A description will now be given of an injection apparatus 100 according to one embodiment of the present invention, with reference to the accompanying drawings. The injection apparatus 100 injects a predetermined material into a minute object, such as a cell and a colloid, using a capillary. The minute object spreads and can float in a fluid or liquid L. The fluid L is, for example, cell suspension and medium. Here,
The injection apparatus 100 includes, as shown in
The capillary 110 is an injection needle that is inserted into a specific region in the cell C, such as a nucleus and a cytoplasm, and injects a predetermined material, such as medication and protein, to the specific region. The capillary 110 is connectable to a loader nozzle 115, which will be described later, a plastic hollow manipulator member 116, and the pressure control mechanism 120. The capillary 110 includes, as shown in
The holder 114 has an I-shaped section, and includes a flange 114a that is connected to an O-ring 117 at one end of the manipulator member 116 and fixes and seals the injection needle 112 in its axial direction. The loader nozzle 115 is a glass tube that can be inserted into the capillary 110, and fills a predetermined material supplied from material filling means (not shown). A pressure tube 126 of the pressure control mechanism 120 is fixed onto the other end of the manipulator member 116. The manipulator member 116 is made of plastic, and has a channel 118.
The pressure control mechanism 120 controls the pressure in the capillary 110 via the manipulator member 116. The pressure control mechanism 120 includes pressure means 121a and decompression means 121b. The pressure means 121a includes a pump 122a for the positive pressure, a pressure control valve 123a that maintains the positive pressure constant, a tank 124a that applies a discharging positive pressure, a three-way selector valve 125, and a pressure tube 126. The decompression means 121b includes a pump 122b for the negative pressure, a pressure control valve 123b that maintains the negative pressure constant, a tank 124b that applies a sucking positive pressure, and a three-way selector valve 125. The pressure control mechanism 120 can apply any structure known in the art to each component, and a detailed description thereof will be omitted.
The three-way selector valve 125 is commonly used for the pressure means 121a and the decompression means 121b, and contributes to a reduction of the number of components. The three-way selector valve 125 selects one of the pressure means 121a and the decompression means 121b. The controller 182 in the control system 180 controls the selection by the three-way selector valve 125 so as to select the pressure means 121a in injecting the injection needle 112, and the decompression means 121b in cleansing the injection needle 112. This automatic switching mitigates the operator's load.
The pressure tube 126 is connected to the three-way selector valve 125 at its one end, and to the channel 118 in the manipulator member 116 at its other end. The channel 118 is connected to the injection hole 112a in the injection needle 112. The pressure tube 126 transmits the pressure of the pressure means 121a or the decompression means 121b to the channel 118 selected by the three-way selector valve 125.
The pressure means 121a is used to cleanse the injection needle 112 clogged with a material and/or cell and to inject the material.
The decompression means 121b enables the mixture gas radicals, which will be described later, to be attracted into the injection needle 112, preferably, into the whole internal surface of the injection hole 112a of the injection needle 112 just after the O-ring 117 shown in
This embodiment previously sets, through experiments and simulations, the discharge force by which the pressure means 121a injects the material into the cell C without damaging it, and the suction force by which the decompression means 121b attracts the oxygen radicals to the predetermined position.
The capillary 110 is connected to and driven by a capillary driving system 130. At least one of the dish driving system 150 that fixes and supports the dish 140 and the capillary driving system 130 has adjusting means for changing an arrangement and orientation between the capillary 110 and the cells C. The position is, for example, one or more positions in XYZ directions, and the orientation is, for example, one or more angles around each of the XYZ-axes. In this embodiment, the dish driving system 150 has an XY table that serves to move the dish 140 on an XY plane, and the capillary driving system 130 has a moving or fine adjustment function in other directions.
The capillary driving system 130 has first and second moving means, and adjusting means.
The first moving means moves the capillary 110 in a set direction (which is referred to as a “A-axis direction” or “insertion direction” in this embodiment). The first moving means includes a capillary insertion/ejection direct-acting table (A-axis table) 131 that is coupled with the capillary 110 and moves the capillary 110 in the A-axis direction.
The second moving means moves the capillary between the predetermined position and the cleansing position. The predetermined position is not specifically limited, but is an injection ready position above the dish 140 along the A-axis direction or a home position to which the capillary 110 is reset after the injection ends. In
The second moving means includes a guide shaft 132 that linearly extends in
The second moving means may use mechanical means, such as a rack and a pinion fixed onto a motor shaft, magnetic means, such as a linear motor, or other means for the guide shaft 132 and the movable part 134. A movement of the second moving means is a unidirectional linear movement in
While this embodiment uses the second moving means to moves the capillary 110 to the cleansing position, moving means for moving the cleansing means 160 may be provided to cleanse the capillary 110 at the home position or another position. In addition, both the second moving means and the moving means for the cleansing means 160 may be provided to move both the capillary 110 and the cleansing means 160. In moving the cleansing means 160, it does not have to install the whole cleansing means 160, as described later.
