JP5551632B2 - Surface treatment method and surface treatment apparatus for sample manipulation element - Google Patents

Description

  The present invention treats the surface of a sheet member constituting a sample manipulation element for a sample manipulation element for performing a sealing operation in a minute space of a sample to be manipulated or a movement operation in a minute channel. The present invention relates to a surface treatment method and a surface treatment apparatus.

  For the purpose of easy handling of various trace liquids or measurement of reaction processes thereof, a micro-molded operation element for operating a trace liquid sample has been proposed. As one of such elements, a sheet member made of silicone rubber called PDMS (Polydimethylsiloxane) is used, and a sheet member having a concave or groove-like structure formed on the lower surface side is brought into close contact with the substrate. There are elements that constitute a minute sealing structure, a flow path structure, and the like (see Patent Documents 1 to 3, Non-Patent Documents 1 to 7, and the like for sample manipulation elements).

  In such an element, for example, a method is used in which a mold is manufactured with a mask or the like on which a pattern corresponding to a minute operation structure for operating a sample is formed, and a PDMS sheet member is manufactured using the mold. Can do. In this method, once a mold is made, the same element can be repeatedly produced by a simple operation and used for sample manipulation. In addition, the sheet member of PDMS has high adsorptivity to a substrate such as a glass substrate, and is suitable for creation of a microstructure by bringing the substrate and the sheet member into close contact with each other. Further, such an element can be similarly configured even when a sheet member made of a material other than PDMS is used.

JP 2004-33919 A JP 2004-309405 A JP 2005-156279 A

W. Tan and S. Takeuchi, "Atrap-and-release integrated microfluidic system for dynamic microarrayapplications", PNAS Vol.104 (2007) pp.1146-1151 S. Jo and K. Park, "Surface modification using silanated poly (ethylene glycol) s", Biomaterials 21 (2000) pp.605-616 A. Papra et al., "Microfluidic Networks Made of Poly (dimethylsiloxane), Si, and Au Coated with Polyethylene Glycol for Patterning Proteins onto Surfaces", Langmuir17 (2001) pp.4090-4095 P. Holgerson et al., "Patterning and modification of PDMS surface through laser micromachining of silicon masters and molding", Appl. Phys. A 81 (2005) pp.51-56 Hiroyuki Kudo et al., “Development of Soft Lithography Technology Using Fine Molds”, Tokyo Metropolitan Industrial Technology Research Institute Research Report No. 7 (2004) pp. 85-86 Masanori Ishizuka et al., “Assembly technology of PDMS three-dimensional structure embedded with metal electrode for microfluidic device”, IEEJ Transactions Vol. 125, No. 9 (2005) pp. 393-397 Masato Tanaka et al., “Development of chip for POCT diagnosis and laser micromachining”, Proceedings of the 73rd Laser Processing Society of Japan (2010) pp. 143-146 H. Itoh et al., "Mechanically driven ATP synthesis by F1-ATPase", Nature 427 (2004) pp.465-468

  In the sample manipulation element described above, a micro chamber (from a femtoliter (fL) to a picoliter (pL) order or a pattern of a micrometer (μm) order is formed on a PDMS by using MEMS (Micro ElectroMechanical Systems) technology. ing. Such a micromanipulation structure, for example, measures and manipulates an enzyme group called a molecular machine inside a microchamber, or uses a microchannel to discriminate molecular machines and cells and arrange them for easy measurement. , Used to manipulate various samples.

  On the other hand, when a PDMS microchamber or microchannel is used for an aqueous biological sample, the problem is that the surface of PDMS is hydrophobic. For example, when the structure of the sample manipulation element becomes finer, the aqueous solution may not enter the untreated PDMS structure. On the other hand, it has been attempted to hydrophilize the surface of the fine structure by PDMS. As such a treatment, for example, a method using an aqueous solution of an amphiphilic substance such as a surfactant can be considered. However, such a method may inhibit the function of the protein.

  Further, as a hydrophilic treatment on the surface of PDMS, there is a method in which aqueous solutions of proteins having various properties are flowed in advance or applied to the inner wall of a microchannel. Bovine serum albumin (BSA) and high-concentration aqueous solution of α-casein are candidates. In addition, physical treatment methods include a method using vacuum ultraviolet light, a method using atmospheric pressure plasma, and the like. In addition, the modification of the surface of PDMS by exposure to ozone or the like has been confirmed spectroscopically, and application examples using the reactivity have been reported (see Non-Patent Documents 2 and 3).

  The hydrophilization of PDMS by ultraviolet light irradiation treatment, plasma treatment or the like is considered to be because silanol groups are exposed on the surface of PDMS. Furthermore, there are reports that functional groups are placed or reacted with the silane coupling agent on the surface of the PDMS thus modified to use it as a functional reactive chamber or reaction channel. is there. However, these surface treatment methods basically affect the entire region of the sheet member by PDMS. For example, a specific region in an operation structure by a micro chamber, a flow path, etc. is selectively hydrophilic and functional. Until now, sufficient studies have not been made on the method of modifying the sex.

  The present invention has been made to solve the above-described problems, and a sample manipulation element constituted by a substrate and a sheet member is selectively hydrophilized in a desired region in the manipulation structure. An object of the present invention is to provide a surface treatment method and a surface treatment apparatus for a sample manipulation element that can be used.

In order to achieve such an object, a surface treatment method for a sample manipulating element according to the present invention includes a lower substrate on which a sample is placed on a placement surface, and a lower substrate on which the lower substrate is placed. An operation structure for operating a sample between the upper sheet member and the lower substrate by a concave structure provided on the lower surface of the upper sheet member. A surface treatment method for a formed sample manipulation element, wherein (1) an element preparation for preparing a sample manipulation element having a lower substrate and an upper sheet member disposed on the lower substrate and placing the sample manipulation element on a processing stage And (2) an operation structure formed between the upper sheet member and the lower substrate along the predetermined irradiation optical axis from the upper sheet member side or the lower substrate side with respect to the sample operation element on the processing stage. Beam inside Irradiating a laser beam by the condenser conditions Est is located, with plasma generated by the focusing of the laser beam, provided with a surface treatment step for hydrophilizing the inner surface of the concave structure in the upper sheet member, a surface In the processing step, a pulse laser beam is used as a laser beam, and plasma is generated by condensing the pulse laser beam in a beam waist formed in the space of the operation structure. In the surface treatment step, the sample operation element is irradiated. By adjusting the condensing position, which is the position of the beam waist of the laser beam along the optical axis, the processing conditions for the hydrophilization treatment for the position where the surface treatment is performed on the upper sheet member different from the condensing position is set. It is characterized by that.

