201014672 九、發明說明： 【發明所屬之技術領域】 本發明係關於一種微尺度孔洞結構乏製作技術， 是關於一種階段能量式微尺度孔洞結構之製造方法以 出具有精密孔徑之微尺度孔洞結構。 丨 【先前技術】 傳統用於細胞定位與抓取之微孔洞製作，多為利用微 機電（MEMS)之微小化技術，透過光微影曝光、顯影、餘^ 等基本微製程方式，製作出細胞尺度的微孔洞，然而此方法 須要昂貴之設備，且其製程繁複，耗費操:作之人力和時間。 例如美國專利第6699697號專利案中，揭露一種平面 化微電極以用以量測細胞膜離子通道之製造方法，其特徵 為_:利用光微影技術’搭配兩分子材料PDMS翻模的方式， 快速製作出微小孔洞，用以抓取細胞形成高阻抗，而量測細 胞膜表面離子通道訊號。又例如美國專利第6776896號專利 案中，揭露一種細胞定位的方法，用以量測細胞電生理訊 號，其特徵為：利用結構的設計以及灌注系統的輔助，在微 孔洞周圍形成一流場助使細胞往微孔洞方向移動，達到精確 定位細胞於微孔洞上方之目的。 又雷射鑽孔技術已被廣泛用於產業界，利用雷射激發 的高能量使材料瞬間燒融而行成孔洞，是一種方便且快速的 微小孔洞製造方式。 例如美國專利第4948941號專利案中，揭露一種雷射 7 201014672 =孔核，以製作出均勻尺寸的微孔洞，其特徵為：首先將 ^材覆蓋上犧牲層t射續孔時所形成的錐形孔洞僅於犧牲 :上’基材部分㈣句尺寸的微孔洞結構，待除去犧牲層 <即可知到均勻尺寸的微孔洞。又例如美國專利第 Γ斯號專利案中，揭露1雷射鑽孔方法，以製作出反 、形的微孔洞’其特徵為·首先如—般雷射鑽孔方法，鑽出 =錐形的微孔洞，祕㈣基材的位移及旋轉 ，使鑽孔位置 ❻ Ο 二雷射入射角度的變化’來製作出反錐形的微孔洞。又例如 美^利第7019257號專利案中，揭露-種雷射鐵孔方法， 作出—端為反錐形，—端為圓柱形㈣n其特徵 =百先般雷射觀方法，在基材上鑽㈣當大小的 洞’爾後利用基_位移及旋轉，經精密計算雷射 入射角度及位置，在基材上鑽出反錐形的微孔洞。 :般雷射鑽孔所製作出之微孔洞，其截面多為雜形或 形’特殊構型的截面形狀不易製作。在一般傳統的技術 H都是制單—雷射工作能量之方式來製作賴尺度孔 =構，若要精確控制該微尺度孔洞結構之孔徑，需要非常 4月準的雷射能量控剠及參數設計。 【發明内容】 孔，之一目的是提供—種階段能量式微尺度 H構之製造方法，以階段能量施加之方法製作㈣殊構 I且孔從精準之微尺度孔洞。 階工作能量源 為達到達到上述目的，本發明先以第 201014672 投射向一板材之第一表面之選定鑽孔位置一預定時間後，再 以第二階工作能量源投射向該微尺度孔洞。在該微尺度孔洞 中靠近該板材之第二表面被該第二階工作能量源融燒而處 於熔融態之板材以相反於該第二階工作能量源的入射方向 回融’而在該微尺度孔洞中形成一回融區.段，並在該板材之 第—表面形成·第—開孔區，且在該回融區段與該工作能量 源入射區段之交界處形成一貫通區段。 本發明採用了兩階段或兩階段以上之雷射脈波作為工 作能量源’以在一待加工的板材形成一微尺度孔洞，如此可 在該微尺度孔洞在靠近板材形成回融區段埒，可達到精密控 制該微尺度孔洞孔徑的效果。 【實施方式】 參閱第1圖所示，本發明微尺度孔洞結構的製作所使 用之裝置包括一光學操作機構1、一雷射光束調制鏡組2、 一板材3、和一工作能量源產生器4。於本實施例中，工作 能量源產生器4係為雷射產生器，但不以此為限，工作能量 源產生器4亦可以微放電加工方式所使用之技術設備來替 代，或者使用其他任何可在板材3上鑽孔、熱融之技術設 備，此時則不需使用光學操作機構1和雷射光束調制鏡組2。 光學拓作機構1包括一第一反射鏡承座11、一第二反 射鏡承座12、一第三反射鏡承座13、一第四反射鏡承座14、 一第一支架15、一第二支架16、一第三支架17和一工作平 台18。第一反射鏡承座11設置於第一支架15之底端，第 9 201014672 二反射鏡承座12設置於第一支架15之頂端’第三反射鏡承 座13設置於第三支架Π之支架連結端171 ’第四反射鏡承 座14設置於第三支架Π之承座連結侧172，第三支架17 設置於第二支架16之支架連結侧161 ’工作平台18設置於 第一支架15之工作平台連結侧151。 雷射光束調制鏡組2包括一第一反射鏡21、一第二反 射鏡22、一第三反射鏡23、〆第四反射鏡24和一聚焦鏡 25。第一反射鏡21設置於第一反射鏡承座11，第二反射鏡 22設置於第二反射鏡承座12,第三反射鏡23設置於第三反 射鏡承座13，第四反射鏡24設置於第四反射鏡承座14之 反射端141，聚焦鏡25設置於第四反射鏡承座14之聚焦端 142。 請參閱第2圖所示，利用上述之裝置，可在板材3之 選定鑽孔位置製作出一微尺度孔洞結構5。本發明方法所製 作出之微尺度孔洞結構5其孔洞周圍圓滑，且無殘污及裂 痕，可應用於細胞抓取及定位以及細胞生理訊號量測。由於 微尺度孔洞結構5的特殊構形，助使細胞與玻璃基材產生緊 密吸附，進而可量測到細胞膜表面電生理訊號。 請參閱第1至第3圖所示，首先，本發明微尺度孔洞 結構的製作方法係提供具有一預定厚度之板材3 ’其具有一 第一表面31和一第二表面32’並將其以曱醇清洗後設置於 光學操作機構1之工作平台18上。於本實施例中係選用 100〜300微米厚度之侧桂酸玻璃（b〇rosiiicate giass) ’但不以 此為限’亦可選用石英玻璃、鈉I弓玻璃（soda-lime glass)等具 201014672 有回融特性之玻璃材質，或是具有回融特性之 塑膠）。板材3之厚度可在〇〇1毫米〜1〇毫来之間塑材質（非 接著架設工作能量源產生器4並將其對準於 鏡2丨，本實施例中工作能量源產生器4係使用一反射 化碳氣體雷射，其操控需要兩種不同訊號的輪入，:之二氧 的微小脈衝訊號，主要用途為預先離子化二氧化碳，為持續 雷射可順利激發；另一為一種脈波調變 2體，使201014672 IX. Description of the Invention: [Technical Field] The present invention relates to a micro-scale hole structure fabrication technique, and relates to a phase energy micro-scale hole structure manufacturing method to have a micro-scale pore structure with a precise aperture.丨[Prior Art] The traditional micro-hole fabrication for cell localization and capture is mostly made by micro-electromechanical (MEMS) miniaturization technology, through micro-film exposure, development, and other basic micro-process methods. Cell-scale microvoids, however, this method requires expensive equipment, and its process is complicated and costly: manpower and time. For example, in U.S. Patent No. 6,699,697, a method for manufacturing a planarized microelectrode for measuring ion channel of a cell membrane is disclosed, characterized in that: _: using light lithography technology to match the two-molecule material PDMS over the mold, fast A tiny hole is made to capture the cells to form a high impedance, and the ion channel signal on the surface of the cell membrane is measured. For example, in U.S. Patent No. 6,778,896, a method of cell localization is disclosed for measuring a cell electrophysiological signal, which is characterized by the use of a structural design and the aid of a perfusion system to form a first-class field around the micro-cavity. Move the cells in the direction of the micro-cavities to precisely position the cells above the micro-holes. Laser drilling technology has been widely used in the industry. It is a convenient and fast way to