The adjusting means changes an arrangement and orientation between the capillary 110 and the cell C. In an illustration in
The dish 140 is a container that houses plural cells C, and has a capturing member 142 (not shown in
An amount of the fluid L is such that the cells C float in the fluid L without contacting the capturing member 142 during a normal time period and the cells C are restricted on the capturing member 142 during an attraction time period. Since the fluid L flows in the capturing member 142 from attraction ports 143, which will be described later, a sufficient amount of the fluid L needs to be supplied to the dish 140 so that the cells C can float without contacting the capturing member 142 even after the fluid L is absorbed in the capturing member 142.
The dish 140 is provided with a sensor that detects a height of the fluid L. The control system 180 supplies the fluid L to the dish 140 through the supply part (not shown) when the sensor detects that the fluid amount is lower than a predetermined height. The dish 140 may be connected to a channel that feeds the cells into which a predetermined material has been injected (or processed cells) to subsequent recovery part.
As shown in
The capturing member 142 has plural (four in
The channel 144 includes, as shown in
A more preferable embodiment replaces the channel 144 with a channel 144A that includes plural cylindrical perforations 140c, as shown in
The most preferable embodiment replaces the channel 144 with a channel 144B that includes plural cylindrical perforations 140d, as shown in
The pressure control mechanism 145 includes pressure means 145a and decompression means 145b.
The pressure means 145a includes a pump 146a for the positive pressure, a pressure control valve 147a that maintains the positive pressure constant, a tank 148a that applies a discharging positive pressure, and a three-way selector valve 149. The decompression means 145b includes a pump 146b for the negative pressure, a pressure control valve 147b that maintains the negative pressure constant, a tank 148b that applies an attracting negative pressure, and a three-way selector valve 149. The pressure control mechanism 145 has a structure similar to that of the pressure control mechanism 120, and a detailed description thereof will be omitted.
The three-way selector valve 149 selects one of channels for the pressure means 145a and the decompression means 145b. The controller 182 in the control system 180 controls the selection by the three-way selector valve 149 so as to select the pressure means 145a when the attraction port 143 is to inject, and the decompression means 145b when the attraction port 143 is to attract. This automatic switching mitigates the operator's load.
The pressure tube 141 is connected to the three-way selector valve 125 at its one end, and to the channel 144 at its other end. The channel 144 may be the above channel 144A or 144B. The pressure tube 141 transmits the pressure of the pressure means 145a or the decompression means 145b to the channel 144 selected by the three-way selector valve 149.
The pressure means 145a erupts the liquid L by pressurizing the attraction port 143, thereby releasing the cell C or separating plural aggregating cells C. The pressure means 145a is used to cleanse the attraction port 143 clogged with the material and cell(s).
The decompression means 145b can use the originally equipped decompression means that serves to fix the minute object. The decompression means 145b can attract the mixture gas radicals in the attraction port 143. This configuration can exert the cleansing effects, such as disinfection, sterilization, and wetting property, on the inner surface of the attraction port 143. The disinfection and sterilization effects prevent the contaminations in the subsequent injections. The improved wetting property secures captures of the cell C. When the decompression means 145b affects only each attraction port 143 through the channel 144A or 144B, the mixture gas radicals can be more easily attracted to the desired attraction port 143 than attractions through all the attraction ports 143.
The controller 182 of this embodiment controls the discharge force of the pressure means 145a and the suction force of the decompression means 145b. This embodiment previously sets, through experiments and simulations, the suction force used to capture the cell C without damaging the cell C, the suction force used to attract the oxygen radicals to the predetermined position, and the discharge force used to release the processed cell C. Of course, the suction force used to attract the cell C and the discharge force used to disperse the cells C with liquid L are made adjustable, thereby improving the capturing efficiency.
The dish driving system 150 serves as a means that horizontally fixes and supports the dish 140, and includes an XY table. The XY table has an X-axis table 152 and a Y-axis table 154. The X-axis table 152 moves the dish 140 in a direction parallel to the paper plane of
The observation system (not shown) includes a measuring unit that measures a shape of the cell C, and a status observing unit that observes whether the capillary 110 reaches a target position in the cell C, and whether the injection of the material ends. The measuring unit includes an optical system that acquires the cell orientation information, an image processor, an image pickup means, such as a CCD, and is implemented as a measuring unit.
The cleansing means 160 irradiates the plasma activated cleansing gas (mixture gas radicals) to the injection needle 112 and the attraction ports 143 in the dish 140. Use of common cleansing means would reduce the number of components, but the cleansing means 160 for cleansing the injection needle 112 is different from the cleansing means for cleansing the attraction ports 143 in another embodiment.
