200918897 九、發明說明： 【發明所屬之技術領域】 本發明係關於一種具溫度自補償功能之微反應槽，更 進一步地，本發明之係關於一種具溫度自補償功能之核酸 增幅晶片。 【先前技術】 生物晶片並未有明碟的定義與分類，一般而言係指以 石夕晶片、玻璃或高分子為基材，配合微小化技術整合微機 電、光電、化學、生化、醫學工程及分子生物學等領域， 用以執行醫療檢驗、環境檢測、食品檢驗、新藥開發、基 礎研究、軍事防禦、化學合成等用途的精密微小化設備。 目前市面上將生物晶片分為基因晶片（gene chip )、蛋白質 晶片（protein chip )及縮微實驗室晶片（lab-on-a-chip )三 大類。其中縮微實驗室晶片的功能則依不同的需求進行設 計，在微晶片上進行不同的反應，目前已知可在微縮實驗 室晶片上進行的生化實驗包括具有基因增幅功能的聚合酶 鏈鎖反應（polymerase chain reaction，PCR)、核酸的定序 反應、微流體操作、電泳（electrophoresis )、質譜分析（MS， mass spectrograph)、抗原-抗體結合或一般的酵素反應等。 基於現有之核酸增幅晶片技術，主要加熱器設計均為 片狀或是蛇行狀金屬薄膜，在加熱時會因均勻性不足導致 核酸增幅時的專一性以及複製效率降低。本發明嘗試開發 一種加熱溫度均勻之加熱器態樣，藉此使晶片上的溫度均 勻性提高，使核酸增幅時的專一性及複製效率提高。 200918897 【發明内容】 有鑑於習知技術的缺失，本發明之目的在開發一種微 反應槽，具有加熱溫度均勻之加熱器型態，將其應用於生 物晶片，特別是對溫控有精確需求之核酸增幅晶片，可提 高加熱之均勻性。 為達上述目的，本發明之微反應槽，其係包含：一腔 室，該腔室之内部具有一表面；及一陣列型加熱器，其係 位於前述腔室表面，該陣列型加熱器係由一第一加熱層及 一第二加熱層結合形成，其中前述第一加熱層係由複數個 同軸相之條狀微加熱器構成，第二加熱層係由佈局於前1述 第一加熱層之微加熱器上以陣列形式排列之微加熱器構 成。 本發明同時也包含一種核酸增幅晶片，其特徵在於該 晶片上具有一上述之微反應槽。 本發明之微反應槽可藉由熱的自補償效應提高熱均勻 性，毋須外接溫度補償元件，且易於應用在晶片，特別是 核酸增幅晶片上，提高核酸增幅的專一性及複製效率。 【實施方式】 本發明之微反應槽，其係包含：一腔室，該腔室之内 部具有一表面；及一陣列型加熱器。陣列型加熱器之型態 係可參考第一圖所示，該陣列型加熱器其係位於前述腔室 表面1上，由一第一加熱層2及一第二加熱層3結合形成， 由圖中可知，第一加熱層2係由複數個同轴相之條狀微加 200918897 熱器2’構成，而第二加熱層3則是由佈局於前述條狀微加 熱器2’上以陣列形式排列之微加熱器3’構成。 當可輕易理解而無須限定的是，本發明之微反應槽係 可由一開口向上之槽體與一基板結合形成，或者由一開口 向下之槽體與一基板結合形成，而本發明之微反應槽中之 陣列型加熱器係可設置於前述槽體之表面或者基板之表 面，使槽體與基板結合後形成腔室而令陣列型加熱器位於 該腔室中。 形成腔室之槽體與基板之材質包含玻璃、石英或高分 子材料等樣品可於其中進行加熱反應，特別是聚合酶鏈反 應而穩定之材質，高分子枕料則包含，但不限於壓克力 (PMMA)、聚碳酸酯（PC)或聚二曱矽氧烷（PDMS) 等。 本發明之微反應槽主要應用於微型晶片系統中，適用 於微量樣品反應之用，因此微反應槽之自身體積以及設置 於其中之各元件係為微/奈米尺寸。因此微反應槽中之陣列 型微加熱器係可利用微機電技術或微影對為技術進行製 作，用以構裝成如第一圖所示之陣列型微加熱器。參考第 一圖，一般而言，條狀微加熱器之寬度d與次一條狀微加 熱器間之間隔距離d’之比例係為0.1〜10之間，取決於系統 所使用的散熱裝置、微加熱器材料以及晶片基材的種類， 另外，為了要達到自補償的效果，每一單一微加熱器之尺 寸均可有所調整，在較佳的實施態樣中，第一層之條狀微 加熱器中最外側之微加熱器寬度係為中間微加熱器寬度之 2倍以上。 200918897 本發明之反應槽其中之陣列型微加熱器的製作方法係 可利用任何微機電技術之步驟來達成，首先可在腔室表面 1上形成片狀微加熱器作為第一加熱層2 ’之後利用微機電 技術將片狀微加熱器切割成同軸向的條狀微加熱器2，，戋 者利用光阻剝落法（lift_off)法直接於腔室表面i上形成 同軸向的條狀微加熱器2,，然後再以微影對位技術形^第 一加熱層3，其中第二加熱層3係如圖中所示佈局於條狀 微加熱器2’上以陣列形式排列成微加熱器3,之態樣。 ，一般而言’微反應槽中之第一加熱層及第二加熱層所 ，之微加熱器需具備高導熱性及/或高導電度之特性，因此 多f用=屬材料，本發明所稱之金屬材料包含專_一金屬材 料或者合金材料，例如導熱性佳且安定耐用之鉑/鉻金 銘/鈦金屬。 =較於傳統片狀或蛇行狀之金屬加熱層容易因均句性 之反應二，酸增幅時的專一性以及複製效率降低，本發明BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a microreactor having a temperature self-compensation function, and more particularly to a nucleic acid amplification wafer having a temperature self-compensation function. [Prior Art] Biofilm does not have the definition and classification of open discs. Generally speaking, it refers to the integration of micro-electromechanical, optoelectronic, chemical, biochemical, medical engineering with micro-chemical technology based on Shixi wafer, glass or polymer. In the fields of molecular biology, such as precision micro-devices for medical inspection, environmental testing, food testing, new drug development, basic research, military defense, and chemical synthesis. Biochips are currently classified into three types: gene chips, protein chips, and lab-on-a-chips. The functions of the micro-laboratory wafers are designed according to different requirements, and different reactions are performed on the micro-wafers. Biochemical experiments that are currently known to be performed on micro-laboratory wafers include polymerase chain reaction with gene amplification function ( Polymerase chain reaction, PCR), sequencing of nucleic acids, microfluidic manipulation, electrophoresis, mass spectrometry, antigen-antibody binding, or general enzyme reactions. Based on the existing