manufacture tiny holes by using the high energy of laser excitation to instantly melt the material into holes. For example, in U.S. Patent No. 4,498,941, a laser 7 201014672 = hole core is disclosed to produce a micropore of uniform size, which is characterized in that: firstly, the material is covered with a sacrificial layer t to form a hole. The tapered hole is only sacrificed: the micro-hole structure of the upper part of the 'substrate part (four) sentence size, the sacrificial layer to be removed < a micropore of uniform size can be known. For example, in the U.S. Patent No. 3 patent, a laser drilling method is disclosed to produce a reverse-shaped micro-hole, which is characterized by the first method of laser drilling, drilling = cone The micro-cavities, the secret (4) substrate displacement and rotation, so that the drilling position ❻ Ο two laser incident angle changes 'to make reverse-conical micro-holes. For example, in the patent of U.S. Patent No. 7019257, a method of laser-hole method is disclosed, which is made of a reverse-tapered end, a cylindrical end (four), a characteristic=100-first laser view method, and drilled on a substrate. (4) When the size of the hole is used, the base hole_displacement and rotation are used to accurately calculate the incident angle and position of the laser, and the counter-conical micro-hole is drilled on the substrate. : The micro-holes made by the general laser drilling have a cross-section that is mostly heterogeneous or shaped. The cross-sectional shape of the special configuration is not easy to fabricate. In the general traditional technology, H is the method of making a single-laser working energy to make the scale hole = structure. To accurately control the aperture of the micro-scale hole structure, it is necessary to have a very accurate laser energy control and parameters in April. design. SUMMARY OF THE INVENTION One of the purposes of the hole is to provide a method for manufacturing a stage-energy micro-scale H-structure, which is produced by a stage energy application method (4) and a well-defined micro-scale hole. Step Working Energy Source In order to achieve the above object, the present invention first projects a selected drilling position to the first surface of a sheet for a predetermined time, and then projects the second-order working energy source toward the micro-scale hole. In the micro-scale hole, the second surface adjacent to the sheet is melted by the second-order working energy source, and the sheet in the molten state is reflowed in the incident direction opposite to the second-order working energy source' at the micro-scale A back-melting section is formed in the hole, and a first-perforation area is formed on the first surface of the plate, and a through-section is formed at a boundary between the re-melting section and the incident section of the working energy source. The invention adopts a two-stage or two-stage laser pulse wave as a working energy source to form a micro-scale hole in a plate to be processed, so that the micro-scale hole can form a remelting section near the plate. The effect of precisely controlling the micro-scale pore size can be achieved. [Embodiment] Referring to Figure 1, the apparatus for fabricating the micro-scale hole structure of the present invention comprises an optical operating mechanism 1, a laser beam modulating mirror group 2, a plate 3, and a working energy source generator 4. . In this embodiment, the working energy source generator 4 is a laser generator, but not limited thereto, the working energy source generator 4 can also be replaced by a technical device used in the micro-discharge processing mode, or use any other The technical equipment for drilling and hot-melting on the sheet 3 does not require the use of the optical operating mechanism 1 and the laser beam modulating mirror group 2. The optical extension mechanism 1 includes a first mirror holder 11, a second mirror holder 12, a third mirror holder 13, a fourth mirror holder 14, a first bracket 15, and a first Two