Further, the surface treatment apparatus for a sample manipulation element according to the present invention is arranged on a lower substrate on which a sample is placed on a placement surface, and on the lower substrate so that a lower surface thereof is in close contact with the placement surface of the lower substrate. A surface with respect to the sample operation element, wherein an operation structure for operating the sample is formed between the upper sheet member and the lower substrate by a concave structure provided on the lower surface of the upper sheet member. A processing apparatus comprising: (a) a processing stage on which a sample manipulation element having a lower substrate and an upper sheet member disposed on the lower substrate is placed; and (b) a sample manipulation element on the processing stage, A laser light source for supplying laser light used for the surface treatment; and (c) an upper sheet member and a lower substrate with respect to the sample manipulating element from the upper sheet member side or the lower substrate side along a predetermined irradiation optical axis. Between A condensing optical system for irradiating a laser beam from a laser light source under a condensing condition in which a beam waist is positioned in the formed operation structure, and (d) generated by condensing the laser light in the operation structure The inside surface of the concave structure portion of the upper sheet member is hydrophilized by plasma , and the laser light source is a pulse laser light source that supplies pulse laser light as laser light, and is formed in the space of the operation structure. At the beam waist, plasma is generated by condensing pulsed laser light, and the condensing position, which is the position of the beam waist of the laser light along the irradiation optical axis with respect to the sample manipulating element, is adjusted to be different from the condensing position. and characterized in that it comprises a condensing position adjusting means for setting the processing conditions for the hydrophilization processing of a position where the surface treatment of the upper sheet member is carried out That.

  In the sample processing element surface treatment method and surface treatment apparatus described above, a sample manipulation element configured by arranging an upper sheet member made of a predetermined material such as PDMS so as to be in close contact with the placement surface of the lower substrate is provided. Prepare and place on the processing stage. Then, a laser light source and a condensing optical system are provided for the sample manipulating element on the processing stage, and a condensing condition in which the beam waist is located in the space of the manipulating structure formed between the upper sheet member and the lower substrate. Then, the laser beam is irradiated along the predetermined irradiation optical axis from the upper sheet member side or the lower substrate side.

  At this time, plasma is generated in the laser light condensing region in the operation structure of the sample operation element, and the inner surface of the concave structure portion of the upper sheet member can be hydrophilized by the laser generated plasma. In particular, in the surface treatment using the laser-generated plasma in the condensing region as described above, the substrate and the sheet member are made by appropriately setting the irradiation position of the laser beam in the horizontal plane and the condensing position in the vertical direction. It is possible to selectively perform hydrophilic treatment on the inner surface of the sheet member in a desired region of the operation structure for the sample operation element in close contact.

  Here, with respect to the laser light used for the surface treatment of the sample manipulation element, the surface treatment method preferably uses pulsed laser light as the laser light in the surface treatment step. Similarly, in the surface treatment apparatus, the laser light source is preferably a pulse laser light source that supplies pulse laser light as laser light. Thus, the hydrophilic treatment of the sheet member can be suitably realized by irradiating the sample manipulation element with the pulse laser beam.

  Further, in the surface treatment method, in the surface treatment step, the treatment position of the hydrophilic treatment for the upper sheet member is set by adjusting the irradiation position of the laser light in the plane orthogonal to the irradiation optical axis for the sample manipulation element. A configuration may be used. Similarly, the surface treatment apparatus adjusts the irradiation position of the laser beam in the plane orthogonal to the irradiation optical axis for the sample manipulation element, thereby setting the irradiation position for setting the treatment position of the hydrophilic treatment for the upper sheet member. You may use the structure provided with an adjustment means. Thereby, the irradiation position in the horizontal plane of the laser beam with respect to the sample operation element is appropriately set according to a desired region to be subjected to the hydrophilic treatment, and the upper sheet member is subjected to the hydrophilic treatment in a desired pattern. be able to.

  Further, the surface treatment method uses a configuration in which, in the surface treatment step, the processing conditions for the hydrophilic treatment for the upper sheet member are set by adjusting the condensing position of the laser light along the irradiation optical axis for the sample manipulation element. May be. Similarly, the surface treatment apparatus adjusts the condensing position of the laser beam along the irradiation optical axis with respect to the sample operating element, thereby setting the condensing position adjusting means for setting the processing conditions for the hydrophilic treatment on the upper sheet member. You may use the structure provided with. Thereby, the condensing position of the laser beam in the vertical direction with respect to the sample manipulation element can be appropriately set according to the state of the hydrophilic treatment or the like.

  Regarding the material of the sheet member that forms the upper part of the element in close contact with the mounting surface of the substrate, the material that forms the upper sheet member is preferably silicone. In particular, the material constituting the upper sheet member is preferably PDMS. By using such a material, sufficient adhesion between the substrate and the sheet member can be secured, and a sample manipulation element having a micro manipulation structure can be suitably configured.

  According to the sample processing element surface treatment method and processing apparatus of the present invention, the sample manipulation element in which the sheet member is arranged in close contact with the placement surface of the substrate is placed on the processing stage, and the sample manipulation element on the stage is placed. On the other hand, laser light is irradiated from the sheet member side or the substrate side under the light collecting condition where the beam waist is located in the operation structure between the sheet member and the substrate, and the inner surface of the sheet member is generated by laser-generated plasma. By performing the hydrophilization treatment, it becomes possible to selectively perform the hydrophilization treatment on a desired region in the operation structure of the sample operation element.

It is a figure which shows the basic composition of the sample operation element which consists of a board | substrate and a sheet | seat member. It is a schematic diagram shown about 1st Embodiment of the surface treatment method of a sample operation element. It is a figure which shows the structure of 1st Embodiment of the surface treatment apparatus of a sample operation element. It is a schematic diagram shown about 2nd Embodiment of the surface treatment method of a sample operation element. It is a figure which shows the structure of 2nd Embodiment of the surface treatment apparatus of a sample operation element. It is a figure shown about the experiment structure for evaluating the hydrophilic treatment by the surface treatment apparatus of a sample operation element. It is a figure which shows the evaluation result of the hydrophilic treatment by the surface treatment apparatus of a sample operation element. It is a figure which shows an example of the specific structure of a sample operation element. It is a figure shown about the trap of the sample in the sample operation element of FIG. It is a figure shown about an example of the surface treatment with respect to the sample operation element of FIG. It is a figure which shows the trap state of the sample in the sample operation element of FIG.

