waveform thereof. Thus, unnecessary high-frequency vibration is not generated in the liquid holding member, and a sufficient discharge force (inertial force) can be obtained, so that only a desired droplet is discharged onto the object, and the satellite Generation can be suppressed.

  (5) The liquid ejecting apparatus according to (1), wherein the liquid to be ejected is a drug that stimulates the sample cell, a gene that acts on the sample cell, or a dye that stains the sample cell. .

(Function and effect)
According to the droplet discharge device described in (5), the agent that stimulates the sample cell, the gene that acts on the sample cell, or the dye that stains the sample cell can be obtained without worrying about nozzle clogging. Since it can be discharged, it can be applied to genetic testing machines and biochemical analyzers.

(6) A droplet discharge device for discharging a liquid containing a substance that acts on a sample cell toward a sample cell in a culture solution held in a container, the liquid containing the substance acting on the sample cell A liquid discharge member comprising: a liquid holding member having a discharge port that holds and discharges the liquid; and a driving unit that discharges the liquid from the discharge port of the liquid holding member according to an applied voltage. A discharge method,
Arranging the liquid holding member at a position not in contact with the culture medium;
Discharging the liquid from the liquid holding member by the driving means when the liquid holding member is present at the position;
A droplet discharge method comprising:

(Correspondence with embodiment)
The droplet discharge method described in (6) corresponds to FIG. 1, FIG. 2, FIG. 4, and FIG. In one embodiment, the droplet discharge device 10 is the droplet discharge device, the dish 11 is the container, the culture solution 37 is the culture solution, the discharge head 12 is the liquid holding member, and the piezoelectric element 34 is the above. Step S34 corresponds to the driving step and the step of arranging and the step of applying, respectively.

(Function and effect)
According to the droplet discharge method described in (6), in the droplet discharge device that discharges the liquid toward the sample cells in the culture solution, the discharge port of the liquid holding member that discharges the liquid is immersed in the culture solution. Since the ejection is performed without the possibility of contamination, the possibility of contamination to the sample cells can be reduced.

FIG. 1 is a diagram showing a mechanical configuration of a droplet discharge device according to an embodiment of the present invention. FIG. 2 is a block diagram illustrating a functional configuration of the droplet discharge device according to the embodiment. FIG. 3A is a perspective view illustrating an appearance of a droplet discharge unit including an discharge head and a piezoelectric element. FIG. 3B is a diagram showing temporal changes with the ejection head as a cross section. FIG. 4A is a diagram illustrating a flowchart of a control program executed by the control unit of the control / image display PC of the droplet discharge device according to the embodiment, and FIG. FIG. 4A is a diagram illustrating a detailed flowchart of a discharge position / liquid setting process in FIG. 4A, and FIG. 4C is a diagram illustrating a detailed flowchart of the discharge process in FIG. FIG. 5A is a diagram showing a discharge position / liquid setting screen, FIG. 5B is a diagram for explaining a plurality of superposition of a single reagent, and FIG. It is a figure for demonstrating the superimposition of a reagent. FIG. 6 is a diagram for explaining the droplet discharge method. FIG. 7 is a diagram illustrating the relationship between the voltage applied to the piezoelectric element and the forward / backward movement of the ejection head. FIG. 8A is a diagram showing a Lorentz waveform and an equation defining it. FIG. 8B is a diagram showing a Gaussian waveform and an equation defining it. FIG. 8C is a diagram showing a combined waveform of a Lorentz waveform and a Gaussian waveform and an equation defining it.

Explanation of symbols

    DESCRIPTION OF SYMBOLS 10 ... Droplet discharge apparatus, 11 ... Dish (petri dish), 12 ... Discharge head, 13 ... XY stage, 14 ... Z stage, 15 ... Inverted microscope, 16 ... Imaging part (digital camera), 17 ... Control, image Personal computer for display (PC), 18 ... Z rail, 19 ... light source, 20 ... objective lens, 21 ... half mirror, 22 ... imaging lens, 23,24,26,27 ... mirror, 25 ... relay optical system, 28 2nd relay optical system, 29 ... barrel, 30 ... eyepiece lens, 31 ... photographing lens, 32, 33 ... cable, 34 ... piezoelectric element, 35 ... flow path, 35A ... straight part, 35B ... taper part, 36 DESCRIPTION OF SYMBOLS ... Nozzle, 37 ... Culture solution, 171 ... Control part (PC main body), 172 ... Input part (operation part), 173 ... Display part, 174 ... Memory | storage part.

Claims (6)

A droplet discharge device for discharging a liquid containing a substance that acts on a sample cell toward the sample cell in a culture solution held in a container,
While holding a liquid containing a substance that acts on the sample cell, a liquid holding member having a discharge port for discharging the liquid,
Driving means for discharging the liquid from the discharge port of the liquid holding member according to the applied voltage;
Control means for applying a predetermined voltage to the driving means when the discharge port of the liquid holding member is in a position not in contact with the culture solution;
A droplet discharge apparatus comprising:   2. The droplet discharge device according to claim 1, wherein the control unit applies a voltage to the drive unit when the discharge port of the liquid holding member exists in the air, and discharges the liquid. 3. .   The driving means causes the liquid holding member to discharge the liquid according to the applied voltage in order to apply an inertial force to the liquid held by the liquid holding member and discharge the liquid from the discharge port. The droplet discharge device according to claim 1, wherein the droplet discharge device is advanced and retracted in a direction substantially parallel to the direction.   The control means applies to the driving means any one of a Gaussian waveform voltage, a Lorentz waveform voltage, and a combined waveform voltage of the Gaussian waveform and the Lorentz waveform as a voltage for moving the liquid holding member forward and backward. The droplet discharge device according to claim 3.   2. The liquid droplet ejection apparatus according to claim 1, wherein the liquid to be ejected is a drug that stimulates the sample cell, a gene that acts on the sample cell, or a dye that stains the sample cell. A droplet discharge device that discharges a liquid containing a substance that acts on a sample cell toward a sample cell in a culture solution held in a container, and holds the liquid containing the substance that acts on the sample cell And a liquid holding member having a discharge port for discharging the liquid, and a driving unit that discharges the liquid from the discharge port of the liquid holding member in accordance with an applied voltage. There,
Arranging the liquid holding member at a position not in contact with the culture medium;
Discharging the liquid from the liquid holding member by the driving means when the liquid holding member is present at the position;
A droplet discharge method comprising:

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