brackets 16, a third bracket 17 and a working platform 18. The first mirror holder 11 is disposed at the bottom end of the first bracket 15, and the 9th 201014672 second mirror holder 12 is disposed at the top end of the first bracket 15. The third mirror holder 13 is disposed on the bracket of the third bracket The connecting end 171 'the fourth mirror socket 14 is disposed on the socket connecting side 172 of the third bracket ,, and the third bracket 17 is disposed on the bracket connecting side 161 of the second bracket 16 . The working platform 18 is disposed on the first bracket 15 . Work platform connection side 151. The laser beam modulation mirror group 2 includes a first mirror 21, a second mirror 22, a third mirror 23, a fourth mirror 24, and a focusing mirror 25. The first mirror 21 is disposed on the first mirror holder 11, the second mirror 22 is disposed on the second mirror holder 12, and the third mirror 23 is disposed on the third mirror holder 13 and the fourth mirror 24 The focusing end is disposed on the reflective end 141 of the fourth mirror holder 14 , and the focusing mirror 25 is disposed on the focusing end 142 of the fourth mirror holder 14 . Referring to Figure 2, a microscale hole structure 5 can be made at the selected drilling location of the sheet 3 using the apparatus described above. The micro-scale pore structure 5 made by the method of the invention has rounded pores around the pores, and has no residue and cracks, and can be applied to cell grasping and positioning and cell physiological signal measurement. Due to the special configuration of the micro-scale pore structure 5, the cell and the glass substrate are closely adsorbed, and the electrophysiological signal on the surface of the cell membrane can be measured. Referring to FIGS. 1 to 3, first, the micro-scale hole structure of the present invention is provided by providing a sheet 3' having a predetermined thickness and having a first surface 31 and a second surface 32'. The sterol is disposed on the working platform 18 of the optical operating mechanism 1 after cleaning. In the present embodiment, a glass silicate glass having a thickness of 100 to 300 μm is used, but not limited thereto. Quartz glass, soda-lime glass, etc. may be used as 201014672. Glass material with reflow characteristics, or plastic with reflow characteristics). The thickness of the plate 3 can be plasticized between 1 mm and 1 mm (the working energy source generator 4 is not erected and aligned to the mirror 2, the working energy source generator 4 in this embodiment) Using a reflective carbon gas laser, its manipulation requires the rotation of two different signals: the small pulse signal of the second oxygen, the main purpose is to pre-ionize carbon dioxide, which can be successfully excited for continuous laser; the other is a pulse Wave modulation 2 body, so that
^ 1§ (PWM^ 1§ (PWM
❹ command)’用於激發雷射，以進行鑽孔。本範例利用 5Vpp ’ 0.5%工作週率（duty Cyde)之訊號作為持續微】 衝；脈波調變信號則為400沿，5¥??，3.6%工作週率^脈 cycle)，以五個週期訊號進行鑽孔動作。 y 然後調整雷射光束調制鏡組2和工作平台18之位置 由於第四反射鏡承座14、第三支架17和工作平台18係經 由一步進馬達而連接於一電腦裝置，因此操控者可藉由電腦 裝置控制步進馬達而使工作平台18沿著第一支架15之工作 平台連結側151而於一 z軸方向往返移動，而調整聚焦鏡 25和工作平台18之間的距離，以選取適當之雷射光對焦距 離，使得雷射光聚焦於板材3之一第一表面31的聚焦點大 小在1微米〜500微米之間。 且操控者可藉由電腦裝置控制步進馬達而使第四反射 鏡承座14沿著第三支架17之承座連結側172而於一 X軸 方向往返移動，並使第三支架17沿著第二支架16之支架連 結側161而於一 Y軸方向往返移動，而於板材3之一第一 表面31選取複數個欲鑽孔位置，以製作微尺度孔洞陣列。 11 201014672 因此由本發明方法所製作 統進行整合，並可用於高微孔_列，其可與微流體系 配給，例如可本發明獨彳程序。亦可應用於流體的 等。 、構形的微尺度孔洞作為微喷頭❹ command)' is used to excite the laser for drilling. This example uses 5Vpp '0.5% duty cycle (duty Cyde) signal as continuous micro rush; pulse wave modulation signal is 400 edge, 5 ¥??, 3.6% working week rate ^ pulse cycle), to five The cycle signal performs the drilling action. y then adjusting the position of the laser beam modulating mirror group 2 and the working platform 18. Since the fourth mirror holder 14, the third bracket 17, and the working platform 18 are connected to a computer device via a stepping motor, the controller can borrow The stepping motor is controlled by the computer device to move the