  DESCRIPTION OF EMBODIMENTS Hereinafter, preferred embodiments of a surface treatment method and a surface treatment apparatus for a sample manipulation element according to the present invention will be described in detail with reference to the drawings. In the description of the drawings, the same elements are denoted by the same reference numerals, and redundant description is omitted. Further, the dimensional ratios in the drawings do not necessarily match those described.

  The sample operation element to be surface-treated in the present invention is an element having an operation structure such as a minute sealing structure or a channel structure for the sample, and a lower part on which the sample to be operated is placed on the placement surface It is comprised by the board | substrate and the upper sheet | seat member arrange | positioned on a lower board | substrate. In addition, a micro structure such as a concave shape or a groove shape is formed on the lower surface of the sheet member, and the lower surface of the sheet member and the mounting surface of the substrate are in close contact with each other, thereby forming a sample operation structure between them. The First, the basic configuration and usage of such a sample manipulation element will be described.

  Here, in the following, a case where PDMS (Polydimethylsiloxane) is mainly used as an example of the material constituting the sheet member above the element will be described as an example. PDMS microstructures (microchannels, etc.) using MEMS technology are expected to develop industrially due to the small amount of sample, the miniaturization in combination with other devices, and the low price. Some are already in practical use.

  PDMS can transfer up to sub-micron structure by molding, it can be sealed just by attaching it to the substrate because it is self-adsorbing, and it is colorless and transparent when observing biological samples etc. using an optical microscope etc. There are advantages such as being almost free of autofluorescence, having no adverse effect on biological samples, and having no toxicity. In the bio / chemical field, there has been an attempt to develop a sample manipulation element using PDMS by providing a function to the microchannel itself. Examples are the arrangement of a three-dimensional flow path incorporating a micro pump and the enhancement of functionality by adding a partial enzyme.

  FIG. 1 is a diagram schematically showing a basic configuration of a sample manipulation element composed of a lower substrate and an upper sheet member to be processed in the surface treatment method and the surface treatment apparatus according to the present invention. Here, FIG. 1A shows the state of the sample manipulation element before the upper sheet member is brought into close contact with the lower substrate, and FIG. 1B shows the state between the upper sheet member and the lower substrate. Fig. 6 shows a state in which a sample operation element having an operation structure is formed.

  As shown in FIG. 1, the sample manipulation element 10 includes a lower substrate 12 on which a sample is placed on a placement surface 13 and an upper sheet member 14 placed on the lower substrate 12. The upper sheet member 14 is provided with a concave structure portion 16 on a lower surface 15 that is in close contact with the placement surface 13 of the lower substrate 12. Thereby, an operation structure 17 for operating the sample is formed between the sheet member 14 and the substrate 12. As described above, the sheet member 14 is made of, for example, a PDMS sheet. The substrate 12 is made of, for example, a slide glass or a glass substrate.

  FIG. 1B shows an example in which a sealing structure (a micro chamber for sealing a sample) that performs a sealing operation on a sample in a micro space is used as the operation structure 17 provided in the sample operating element 10. ing. Here, a sample solution S1 containing a target substance S2 such as a molecular machine is sealed in a space of a sealing structure 17 constituted by a concave structure portion 16 between the sheet member 14 and the substrate 12. In addition to the sealing structure described above, various structures are used as the operation structure 17 such as, for example, a flow channel structure (a micro flow channel through which a sample flows) that moves a sample in a micro flow channel. be able to.

  FIG. 2 is a schematic diagram showing the first embodiment of the surface treatment method for the sample manipulation element. In the surface treatment method according to the present embodiment, first, a sample manipulation element 10 having a lower substrate 12 and an upper sheet member 14 is prepared and placed on a processing stage (described later) (element preparation step). Then, with respect to the sample operating element 10 on the stage, the light collecting condition in which the beam waist 18 is positioned in the operating structure 17 between the sheet member 14 and the substrate 12 along the predetermined irradiation optical axis Ax from the substrate 12 side. Then, the laser beam L is irradiated. At this time, in the operation structure 17, plasma is generated by condensing the laser light L, and the inner surface of the concave structure portion 16 in the sheet member 14 is hydrophilized by the laser-generated plasma (surface treatment step).

  Here, in the following, as shown in FIG. 2, the direction (vertical direction) perpendicular to the placement surface of the substrate 12 (the lower surface of the sheet member 14) is defined as the z-axis direction, and the substrate 12 is placed perpendicular to the z-axis. A direction (horizontal direction) along the placement surface is defined as an x-axis direction and a y-axis direction. The irradiation optical axis Ax of the laser light L is set so as to substantially coincide with the z-axis direction.

  FIG. 3 is a diagram showing the configuration of the first embodiment of the surface treatment apparatus for the sample manipulation element. The configuration of the surface treatment apparatus 1A for the sample manipulation element 10 according to the present embodiment corresponds to the surface treatment method shown in FIG. A surface treatment apparatus 1A shown in FIG. 3 includes a processing stage 20, a laser light source 30, and a condensing optical system 35, and is configured as an inverted optical microscope.

  The processing stage 20 places the sample manipulation element 10 having the lower substrate 12 and the upper sheet member 14 disposed on the substrate 12. The stage 20 in this embodiment is configured by an XYZ movable stage that can move in the x-axis direction, the y-axis direction, and the z-axis direction. Thereby, the stage 20 adjusts the irradiation position of the laser beam L in the plane (xy plane) orthogonal to the irradiation optical axis Ax with respect to the sample operating element 10, thereby changing the processing position of the hydrophilic treatment on the sheet member 14. It has a function of an irradiation position adjusting means for setting.

  Further, the stage 20 adjusts the condensing position of the laser light L along the irradiation optical axis Ax (z axis) with respect to the sample operating element 10, thereby setting the processing conditions for the hydrophilic treatment for the sheet member 14. It has the function of a light collecting position adjusting means. Specifically, the stage 20 adjusts the position (see FIG. 2) of the beam waist 18 of the laser light L in the operation structure 17 of the sample operation element 10 to adjust the inside of the sheet member 14. The generation position of the laser-generated plasma in the operation structure 17 used for the hydrophilic treatment of the surface is adjusted and set.