The injection apparatus 100 includes the cleansing means 160, and provides cleansing just before the injection. Although it is conceivable to cleanse the capillary 110 previously, e.g., at the shipping time from the factory, the cleansing effect does not last and the cleansing effect for improving the sterilization, disinfection, and wetting property, may be possibly lost before the first or n-th injection. The cleansing effect stabilizes the injection of the medication.
Similarly, the injection apparatus can cleanse the attraction port 143 before the cell C is attracted. If the capillary 110 is merely cleansed previously, e.g., at the shipping time from the factory, the cleansing effect for improving the sterilization, disinfection, and wetting property, may be possibly lost before the injection. The cleansing effect stabilizes capture of the cell C in the attraction port 143, and thus the subsequent injection.
The automatic, not manual, cleansing of the injection needle 112 associates with the movement of the second moving means, thereby mitigating the operator's load. A previous confirmation of the cleansing effect would eliminate uneven cleansing that could occur in the manual cleansing, and enhance the cleansing reliability. However, the present invention is not limited to the embodiment that moves the injection needle 112, but allows movements of the injection needle 112 and/or the cleansing means 160. A description will be given later of the automatic cleansing of the attraction port 143.
The cleansing means 160 includes gas supply means 161, and plasma generating means 167. When there are plural cleansing means 160, the gas supply means 161 may be shared.
The gas supply means 161 includes, as shown in
The reactive gas or process gas becomes active species due to the plasma, and serves to improve cleansing, sterilization, disinfection, and wetting property. The contaminant of the glass material of the injection needle 112 is often polymer, and oxygen radicals are combined with carbon in the polymer into CO2, providing a degradation effect. Of course, the reactive gas is not limited to oxygen gas, and may use hydrogen and other gases. The inert gas is not limited to argon, and may use other gases, such as neon, xenon, helium, radon, and krypton.
The plasma generating means 167 uses an inductively coupled plasma (“ICP”) device. The plasma generating means 167 includes a high frequency power source 168a, a matching box 168b, an inductive coil 168c, and a plasma reactor tube 168d. However, the present invention does not limit the plasma generating means to the ICP source, but may use a microwave-excited surface-wave plasma device shown in
In order to protect the injection needle 112 and the attraction port 143 from the deformations and damages due to the plasma heat, the injection needle 112 and the attraction port 143 needs to be arranged at a position that does not cause deformations by previously measuring the temperature distribution of the irradiation range IR.
Accordingly, the temperature detection system 170 detects the temperature distribution in the irradiation range IR of the oxygen radicals as the cleansing gas, and informs the temperature information to the controller 182. The temperature detecting system 170 may use, but is not limited to, any one of the temperature detecting system 170A shown in
The temperature detecting system 170A includes, as shown in
The temperature detecting system 170B attaches, as shown in
The temperature detecting system 170C attaches, as shown in
The temperature detecting systems 170A and 170B detect the temperature distribution close to that of the actual irradiation range IR, since the mixture gas that has passed the plasma reactor tube 168d spreads in all directions. Since the glass tube 174 defines the channel of the mixture gas radicals, the temperature detecting system 170C would detect the temperature distribution higher than that of the actual irradiation range IR.
The capillary 110 is mounted on an L-shaped support member 104 that can incline around the bottom end. More specifically, the support member 104 holds the holder 114. Thereafter, the capillary is cleansed at the ready position, and then the loader nozzle 115 is inserted, and the injection material is filled.
The control system 140 includes the controller 182, a memory 184, and a timer 186.
The controller 182 calculates an insertion direction and insertion position of the cell C to be inserted into the capillary 110 based on the measurement result by the measuring means in the observation system. Next, the controller 182 controls the first moving means and adjusting means in the capillary driving system 130 and driving of the dish driving system 150 based on the calculated insertion direction and the insertion position.
The controller 182 obtains temperature information of the cleansing gas (mixture gas radicals) from the temperature detecting system 170. The controller 182 controls, based on the temperature information, a distance between the injection needle 112/the attraction port 143 and the irradiating part of the cleansing gas so that the injection needle 112 and the attraction port 143 do not deform or get damaged. If necessary, the controller 182 may control the moving means and the gas flowmeter 166 of the gas supply means 161 in the cleansing means 160 and the high frequency power source 168a of the plasma generating means 167. The controller 182 controls various components of the injection apparatus 100, such as attraction/discharge parts of the cells C, supply part of the liquid L, and injecting part of the material.
The memory 142 stores operational methods of the controller 182, such as flowcharts shown in
The timer 186 calculates the predetermined time period, such as a plasma irradiation time period.
A description will be given of the cleansing method of this embodiment with reference to
Initially, the controller 182 determines the cleansing condition (step 1100).