working platform 18 along the working platform connecting side 151 of the first bracket 15 in a z-axis direction, and the distance between the focusing mirror 25 and the working platform 18 is adjusted to select an appropriate The laser light is focused at a distance such that the focus of the laser light focused on one of the first surfaces 31 of the sheet 3 is between 1 micrometer and 500 micrometers. And the controller can control the stepping motor by the computer device to reciprocate the fourth mirror holder 14 along the bearing coupling side 172 of the third bracket 17 in an X-axis direction, and the third bracket 17 is along The bracket of the second bracket 16 is coupled to the side 161 to reciprocate in a Y-axis direction, and a plurality of positions to be drilled are selected on one of the first surfaces 31 of the sheet 3 to form a micro-scale array of holes. 11 201014672 is thus integrated by the method of the present invention and can be used in a high microporous column, which can be dispensed with a microfluidic system, such as the unique procedure of the present invention. It can also be applied to fluids, etc. Micro-scale holes of configuration as micro-sprinklers
❹ 選取好雷射光束調制鏡組2和工作平台18之位置後， 即可開始進仃鑽孔由於本實施例中之工作能量源產生器* 係為雷射產生器，因此藉由雷射激發訊號的操控，激發工作 能量源產生ϋ 4發射-工作能量源4卜預定時間，工作能 量源41係為具有一預定脈衝數、脈衝寬度、脈衝頻率之雷 射光束。工作能量源41會經由第一反射鏡21反射至第二反 射鏡22，再經由第二反射鏡22反射至第三反射鏡23，再經 由第三反射鏡23反射至第四反射鏡24,再經由第四反射鏡 - .j 24反射至聚焦鏡25，最後經由聚焦鏡25之聚焦而成為聚焦 後之雷射光束42且投射向板材3之第一表面31之選定鑽孔 位置。 然後聚焦後之雷射光束42開始融燒板材3之第一表面 31之選定鑽孔位置之板材，而在板材3之第/表面31形成 一第一開孔區51，而後鑽入板材3中而在鄰近板材3之第 一表面形成一工作能量源入射區段54，直到聚焦後之雷射 光束42恰穿透板材3之第二表面32,而形成/貫穿第一表 面31和第二表面32之微尺度孔洞5。 之後，請參閱第4圖所示’在微尺度孔同5中’靠近 板材3之第二表面32被聚焦後之雷射光束42棘燒而處於溶 融態之板材會以相反於聚焦後之雷射光束42的入射方向回 12 201014672 流一小段距離而凝固，而在微尺度孔洞5中靠近板材3之第 二表面處形成一回融區段55，並在板材3之第二表面32形 成一第二開孔區52，且在回融區段55與工作能量源入射區 段54之交界處（或說第一開孔區51與第二開孔區52之交界 處）形成一貫通區段53，回融區段55係自第二開孔區52延 伸連通至貫通區段53，且回融區段55之孔徑大小係自第二 開孔區52漸縮至貫通區段53。 工作能量源入射區段54係自第一開孔區51延伸連通 © 至貫通區段53，且工作能量源入射區段54之孔徑大小係自 第一開孔區51漸縮至貫通區段53。第二開孔區52係對準 連通於第一開孔區51。貫通區段53係鄰近於第二開孔區 52。第一開孔區51具有一第一開孔孔徑dl，第二開孔區52 5 具有一第二開孔孔徑d2,貫通區段53具有一貫通孔徑d3， 貫通孔徑d3係小於第一開孔孔徑dl，貫通孔徑d3係小於 第二開孔孔徑d2。且貫通區段53之位置較靠近第二開孔區 52,因此在上述回融現象後所形成之微尺度孔洞5係呈不對 ❹稱沙漏狀。 於本實施例中所製作出之第二開孔孔徑d2之大小與一 般細胞尺度相近，約為20〜30微米，適合用於細胞抓取與定 位；而貫通孔徑d3之大小與傳統片膜箝制技術(patch-clamp technique)所使用的微針電極（micropipette)尺寸相符，約為 2〜10微米，可用於研究細胞膜表面離子通道電生理反應。 此一特殊構型可順利引導細胞位於微尺度孔洞5之第一開 孔區51，提升細胞與玻璃間的緊密電阻(seal resistance)，甚 13 201014672 至達到ίο9歐姆以上，而可順利量測細胞表面離子進出的微 小電流。 又，本發明方法所製作之貫通孔徑id3係可在500奈米 〜200微米之間。而若欲使用厚度在0.01毫米〜10毫米之間 之板材3製作介於500奈米〜200微米之間之貫通孔徑r3， 需使用上述預定能量之工作能量源41(本實施例中預定能量 之工作能量源41係為預定雷射能量之雷射光束），且使其發 射一預定時間，影響此預定雷射能量之參數包含其脈衝數、 參 脈衝寬度和脈衝頻率，例如其脈衝數係可選在1〜101G之間， 而其脈衝寬度和脈衝頻率則隨之而調整，且所照射之時間亦 可由操作者視板材3之厚度、材質和所欲#作之微孔洞大小 而調整。至於所使用之工作能量源產生器彳4除可使用氣體式 .1 雷射光束產生器外，亦可使用固體式雷射光束產生器或半導 體雷射光束產生器，且其亦可為脈衝式雷射光束產生器或連 續式雷射光束產生器。 請參閱第5圖所示，茲配合前述實施例之圖式對本發 ® 明之整個操作流程作一說明。在本發明實施例，採用了兩階 段式雷射脈波作為工作能量源，以在一待加工的板材形成一 微尺度孔洞。 第6圖係本發明在製作微尺度孔洞結構時，採用兩階 段式雷射脈波作為工作能量源之第一實施例波形示意圖。兩 階段式雷射脈波包括有第一階雷射脈波S1及第二階雷射脈 波S2，其中第一階雷射脈波S1中具有複數個序列脈波P1; 第二階雷射脈波S2中具有複數個序列脈波P2,且第二階雷 14 201014672 射脈波S2之序列脈波P2之脈波寬度比第一階雷射脈波SI 之序列脈波P1之脈波寬度為窄。亦即，該第二階雷射脈波 S2之工作能量與第一階雷射脈波S1之工作能量為不同。本 實施例中，該第二階雷射脈波S2之工作能量較第一階雷射 脈波S1之工作能量為小。 同時參閱第6圖及第7圖。首先，提供具有一預定厚 度之板材（步驟101)，其具有一第一表面和一第二表面。然 後將板材設置於一工作平台上（步驟102)。 之後，再以工作能量源產生器發射一具有第一階工作 能量之第一階雷射脈波S1作為第一階工作能量源一預定時 間（步驟103)，並使第一階工作能量源經由一雷射光束調制 鏡組投射向板材之第一表面之選定鑽孔位置（步驟104)。 .i 第一階工作能量源會融燒板材之第一表面之選定鑽孔 位置之板材，並在鄰近板材之第一表面形成一工作能量源入 射區段，直到第一階工作能量源恰穿透板材之第二表面（步 驟105)，因而在板材之第一表面之選定鑽孔位置形成一貫 穿板材之第一表面和第二表面之微尺度孔洞（步驟106)。 接著，施加一具有第二階工作能量之第二階雷射脈波 S2作為第二階工作能量源至微尺度孔洞。此時，靠近板材 之第二表面被第二階工作能量源融燒而處於炼融態之板材 以相反於第二階工作能量源的入射方向回融（步驟107)，因 而在微尺度孔洞中靠近板材之第二表面處形成一回融區 段，並在板材之第二表面形成一第二開孔區，且在回融區段 與工作能量源入射區段之交界處形成一貫通區段（步驟 15 201014672 108)，回融區段係自第二開孔區延伸連通至貫通區段，且其 孔徑係自第二開孔區漸縮至貫通區段。 由於第二階雷射脈波S2之工作能量較第一階雷射脈波 S1之工作能量為小，故可在該微尺度孔洞在靠近板材之第 二表面形成回融區段時，可達到精密控制孔徑的效果。 前述步驟105中，工作能量源產生器所產生的第一階 雷射脈波S1係在恰穿透板材之第二表面而在板材之選定鑽 孔位置形成微尺度孔洞（步驟106)之後，改換施加第二階雷 射脈波S2作為工作能量源至微尺度孔洞而形成回融區段。 本發明亦可選擇其它等效的作法，同樣能達到相似的功能。 例如工作能量源產生器所產生的第一階雷射脈波S1亦可在 將近穿透板材之第二表面之前，改換為第二階雷射脈波S2 作為工作能量源施加至微尺度孔洞而形成回融區段。 第7圖係本發明在製作微尺度孔洞結構時，採用兩階 段式雷射脈波作為工作能量源之第二實施例波形示意圖。