  The laser light source 30 supplies the laser light L used for the surface treatment to the sample operating element 10 on the stage 20. As the laser light source 30, a pulse laser light source that supplies pulse laser light is preferably used. The laser beam L is a laser beam having a wavelength that passes through the substrate 12 and the sheet member 14 so that the laser beam L can be condensed in the operation structure 17. As such laser light L, for example, light (visible light) having a wavelength of 532 nm, which is a double wave from a Q-switch Nd: YAG laser light source, can be used.

  A mechanical shutter 31 and a condensing optical system 35 are provided between the laser light source 30 and the stage 20. The mechanical shutter 31 controls ON / OFF of the irradiation of the laser light L to the sample operating element 10 on the stage 20. Further, the condensing optical system 35 has a beam waist 18 in the operation structure 17 formed between the sheet member 14 and the substrate 12 along the irradiation optical axis Ax from the substrate 12 side with respect to the sample operation element 10. The laser light L from the laser light source 30 is condensed and irradiated under the condensing condition located.

  In the configuration example illustrated in FIG. 3, the condensing optical system 35 includes a lens 36, a lens 37, a dichroic mirror 39, and a condensing lens 38 in order from the laser light source 30 and the shutter 31 side. The dichroic mirror 39 reflects the pulse laser light L having a predetermined wavelength supplied from the laser light source 30 via the shutter 31 and the lenses 36 and 37 (for example, visible light having a wavelength of 532 nm described above).

  The condensing lens 38 condenses the laser light L reflected by the dichroic mirror 39 and irradiates the sample operating element 10 on the stage 20. Thereby, in the operation structure 17 of the sample operation element 10, the inner surface of the concave structure part 16 in the sheet member 14 is hydrophilized by the laser-generated plasma. As the dichroic mirror 39, for example, a mirror that reflects visible light including the laser light L and transmits near-infrared light is used.

  In the surface treatment apparatus 1A according to the present embodiment, in addition to the laser light source 30, the observation light source 40 that supplies light (540 nm or more, including near infrared light) for observing the transmission image of the sample manipulation element 10 is provided. Is provided. The observation light source 40 passes through the collimating lens 41, the near-infrared light transmission filter 42, and the condenser lens 43 from the sheet member 14 side along the irradiation optical axis Ax to the sample operation element 10 with infrared observation light (broken line). Irradiate. The observation light that has passed through the element 10 passes through the dichroic mirror 39 and reaches the CCD camera 48 via the reflection mirror 44 and the lens 45, and a transmitted image of the sample manipulation element 10 is acquired by the camera 48. Is done.

  In the surface treatment apparatus 1A, a surface treatment control unit 50 that controls the operation of each part of the apparatus is provided for the processing stage 20, the laser light source 30, and the condensing optical system 35 and the like. The control unit 50 controls the movement operation of the processing stage 20 via the stage control unit 52. Thereby, the irradiation position in the xy plane of the laser beam L with respect to the sample operation element 10 and the condensing position in the z-axis direction are controlled. Further, the control unit 50 controls the laser light supply operation of the laser light source 30 and the opening / closing operation of the mechanical shutter 31 via the laser irradiation control unit 53. When the pulsed laser beam L is emitted during plasma generation, the mechanical shutter 32 installed between the reflection mirror 44 and the lens 45 is closed to protect the observation system (CCD camera 48). As a result, the generation timing, generation conditions, and observation conditions of the laser-generated plasma for surface treatment are controlled.

  Such a surface treatment control unit 50 can be configured by a computer, for example. A display device 56 and an input device 57 are connected to the control unit 50. The display device 56 is used to display information related to the surface treatment of the sample operation element 10. Further, the input device 57 is used for inputting an instruction relating to the surface treatment conditions for the sample operating element 10 or the start and end of the surface treatment.

  The effects of the surface treatment method for the sample manipulation element 10 and the surface treatment apparatus 1A according to the present embodiment will be described.

  In the surface treatment method and the surface treatment apparatus 1A for the sample manipulation element 10 described above, the sample manipulation is configured by arranging the sheet member 14 made of a predetermined material such as PDMS so as to be in close contact with the placement surface of the substrate 12. The element 10 is prepared and placed on the processing stage 20. Then, a laser light source 30 and a condensing optical system 35 are provided for the element 10 on the stage 20, and a condensing condition in which the beam waist 18 is located in the space of the operation structure 17 between the sheet member 14 and the substrate 12. (See FIG. 2), the laser beam L is irradiated from the substrate 12 side along the irradiation optical axis Ax.

  At this time, plasma is generated in the condensing region of the laser light L in the operation structure 17, and the inner surface of the concave structure portion 16 in the sheet member 14 can be hydrophilized by the laser generated plasma. In particular, in such a surface treatment method using laser-generated plasma in the condensing region, the irradiation position in the horizontal plane of the laser light L and the condensing position in the vertical direction are set appropriately, as shown in FIG. As described above, the sample manipulation element 10 in a state where the substrate 12 and the sheet member 14 are in close contact with each other can be selectively hydrophilized on the inner surface of the sheet member 14 in a desired region of the manipulation structure 17. Is possible.

  Actually, when the pattern of the PDMS sheet member was exposed to plasma generated by condensing a pulsed laser beam having a powerful nanosecond pulse width, the hydrophilicity equivalent to a commercially available plasma processing apparatus, or a silane cup The effect of reactivity to the ring agent was obtained. Further, it has been found that the effect of hydrophilization (silanolation) lasts when the surface-treated region is immersed in water, but disappears by placing it in the air. This suggests that the effect of the surface treatment described above is not due to a modification of the surface properties due to a physical structural change but due to a chemical change.

  Here, as for the laser light L used for the surface treatment of the sample manipulation element 10, it is preferable to use a pulse laser light source for supplying the pulse laser light as the laser light source 30 as described above. In this way, by irradiating the sample manipulation element 10 with pulsed laser light, the laser beam L is emitted from the beam waist 18 formed in the space of the manipulation structure 17 between the sheet member 14 and the substrate 12. It is possible to suitably generate plasma by condensing and to suitably realize the hydrophilic treatment of the sheet member 14 by the laser-generated plasma.

Considering the plasma treatment used for surface treatment, that is, the hydrophilic effect on the surface of PDMS by diffusion of ions and electrons, it is considered that the pattern can be covered with a pulse laser of nanosecond order if the pattern formation is on the order of micrometers. . The diffusion rate of oxygen in the atmosphere is estimated to be about 10 ⁻⁶ cm ² / s. However, actually, there is a possibility that the plasma is accelerated toward the surface of the element due to the shock wave at the time of plasma generation.