First, the controller 182 stores the necessary heatproof characteristic information in the memory 184 (step 1102). The heatproof characteristic information contains information relating to the plasma irradiation temperature and time period that do not cause the deformations of the injection needle 112 and the material around the attraction port 143, such as glass and plastic.
Next, the controller 182 stores necessary cleansing information in the memory 184 (step 1104). The cleansing information contains a cleansing range, cleansing power, and cleansing effect. The cleansing range means, for example, the outer and internal cleansing ranges of the injection needle 112 and the dish 140, the number of attraction ports 143, and the coordinates of the attraction ports 143. The cleansing power has parameters including the negative pressure power, the plasma density, and the cleansing time period. The negative pressure power is one necessary to attract the mixture gas radicals into the predetermined range in the perforated member during cleansing. The plasma density depends upon the voltage of the high frequency power source 168a. The cleansing effect is a demanded effect to improve the sterilization, disinfection, and wetting property. Since the cleansing power usually lowers with the number of uses, the controller 182 may set the cleansing information by considering the number of uses. For example, the cleansing power for the injection needle 112 that has injected 100 times may be made several times as strong as the initial cleansing power.
Next, the controller 182 obtains the temperature distribution information of the irradiation range IR form the temperature detecting system 170 (step 1106). Finally, the controller 182 determines the irradiation time period, the irradiation position, and other irradiation conditions based on the temperature information detected by the temperature detecting system 170 (step 1108). When the controller 182 determines the irradiation position that does not cause the deformations and damages of the perforated member due to the plasma heat, the controller 182 should maintains the intended cleansing effect at that irradiation position. In this case, the controller 182 estimates the plasma density at the irradiation position, and then infers the cleansing power.
Next, the controller 182 executes cleansing in accordance with the determined cleansing condition (step 1200).
First, the controller determines whether there is a cleansing order (step 1202). The cleansing order may be an independent cleansing order, or an external input of a projection of the cell suspension, thereby automating cleansing just before the material discharge/attraction.
Next, the controller 182 moves at least one of the perforated member as a cleansing target and the cleansing means 160 via the capillary driving system 130 and the dish driving system 140 (step 1204). The cleansing position is not limited to the right position in
Next, the controller 182 determines whether the perforated member has reached the predetermined irradiation position as a result of that the at least one as the moving target has moved to the predetermined position (step 1006). The controller 182 may determine by directly detecting the perforated member through the detector (not shown) or using the information from the moving means, such as motor's rotating angle.
Next, the controller 182 activates the gas supply means 161 and plasma generating means 167 in the cleansing means 160 (step 1208), and activates the decompression means 121b or 145b (step 1210). The decompression means 145b preferably operates for each attraction port 143. Thereby, cleansing of the perforated member. Since the step 1206 arranges the perforated member at the predetermined position, the perforated member is protected from the deformations and damages due to the plasma heat. This configuration provides automatic, not manual, cleansing after the moving step 1204, mitigating the operator's load. A previous confirmation of the cleansing effect would eliminate uneven cleansing that could occur in the manual cleansing, and enhance the cleansing reliability.
Next, the controller 182 determines whether the set irradiation time period has elapsed based on the time measurement by the timer 186 (step 1212). The controller 182 continues plasma cleansing until it determines that the set irradiation time period has elapsed. When the controller 182 determines that the set irradiation time period has elapsed (step 1212), the controller 182 stops the gas supply means 161 and the plasma generating means 167 in the cleansing means 160 (step 1214), as well as stopping the decompression means 121b or 145b (step 1216).
Next, the controller 182 determines whether cleansing ends (step 1218). For example, when the perforated member is the dish 140, cleansing continues until all the attraction ports 143 are cleansed. Therefore, when the controller 182 determining that cleansing of one attraction port 143 ends returns to the step 1206, and continues the procedure by changing the irradiation position to the adjacent, untreated attraction port 143.
When the controller 182 determines that cleansing is completed (step 1218), the controller 182 moves the at least one to the original position via the capillary driving system 130 and the dish driving system 140 (step 1220).
Turning back to
The step 1300 fills the material, such as gene and medication, in the capillary 110 when the cleansing target is the injection needle 112: The step 1300 moves and adjust the capillary 110 using the first moving means and the adjusting means, and injects the material into the cell C from the injection needle 112 by controlling the pressure means 121a. The injection becomes stable due to the cleansing. The step 1300 projects the cell suspension into the dish 140 when the perforated member is the dish 140, activates the decompression means 145b, and attracts the cell C. Since the attraction becomes stable due to the cleansing, the subsequent material injection becomes stable.
Further, the present invention is not limited to these preferred embodiments, and various variations and modifications may be made without departing from the scope of the present invention. For example, this embodiment discusses the perforated member as the injection needle 112 and the dish 140, but the perforated member may be a catheter or another instrument having a pore.
Number | Date | Country | Kind |
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2006-201013 | Jul 2006 | JP | national |