在 此實施例中，第一階雷射脈波S1中之各個序列脈波P1與 第二階雷射脈波S2中之各個序列脈波P2之脈波寬度相 同，但第二階雷射脈波S2之頻率比第一階雷射脈波S1之 頻率為小。 第8圖係本發明在製作微尺度孔洞結構時，採用兩階 段式雷射脈波作為工作能量源之第三實施例波形示意圖。在 此實施例中，第一階雷射脈波S1中之各個序列脈波P1之 脈波寬度比第二階雷射脈波S2中之各個序列脈波P2之脈 波寬度為大，但第二階雷射脈波S2之頻率比第一階雷射脈 16 201014672 、 · 波S1之頻率為南。 、第9圖係本^明在製作微尺度孔洞結構時，採用雨階 &式雷射脈波作為作能量源之第四實施例波形示意圖。在 此實施例中’第階雷射脈波S1中之各個序列脈波之 脈波寬度相同於第二階雷射脈波S2中之各個序列脈波P2 之脈波寬度，且兩者頻率也相同，但第二階雷射脈波§2中 之各個序列脈波P2之脈波高度比第—階雷射脈波si中之 各個序列脈波P1之脈波高度為低。 ® 以上之實施例係以兩階段式工作能量作為實施例説 明本發明亦可依實際需要而改變其工作能量之階數(例如 三階段或更多階段）。此外，為達到不同大小之工作能量， 除了使用脈波蛘之階段能量型態之外，當然亦可採用漸變武 之工作能量（例如線性降低式工作能量）。 由以上之實施例可知，本發明所提供之階段能量式微 尺度孔洞結構之製造方法確具產業上之利用價值，故本發明 業已符合於專利之要件。惟以上之敘述僅為本發明之較佳實 ® 施例說明，凡精於此項技藝者當可依據上述之說明而作其它 種種之改良，惟這些改變仍屬於本發明之發明精神及以下所 界定之專利範圍中。 【囷式簡單說明】 第1圖係本發明微尺度孔洞結構的製作方法及其使用的裳 置示意圖； 第2圖係顯示在一板材開設有一微尺度孔洞結構的立體圖； 17 201014672 第3圖係本發明微尺度孔洞結構的製作方法中工作能量源 融燒一板材之剖視圖； 第4圖係本發明微尺度孔洞結構之剖視圖； 第5圖係本發明微尺度孔洞結構的製作方法之流程圖； 第6圖係本發明在製作微尺度孔洞結構時，採用兩階段式雷 射脈波作為工作能量源之第一實施例波形示意圖； 第7圖係本發明在製作微尺度孔洞結構時，採用兩階段式雷 射脈波作為工作能量源之第二實施例波形示意圖； © 第8圖係本發明在製作微尺度孔洞結構時，採用兩階段式雷 射脈波作為工作能量源之第三實施例波形示意圖； 第9圖係本發明在製作微尺度孔洞結構時，採用兩階段式雷 射脈波作為工作能量源之第四實施例波形示意圖。 .1 【主要元件符號說明】： 1 光學操作機構 11 第一反射鏡承座 12 第二反射鏡承座 13 第三反射鏡承座 14 第四反射鏡承座 141 反射端 142 聚焦端 15 第一支架 151 平台連結側 16 第二支架 18 201014672 ❹ ❹ 161 支架連結侧 17 第三支架 171 支架連結端 172 承座連結侧 18 工作平台 2 雷射光束調制鏡組 21 第一反射鏡 22 第二反射鏡 23 第三反射鏡 24 第四反射鏡 25 聚焦鏡 3 板材 31 第一表面 32 第二表面 4 工作能量源產生器 41 工作能量源 42 聚焦後之雷射光束 5 微尺度孔洞 51 第一開孔區 52 第二開孔區 53 貫通區段 54 工作能量源入射區段 55 回融區段 dl 第一開孔孔徑 19 201014672 d2 第二開孔孔徑 d3 貫通孔徑 X X軸方向 Y Y軸方向 Z Z轴方向 51 第一階雷射脈波 P1 第一階雷射脈波之序列脈波 52 第二階雷射脈波选取 After selecting the position of the laser beam modulation mirror group 2 and the working platform 18, the drilling operation can be started. Since the working energy source generator* in this embodiment is a laser generator, the laser excitation is performed. The operation of the signal excites the working energy source to generate ϋ 4 the emission-working energy source 4 for a predetermined time. The working energy source 41 is a laser beam having a predetermined pulse number, pulse width, and pulse frequency. The working energy source 41 is reflected to the second mirror 22 via the first mirror 21, reflected to the third mirror 23 via the second mirror 22, and then reflected to the fourth mirror 24 via the third mirror 23, and then It is reflected by the fourth mirror -.j 24 to the focusing mirror 25, and finally focused by the focusing mirror 25 to become the focused laser beam 42 and projected to the selected drilling position of the first surface 31 of the sheet 3. The focused laser beam 42 then begins to melt the plate at the selected drilling location of the first surface 31 of the sheet 3, and a first apertured region 51 is formed in the first/surface 31 of the sheet 3 and then drilled into the sheet 3. A working energy source incident section 54 is formed adjacent to the first surface of the sheet 3 until the focused laser beam 42 penetrates the second surface 32 of the sheet 3 to form/through the first surface 31 and the second surface. 32 micro-scale holes 5. After that, please refer to the picture 4 in the micro-scale hole and 5 in the vicinity of the second surface 32 of the plate 3 after being focused, the laser beam 42 is spinnated and the plate in the molten state will be opposite to the focused thunder. The incident direction of the beam 42 is solidified by a short distance of 12 201014672, and a remelting section 55 is formed in the micro-scale hole 5 near the second surface of the sheet 3, and a second surface 32 is formed on the second surface 32 of the sheet 3. a second opening area 52, and a through section is formed at the interface between the back-melting section 55 and the working energy source incident section 54 (or the junction of the first opening area 51 and the second opening area 52) The remelting section 55 extends from the second opening area 52 to the through section 53 , and the aperture size of the remelting section 55 is tapered from the second opening area 52 to the through section 53 . The working energy source incident section 54 extends from the first opening area 51 to the through section 53, and the aperture