  Furthermore, if it is necessary to modify the surface of the PDMS over a wide range, scanning with the laser beam L may be performed, and if it is a powerful pulse laser, a picosecond to femtosecond pulse laser beam may be used. Therefore, it is possible to form a fine hydrophilic pattern. Also, if a multi-point laser operating system such as a spatial light modulator (SLM) can withstand the use of powerful pulsed laser light, it is believed that two-dimensional simultaneous modification of the surface of the PDMS will be possible.

  Moreover, in the said embodiment, the irradiation position of the laser beam L in the surface (xy surface) orthogonal to the irradiation optical axis Ax with respect to the sample operation element 10 is adjusted with the stage 20 which functions as an irradiation position adjustment means, thereby The processing position of the hydrophilic treatment for the upper sheet member 14 is set. Thereby, the irradiation position in the horizontal plane of the laser beam L with respect to the sample operation element 10 is appropriately set or scanned according to a desired region to be subjected to the hydrophilic treatment, and desired and arbitrary for the sheet member 14. Hydrophilization treatment can be performed with this pattern.

  Moreover, in the said embodiment, the condensing position of the laser beam L along the irradiation optical axis Ax (z-axis) with respect to the sample operation element 10 is adjusted with the stage 20 which functions as a condensing position adjusting means, The treatment conditions for the hydrophilic treatment for the sheet member 14 are set. Thereby, the condensing position of the laser beam L in the vertical direction with respect to the sample operating element 10 can be appropriately set according to the status of the hydrophilic treatment or the like.

  The irradiation position adjusting unit and the light collecting position adjusting unit are realized by the XYZ movable stage 20 in the above embodiment, but are not limited to such a configuration. For example, the irradiation position adjusting unit and the condensing position adjusting unit may be realized by using the stage 20 as a fixed stage and configuring the condensing optical system 35 to be movable or adjustable. Further, both the stage 20 and the optical system 35 may be configured to be movable. In addition, these irradiation position adjusting means and condensing position adjusting means may be omitted if unnecessary.

  Silicone (silicone resin, silicone rubber) is preferably used as the material of the sheet member 14 that is in close contact with the mounting surface of the substrate 12 and constitutes the upper portion of the sample manipulation element 10. In particular, as the material of the sheet member 14, it is preferable to use the above-described PDMS that has excellent adhesion to the substrate 12. By using such a material, sufficient adhesion between the substrate 12 and the sheet member 14 can be secured, and the sample manipulation element 10 having the manipulation structure 17 can be suitably configured.

  In addition, the material of the upper sheet member 14 is not limited to the above-described silicone, PDMS, and the like, and various materials may be used as long as the minute operation structure 17 can be formed in close contact with the lower substrate 12. Examples of such materials include polydiene polymers, polyvinylidene chloride (PVDC), polyvinylidene fluoride, polyvinylidene halide, synthetic resins such as polyethylene, natural rubber, and the like.

  In addition, regarding the irradiation of the laser beam L to the sample manipulation element 10, in the embodiment shown in FIGS. 2 and 3, the configuration in which the laser beam L is irradiated from the lower substrate 12 side (lower side) is shown. It is good also as a structure which irradiates the laser beam L from the member 14 side (upper side).

  FIG. 4 is a schematic diagram showing a second embodiment of the surface treatment method for the sample manipulation element. In the surface treatment method according to the present embodiment, the beam waist is placed in the operation structure 17 between the sheet member 14 and the substrate 12 along the irradiation optical axis Ax from the sheet member 14 side with respect to the sample operation element 10 on the processing stage. The laser beam L is irradiated under the condensing condition where 18 is located. At this time, similarly to the configuration of FIG. 2, plasma is generated by condensing the laser light L in the operation structure 17, and the inner surface of the concave structure portion 16 in the sheet member 14 is hydrophilized by the laser-generated plasma. Is done.

  FIG. 5 is a diagram showing the configuration of the second embodiment of the surface treatment apparatus for the sample manipulation element. The configuration of the surface treatment apparatus 1B for the sample manipulation element 10 according to the present embodiment corresponds to the surface treatment method shown in FIG. The surface treatment apparatus 1B illustrated in FIG. 5 is similar to the surface treatment apparatus 1A illustrated in FIG. 3 in that the treatment stage 20, the laser light source 30, the condensing optical system 35, the observation light source 40, and the surface treatment control unit 50 are performed. And is configured as an upright optical microscope.

  A mechanical shutter 31 and a condensing optical system 35 are provided between the laser light source 30 and the stage 20. The mechanical shutter 31 controls ON / OFF of the irradiation of the laser light L to the sample operating element 10 on the stage 20. Further, the condensing optical system 35 has a beam waist 18 in the operation structure 17 formed between the sheet member 14 and the substrate 12 along the irradiation optical axis Ax from the sheet member 14 side with respect to the sample operation element 10. The laser light L from the laser light source 30 is condensed and irradiated under the condensing condition where is located.

  In the configuration example illustrated in FIG. 5, the condensing optical system 35 includes a lens 36, a lens 37, a dichroic mirror 39, and a condensing lens 38 in order from the laser light source 30 and the shutter 31 side. The dichroic mirror 39 reflects the pulsed laser light L supplied from the laser light source 30 via the shutter 31 and the lenses 36 and 37. The condensing lens 38 condenses the laser light L and irradiates the sample operating element 10 on the stage 20. Thereby, in the operation structure 17 of the sample operation element 10, the inner surface of the concave structure part 16 in the sheet member 14 is hydrophilized by the laser-generated plasma.

  Further, the observation light source 40 passes through the collimating lens 41, the near-infrared light transmission filter 42, the reflection mirror 46, and the condenser lens 43 from the substrate 12 side along the irradiation optical axis Ax with respect to the sample manipulation element 10. Irradiate outside observation light. Then, the observation light transmitted through the element 10 passes through the dichroic mirror 39 and reaches the CCD camera 48 via the reflection mirror 44 and the lens 45, and a transmission image of the sample manipulation element 10 is acquired by the camera 48. Is done. When the pulse laser beam L is irradiated, the mechanical shutter 32 installed between the reflection mirror 44 and the lens 45 is closed to protect the observation system (CCD camera 48).

  Next, the conditions for irradiating the sample manipulation element 10 with the laser light L will be examined. The above-described surface treatment method of the sample manipulation element 10 is to hydrophilize the inner surface of the upper sheet member 14 using PDMS or the like constituting the element 10, but as shown in FIGS. The beam waist 18 of the laser light L is formed in the space of the operation structure 17, and the position of the beam waist 18 (condensing position) is different from the position where the surface treatment is performed on the sheet member 14. . Therefore, in the above method, it is necessary to appropriately set the condensing position of the laser beam L with respect to the sheet member 14 in consideration of the generation and diffusion conditions of the plasma at the beam waist 18.