size of the working energy source incident section 54 is tapered from the first opening area 51 to the through section 53. . The second opening area 52 is aligned to communicate with the first opening area 51. The through section 53 is adjacent to the second opening area 52. The first opening area 51 has a first opening diameter d1, the second opening area 52 5 has a second opening diameter d2, and the through section 53 has a through hole d3, and the through hole d3 is smaller than the first opening. The aperture dl, the through aperture d3 is smaller than the second aperture aperture d2. Moreover, the position of the through section 53 is closer to the second opening area 52, so that the micro-scale holes 5 formed after the above-mentioned remelting phenomenon are not nicknamed hourglass. The second aperture diameter d2 produced in this embodiment is similar to the general cell scale, about 20 to 30 micrometers, and is suitable for cell grasping and positioning; and the size of the through-hole diameter d3 is different from the conventional film clamping. The micropipette used in the patch-clamp technique is about 2 to 10 microns in size and can be used to study ion channel electrophysiological reactions on the cell membrane surface. This special configuration can smoothly guide the cells in the first opening area 51 of the micro-scale hole 5, and improve the sealing resistance between the cells and the glass, even 13 201014672 to reach ίο9 ohm or more, and can smoothly measure the cells. A small current that enters and exits surface ions. Further, the through-hole id3 produced by the method of the present invention may be between 500 nm and 200 m. If a platen 3 having a thickness of between 0.01 mm and 10 mm is to be used to form a through-hole r3 between 500 nm and 200 m, the working energy source 41 of the predetermined energy is used (the predetermined energy in this embodiment). The working energy source 41 is a laser beam of predetermined laser energy and is emitted for a predetermined time. The parameters affecting the predetermined laser energy include the number of pulses, the pulse width and the pulse frequency, for example, the number of pulses thereof. It is selected between 1 and 101G, and its pulse width and pulse frequency are adjusted accordingly, and the time of illumination can be adjusted by the operator depending on the thickness, material and size of the micro-holes. As for the working energy source generator 彳4, in addition to the gas type .1 laser beam generator, a solid laser beam generator or a semiconductor laser beam generator may be used, and it may also be pulsed. Laser beam generator or continuous laser beam generator. Referring to Figure 5, the entire operation of the present invention will be described in conjunction with the drawings of the foregoing embodiments. In the embodiment of the present invention, a two-stage laser pulse wave is used as a working energy source to form a micro-scale hole in a sheet to be processed. Fig. 6 is a waveform diagram showing the first embodiment of the present invention in which a two-stage laser pulse wave is used as a working energy source in the fabrication of a micro-scale hole structure. The two-stage laser pulse wave includes a first-order laser pulse wave S1 and a second-order laser pulse wave S2, wherein the first-order laser pulse wave S1 has a plurality of sequence pulse waves P1; the second-order laser wave The pulse wave S2 has a plurality of sequence pulse waves P2, and the pulse width of the sequence pulse wave P2 of the second order Ray 14 201014672 jet pulse wave S2 is larger than the pulse wave width of the sequence pulse wave P1 of the first order laser pulse wave SI It is narrow. That is, the operating energy of the second-order laser pulse S2 is different from the operating energy of the first-order laser pulse S1. In this embodiment, the working energy of the second-order laser pulse wave S2 is smaller than the working energy of the first-order laser pulse wave S1. See also Figures 6 and 7. First, a sheet