  In order to evaluate the irradiation condition of such laser light L, an experiment for evaluating the hydrophilization treatment by the surface treatment apparatus for the sample operation element was performed using the configuration shown in FIG. Here, a PDMS sheet having a thickness of 5 mm is used as the sheet member 14, and the positional relationship between the surface of the sheet member 14 to be hydrophilized and the condensing position of the laser beam L (the position of the beam waist) (particularly, In order to evaluate the distance between them, an experiment of hydrophilization treatment by irradiation with laser light L was performed on the upper surface of the sheet member instead of the inner surface of the sheet member.

  In this experiment, as shown in FIG. 6A, the configuration of an upright microscope is used, and the pulsed laser light L from the laser light source 30 is passed through the lenses 61 and 62, the dichroic mirror 63, and the objective lens 64. The top surface of the sheet member 14 is irradiated. Further, with respect to the observation light source 40, the infrared observation light from the observation light source 40 is irradiated to the sample operating element 10 including the sheet member 14 via the lens 65, the mirror 66, and the objective lens 64, and reflected from the element 10. The image is acquired by the camera 60 through the lens 64, the mirror 66, and the lens 67.

  As the laser light source 30, GCR-4 (Q-switch Nd: YAG laser) manufactured by Spectra Physics and having a maximum average output of 600 mJ (10 Hz) is used. Used as L. Moreover, the output of the laser beam L irradiated in the case of the surface treatment with respect to the sheet | seat member 14 was set, for example to 10 mJ.

  As for the evaluation of the condensing condition of the laser beam L on the sheet member 14, the stage 20 is moved by a predetermined distance in the x-axis direction and the z-axis direction as shown in FIG. The evaluation was performed by repeating the irradiation. As a specific procedure, first, the focus position F is adjusted to the upper surface of the sheet member 14 by adjusting the position in the z-axis direction of the stage 20 with respect to the focus position (condensing position) F of the laser light L generated by plasma. . Then, as shown in FIG. 6B, the focal position F by the objective lens 64 is set 50 μm above the upper surface of the sheet member 14 by adjusting the position of the stage 20 in the x-axis direction and the z-axis direction. Then, the surface treatment is performed by irradiation with the laser beam L under the condition of 10 Hz for 1 second.

  Next, the stage 20 is moved by 50 μm in the x-axis direction and the sheet member 14 is moved upward by 10 μm in the z-axis direction and approaches the focal position F. In this state, the laser beam L is irradiated under the same conditions as described above. Further, such movement of the stage 20 and irradiation of the laser beam L to the sheet member 14 are repeated. Thereby, on the upper surface of the sheet member 14, there are surface-treated surface portions in which the focal positions F move by 10 μm toward the sheet member 14 at the intervals of 50 μm in the x-axis direction in order of the measurement points 1, 2, and 3. It is formed.

  In this way, water is dropped on the surface of the PDMS by a pipette to the sheet member 14 in which the processed surface portions having different focal positions F are arranged in the moving direction of the stage, and the PDMS sheet member 14 by the low magnification objective lens. By observing the surface of the film, the state of the hydrophilic treatment was evaluated. FIG. 7 is a diagram showing an evaluation result of the hydrophilization treatment by the surface treatment apparatus for the sample manipulation element. When water is dropped on the surface of the PDMS and then the filter paper is brought closer to the side of the PDMS to suck up the water, water drops remain along the arrangement direction of the treated surface portions as shown in FIG.

  Further, when the surface of the PDMS is observed after a relatively long time (about 1 minute), as shown in FIG. 7B, the surface treated surface portion indicated with the number of the measurement point in the drawing is displayed. Depending on the position in the x-axis direction, that is, the distance between the upper surface of the sheet member 14 and the focal position F, the amount of water droplets remaining on the surface of the PDMS varies. This indicates the degree of hydrophilization treatment in each surface-treated surface portion. With reference to such a result, the preferred setting of the condensing position F of the laser beam L with respect to the sheet member 14 is described. Evaluation can be made.

  Further, when the focal position F of the laser beam L approaches the surface of the PDMS due to the scanning of the stage 20 in the z-axis direction, deformation of the PDMS and attached matter may appear. Regarding the condensing position (beam waist position) of the laser beam L with respect to the sheet member 14 to be subjected to the hydrophilization treatment, in addition to the hydrophilization conditions, such conditions are also taken into consideration in the operation structure of the sample operation element. It is preferable that the distance is set to be an appropriate distance from the surface of the PDMS.

  Next, a specific example of the surface treatment for the sample manipulation element will be described. FIG. 8 is a diagram illustrating an example of a specific configuration of the sample manipulation element 10. In FIG. 8, the planar structure pattern in the xy plane of the concave structure part 16 in the lower surface of the sheet | seat member 14, and the operation structure 17 by the sample operation element 10 by it is shown. In this embodiment, the sample operation element 10 in the case where the sample operation structure 17 has a trap type configuration is the target of the surface treatment. Here, the trap-type sample manipulation element is a micro-channel element that is devised for fixing a target substance such as a bead or a cell to a specific site in an observation field (see Non-Patent Document 1).

  In the configuration example shown in FIG. 8, the sample operation structure 17 includes a trap channel portion 83 provided between the sample solution input port 81 and the output port 82. The trap channel portion 83 is constituted by a two-stage channel portion including a first trap channel portion 84 on the input port 81 side and a second trap channel portion 85 on the output port 82 side. The first trap channel 84 is configured to flow the sample solution from the input port 81 from right to left while meandering up and down. The second trap channel portion 85 is configured to flow the sample solution from the first trap channel portion 84 from the left to the right while meandering up and down and to output to the output port 82.

  In addition to the main flow path meandering up and down, the trap flow path section 83 is provided with a plurality of shortcut flow paths 86 for performing a shortcut between adjacent main flow paths. Each of the shortcut channels 86 has a step structure including an upstream channel portion 87 having the same channel width as the main channel and a downstream channel portion 88 having a channel width narrower than the main channel. ing.

  In the sample operating element 10 having such a flow path configuration, when the sample solution S6 is flowed in the forward direction from the input port 81 to the output port 82, as schematically shown in FIGS. 9 (a) and 9 (b), a shortcut is performed. In the step portion of the flow path 86, the target substance (for example, beads) S7 in the solution S6 is trapped. In this state, when the sample solution S6 is flowed in the reverse direction from the output port 82 to the input port 81, the target substance S7 trapped in the shortcut channel 86 is released as shown in FIG. 9C. . For example, in the channel 86, if the width of the upstream channel portion 87 is 6 μm and the width of the downstream channel portion 88 is 2 μm, the target substance S7 such as beads having a size of 2 μm or more is trapped in the step portion. can do.

With respect to the sample manipulating element 10 having such a trap channel portion 83, laser light is emitted in the upstream channel portion 87 constituting the step portion of the shortcut channel 86, as schematically shown in FIG. When the surface treatment is performed by irradiation with L, only the inner surface of the PDMS sheet member at the stepped portion is selectively hydrophilized as shown by the broken line in FIG. Further, as shown in FIG. 10 (c), when the modification with the silane coupling agent, the reaction with maleimide NTA, and the Ni formation with NiCL ₂ were performed, the beads modified with His on the surface were trapped in the shortcut channel 86. We were able to.

  In this case, the beads were not released from the shortcut channel 86 even when the solution was flowed in the opposite direction. Subsequently, a high-concentration imidazole aqueous solution (500 mM) was allowed to flow through the flow path, and then the solution was allowed to flow in reverse, releasing the beads. This indicates that the stepped portion of the shortcut channel 86 is surely locally hydrophilized and activated by the laser-generated plasma generated by the irradiation with the laser beam L.

  Hereinafter, the hydrophilization process by laser generated plasma and the subsequent chemical reaction procedure will be described. First, a PDMS sheet member 14 on which an operation structure pattern is formed and a slide glass as the lower substrate 12 are prepared. The slide glass is preliminarily soaked overnight in 20NKOH, washed with MilliQ water, and dried.

  Both of these PDMS sheets and the slide glass are preliminarily exposed to plasma for 2 minutes using a plasma cleaner (manufactured by Vacuum Device Co., Ltd., Plasma Ion Bomber PIB-10 type). And if a process surface is bonded together and it processes on a hotplate at 100 degreeC for 2 minutes, both will couple | bond firmly. In the sample manipulation element 10 in which the PDMS sheet and the slide glass are thus bonded, the inside of the flow path structure created in the PDMS sheet is hydrophilic and is left in the atmosphere for several hours.

  Next, the slide glass with the PDMS sheet attached is placed on the stage of an inverted microscope (see FIG. 3), focused on a target position, and then irradiated with pulsed laser light for several seconds. Adjustment is made in advance so that the laser beam is focused and plasma is generated at a position of several μm from the surface of the PDMS pattern. Here, as the pulse laser beam L, a harmonic wave having a wavelength of 532 nm from the above-described Q-switched Nd: YAG laser was used. The effect of the laser-generated plasma at this time is almost the same as when using a plasma cleaner. When a bulk PDMS sheet is irradiated with laser light, the hydrophilicity can be clearly confirmed immediately thereafter, and if it is underwater Although hydrophilicity is maintained, hydrophilicity is lost in about 10 minutes in the atmosphere.

Here, as a specific example of surface treatment and sample manipulation, an attempt was made to construct a Ni-NTA-modified surface used for high-precision protein purification in hydrophilization (activation) by laser-generated plasma. The chelate of His to Ni is specific and becomes stronger in proportion to the number of His residues. Further, here, F ₁ -ATPase in which 10 × His-Tag is extended to the N-terminal of each of the three β subunits was used in the sample to be manipulated. When a Ni-NTA column is used for purification of His-Tag-modified (total 30 His) F ₁ -ATPase, elution is performed with a 500 mM imidazole solution. It has also been confirmed that F ₁ -ATPase does not elute at 500 mM KCL.

  The trap-type channel structure pattern in the PDMS sheet was produced with a channel width of 6 μm. The shortcut channel was designed so that the upstream channel portion had a width of 6 μm, the downstream channel portion constituting the step had a width of 2 μm, and beads having a diameter of about 4 μm could be trapped.

  First, in the PDMS flow path pattern, a region with a limited shortcut (see FIG. 10) is irradiated with a surface treatment pulse laser beam, and immediately after that MilliQ water is added to the pattern. Fill. 10 times the amount of 1% (W / W) γ-mercaptopropyltrimethoxysilane (Momentive Performance Materials, Inc., silane coupling agent TSL8380) whose pH was adjusted to 4.0 with acetic acid. Add to.

In order to prevent the operation element from drying, it is allowed to stand in a wet chamber for about 1 hour, and the flow path is washed with 10 times the amount of water, and then 2 times the amount of 10% DTT (dithiothreitol) aqueous solution is added. The SH group in the silane coupling agent is reduced. Similarly, the operating element was left in a wet chamber for 1 hour, washed with 10 times the amount of ultrapure water, and then dissolved in 10 mg / mL Maleimido-C3-NTA (Nacalai, pH 7.0 buffer). ) Is added twice and left in a wet chamber for 1 hour. Further, after washing the flow path with 10 times the amount of ultrapure water, 2 times the amount of 10 mM NiCL ₂ aqueous solution was added and left standing in the wet chamber for 20 minutes, and then the flow path was washed with 10 times the amount of ultrapure water. Then, the process ends. The sample manipulation element that has been processed is held in a wet chamber or in water until it is used.

The sample manipulation element subjected to the above treatment is placed in an optical microscope and the function of the functional group is observed. The sample used was a polystyrene bead coated with streptavidin and F ₁ -ATPase bound at a molar ratio of 500: 1. Here, for F ₁ -ATPase, His is extended by 10 residues at the C-terminals of the three β subunits. In addition, two residues obtained by modifying Cys are biotinylated near the tip of the γ subunit (see Non-Patent Document 8).

Before adding the sample to the sample manipulation element, if 5 times the amount of BSA aqueous solution of 5 to 10 mg / mL is added to the flow path in advance, the non-adherence of polystyrene beads on the wall or glass surface in the middle of the flow path. Specific binding can be prevented. Then, polystyrene beads with F ₁ -ATPase adjusted to a concentration of about ₁ pM are added to the flow path. Here, FIG. 11 is a diagram showing a sample trap state in the trap type sample manipulation element shown in FIG. In the figure, the target substance beads are visible in the flow path. When the flow rate of the solution is controlled, as shown in FIG. 11, beads are trapped one after another at the step portion of the shortcut channel.