having a predetermined thickness (step 101) is provided having a first surface and a second surface. The sheet is then placed on a work platform (step 102). Thereafter, the first-order laser pulse S1 having the first-order working energy is emitted as the first-order working energy source by the working energy source generator for a predetermined time (step 103), and the first-order working energy source is passed through A laser beam modulation mirror is projected onto a selected drilling location of the first surface of the panel (step 104). .i a first-stage working energy source that melts the sheet of the selected drilling location on the first surface of the sheet and forms a working energy source incident section adjacent the first surface of the sheet until the first stage working energy source is worn The second surface of the sheet is passed through (step 105), thereby forming a micro-scale hole through the first and second surfaces of the sheet at selected locations of the first surface of the sheet (step 106). Next, a second-order laser pulse S2 having a second-order working energy is applied as a second-order working energy source to the micro-scale holes. At this time, the second surface adjacent to the sheet is melted by the second-order working energy source, and the sheet in the fused state is melted back in the incident direction opposite to the second-order working energy source (step 107), thus being in the micro-scale hole Forming a remelting section near the second surface of the sheet, and forming a second opening area on the second surface of the sheet, and forming a through section at the boundary between the remelting section and the incident section of the working energy source (Step 15 201014672 108), the remelting section extends from the second opening zone to the through section, and the aperture is tapered from the second opening zone to the through section. Since the working energy of the second-order laser pulse wave S2 is smaller than the working energy of the first-order laser pulse wave S1, the micro-scale hole can be formed when the remelting section is formed near the second surface of the plate. Precision control of the aperture effect. In the foregoing step 105, the first-order laser pulse S1 generated by the working energy source generator is changed after the micro-scale hole is formed at the selected drilling position of the plate just after penetrating the second surface of the plate (step 106). A second order laser pulse S2 is applied as a source of working energy to the micro-scale holes to form a remelting section. The invention may also select other equivalents to achieve similar functions. For example, the first-order laser pulse S1 generated by the working energy source generator may be changed to the second-order laser pulse wave S2 as a working energy source to be applied to the micro-scale hole before passing through the second surface of the plate. Form a remelting section. Fig. 7 is a waveform diagram showing a second embodiment of the present invention in which a two-stage laser pulse wave is used as a working energy source in the fabrication of a micro-scale hole structure. In this embodiment, the pulse wave widths of the respective sequence pulse waves P1 in the first-order laser pulse wave S1 and the respective sequence pulse waves P2 in the second-order laser pulse wave S2 are the same, but the second-order laser pulse The frequency of the wave S2 is smaller than the frequency of the first-order laser pulse S1. Fig. 8 is a waveform diagram showing a third embodiment of the present invention in which a two-stage laser pulse wave is used as a working energy source in the fabrication of a micro-scale hole structure. In this embodiment, the pulse width of each of the sequence pulse waves P1 in the first-order laser pulse wave S1 is larger than the pulse width of each of the sequence pulse waves P2 in the second-order laser pulse wave S2, but The frequency of the second-order laser pulse S2 is higher than the first-order laser pulse 16 201014672, and the frequency of the wave S1 is south. The ninth figure is a waveform diagram of the fourth embodiment in which the rain pulse & type laser pulse wave is used as the energy source when the micro-scale hole structure is fabricated. In this embodiment, the pulse width of each of the sequence pulse waves in the first-order laser pulse wave S1 is the same as the pulse wave width of each of the sequence pulse waves P2 in the second-order laser pulse wave S2, and the frequencies of the two are also The same, but the pulse height of each of the sequence pulse waves P2 in the second-order laser pulse § 2 is lower than the pulse wave height of each of the sequence pulse waves P1 in the first-order laser pulse si. The above embodiments use two-stage working energy as an example to illustrate that the present invention can also change the order of its working energy (e.g., three stages or more) as needed. In addition, in order to achieve different working energy, in addition to the phase energy pattern of the pulse wave, it is of course also possible to use the working energy of the gradient (for example, linear reduced working energy). It can be seen from the above embodiments that the manufacturing method of the stage energy type micro-scale hole structure provided by the present invention has industrial utilization value, and therefore the invention has met the requirements of the patent. However, the above description is only a description of the preferred embodiment of the present invention, and those skilled in the art can make other improvements according to the above description, but these changes still belong to the inventive spirit of the present invention and the following Within the scope of the defined patent. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a schematic view showing a method for fabricating a micro-scale pore structure of the present invention and a skirt for use thereof; and FIG. 2 is a perspective view showing a micro-scale pore structure in a sheet; 17 201014672 FIG. FIG. 4 is a cross-sectional view showing a micro-scale hole structure of the present invention; FIG. 5 is a flow chart showing a method for fabricating the micro-scale hole structure of the present invention; Figure 6 is a waveform diagram of a first embodiment of the present invention for producing a micro-scale hole structure using a two-stage laser pulse wave as a working energy source; Figure 7 is a second embodiment of the present invention for producing a micro-scale hole structure. A waveform diagram of a second embodiment of a staged laser pulse as a working energy source; © FIG. 8 is a third embodiment of the present invention for producing a micro-scale hole structure using a two-stage laser pulse wave as a working energy source Waveform diagram; FIG. 9 is a fourth embodiment of the present invention using a two-stage laser pulse wave as a working energy source when fabricating a micro-scale hole structure Schematic diagram. .1 [Description of main component symbols]: 1 Optical operating mechanism 11 First mirror holder 12 Second mirror holder 13 Third mirror holder 14 Fourth mirror holder 141 Reflecting end 142 Focusing end 15 First Bracket 151 Platform joint side 16 Second bracket 18 201014672 ❹ 161 161 Bracket joint side 17 Third bracket 171 Bracket joint end 172 Seat joint side 18 Working platform 2 Laser beam modulation mirror group 21 First mirror 22 Second mirror 23 Third mirror 24 Fourth mirror 25 Focusing mirror 3 Plate 31 First surface 32 Second surface 4 Working energy source generator 41 Working energy source 42 Focused laser beam 5 Micro-scale hole 51 First opening area 52 second opening area 53 through section 54 working energy source incident section 55 backing section dl first opening aperture 19 201014672 d2 second opening aperture d3 through aperture XX axis direction YY axis direction ZZ axis direction 51 First-order laser pulse wave P1 first-order laser pulse sequence pulse wave 52 second-order laser pulse wave
P 2 第二階雷射脈波之序列脈波P 2 second-order laser pulse sequence pulse wave
2020