  If the state where the water flow flows from the input port to the output port is maintained for about 10 minutes, the beads remain unreleased from the shortcut channel even if the water flow is reversed. Further, it was confirmed that the beads were not released even when 10 times the amount of 500 mM KCL was flowed from the input port side. This was the same from the output port side.

Thereafter, when 10 times the amount of 500 mM imidazole was allowed to flow from the output port side, the beads began to be released from the shortcut channel one after another after about 10 minutes. This means that the beads were specifically bound to Ni created in the shortcut channel in the channel pattern of the PDMS sheet via the His-Tag of F ₁ -ATPase. That is, this was because the laser-generated plasma activated only the shortcut part in the flow path pattern of the PDMS sheet, and the SH-silane coupling agent and then maleimide-C3-NTA reacted to chelate Ni. Is shown.

  The sample treatment element surface treatment method and the surface treatment apparatus according to the present invention are not limited to the above-described embodiments and configuration examples, and various modifications are possible. For example, various configurations other than the above may be specifically used for the configuration of the sample manipulation element to be subjected to the surface treatment. Further, a specific configuration of the surface treatment apparatus, for example, a configuration of an optical system for laser light irradiation including a laser light source and a condensing optical system, a configuration of a processing stage on which a sample operation element is placed, and the like are also shown in FIG. Various configurations other than the configuration example shown in FIG. 5 may be used.

  The present invention relates to a surface treatment method and a surface treatment for a sample manipulation element capable of selectively hydrophilizing a desired region in the manipulation structure of a specimen manipulation element constituted by a substrate and a sheet member. It can be used as a device.

DESCRIPTION OF SYMBOLS 1A, 1B ... Surface treatment apparatus of sample operation element, 10 ... Sample operation element, 12 ... Lower substrate, 13 ... Mounting surface, 14 ... Upper sheet member, 15 ... Lower surface, 16 ... Concave structure part, 17 ... Operation structure, 18 ... Beam waist,
DESCRIPTION OF SYMBOLS 20 ... Processing stage, 30 ... Laser light source, 31 ... Mechanical shutter, 32 ... Mechanical shutter, 35 ... Condensing optical system, 36, 37, 38 ... Lens, 39 ... Dichroic mirror, 40 ... Observation light source, 41 ... Collimating lens, 42 ... Near-infrared light transmission filter, 43 ... Condenser lens, 44, 46 ... Reflection mirror, 45 ... Lens, 48 ... CCD camera,
50 ... Surface treatment control unit, 52 ... Stage control unit, 53 ... Laser irradiation control unit, 56 ... Display device, 57 ... Input device,
81: input port, 82: output port, 83: trap channel, 84: first trap channel, 85: second trap channel, 86: shortcut channel, 87: upstream channel, 88 ... Downstream channel part.

Claims (8)

A lower substrate on which a sample is placed on a placement surface; and an upper sheet member arranged on the lower substrate so that a lower surface thereof is in close contact with the placement surface of the lower substrate. A surface treatment method for a sample operation element in which an operation structure for operating the sample is formed between the upper sheet member and the lower substrate by a concave structure provided on the lower surface of the member,
An element preparation step of preparing the sample manipulation element having the lower substrate and the upper sheet member disposed on the lower substrate and placing the sample manipulation element on a processing stage;
The operation formed between the upper sheet member and the lower substrate along a predetermined irradiation optical axis from the upper sheet member side or the lower substrate side with respect to the sample operation element on the processing stage. Surface treatment that irradiates a laser beam under a condensing condition in which a beam waist is located in the structure, and hydrophilizes the inner surface of the concave structure portion in the upper sheet member by plasma generated by the condensing of the laser beam With steps ,
In the surface treatment step, using the pulsed laser light as the laser light, the plasma generated by condensing the pulsed laser light is generated in the beam waist formed in the space of the operation structure,
In the surface treatment step, the upper sheet member different from the condensing position is adjusted by adjusting a condensing position that is a position of the beam waist of the laser light along the irradiation optical axis with respect to the sample operating element. A surface treatment method for a sample manipulation element, characterized in that a treatment condition for hydrophilic treatment is set for a position where the surface treatment is performed . In the surface treatment step, by adjusting the irradiation position of the laser light in a plane orthogonal to the irradiation optical axis with respect to the sample manipulation element, setting the processing position of the hydrophilic treatment for the upper sheet member the surface treatment method according to claim 1 Symbol mounting features. The surface treatment method according to claim 1 or 2, wherein the material constituting the upper sheet member is silicone. The surface treatment method according to claim 3, wherein the material constituting the upper sheet member is PDMS. A lower substrate on which a sample is placed on a placement surface; and an upper sheet member arranged on the lower substrate so that a lower surface thereof is in close contact with the placement surface of the lower substrate. A surface treatment apparatus for a sample operation element in which an operation structure for operating the sample is formed between the upper sheet member and the lower substrate by a concave structure provided on the lower surface of the member,
A processing stage on which the sample manipulation element having the lower substrate and the upper sheet member disposed on the lower substrate is placed;
A laser light source for supplying laser light used for the surface treatment to the sample manipulation element on the processing stage;
A beam waist in the operation structure formed between the upper sheet member and the lower substrate along the predetermined irradiation optical axis from the upper sheet member side or the lower substrate side with respect to the sample operation element. And a condensing optical system for irradiating the laser light from the laser light source under a condensing condition, and in the operation structure, in the upper sheet member by plasma generated by condensing the laser light While hydrophilizing the inner surface of the concave structure ,
The laser light source is a pulse laser light source that supplies pulse laser light as the laser light, and generates the plasma by condensing the pulse laser light at the beam waist formed in the space of the operation structure. Let
By adjusting the condensing position, which is the position of the beam waist of the laser beam along the irradiation optical axis with respect to the sample manipulation element, surface treatment is performed on the upper sheet member different from the condensing position. A surface treatment apparatus for a sample operation element, comprising: a light collecting position adjusting means for setting a treatment condition for a hydrophilic treatment for a position . Irradiation position adjusting means for setting the treatment position of the hydrophilic treatment on the upper sheet member by adjusting the irradiation position of the laser beam in a plane orthogonal to the irradiation optical axis with respect to the sample manipulation element. The surface treatment apparatus according to claim 5 . The surface treatment apparatus according to claim 5 or 6, wherein the material constituting the upper sheet member is silicone. The surface treatment apparatus according to claim 7, wherein a material constituting the upper sheet member is PDMS.