nucleic acid amplification wafer technology, the main heater design is a sheet-like or serpentine metal film, which causes a decrease in specificity of nucleic acid amplification and a decrease in replication efficiency due to insufficient uniformity upon heating. The present inventors have attempted to develop a heater pattern having a uniform heating temperature, thereby increasing the uniformity of temperature on the wafer and improving the specificity and replication efficiency of nucleic acid amplification. In view of the lack of the prior art, the object of the present invention is to develop a micro-reaction tank having a heater type with uniform heating temperature, which is applied to a bio-chip, especially for temperature control. The nucleic acid amplifies the wafer to improve the uniformity of heating. To achieve the above object, the microreactor of the present invention comprises: a chamber having a surface inside; and an array type heater located on the surface of the chamber, the array type heater Formed by a combination of a first heating layer and a second heating layer, wherein the first heating layer is composed of a plurality of strip-shaped micro-heaters of a coaxial phase, and the second heating layer is disposed by the first heating layer of the first one. The microheater is composed of microheaters arranged in an array. The invention also encompasses a nucleic acid amplification wafer characterized by having a microreaction cell as described above. The microreactor of the present invention can improve the thermal uniformity by the thermal self-compensation effect, without external temperature compensating components, and is easy to be applied to wafers, especially nucleic acid amplifying wafers, to improve the specificity and replication efficiency of nucleic acid amplification. [Embodiment] The microreactor of the present invention comprises: a chamber having a surface inside; and an array type heater. The type of the array heater can be referred to the first figure. The array heater is located on the surface 1 of the chamber, and is formed by combining a first heating layer 2 and a second heating layer 3. It can be seen that the first heating layer 2 is composed of a plurality of coaxial phase strips plus a 200918897 heater 2', and the second heating layer 3 is arranged in an array on the strip-shaped microheater 2'. The arranged microheaters 3' are constructed. It can be easily understood that, without limitation, the micro-reaction tank of the present invention can be formed by combining an open-up groove body with a substrate, or by forming an open-down groove body and a substrate, and the present invention The array type heater in the reaction tank can be disposed on the surface of the tank body or the surface of the substrate, and the tank body is combined with the substrate to form a chamber in which the array type heater is placed. The material of the cavity and the substrate forming the chamber includes a sample in which a sample such as glass, quartz or a polymer material can be heated, in particular, a polymerase chain reaction and a stable material, and the polymer pillow includes, but is not limited to, a press. Force (PMMA), polycarbonate (PC) or polydioxane (PDMS). The microreactor of the present invention is mainly used in a microchip system and is suitable for a small sample reaction. Therefore, the volume of the microreactor itself and the components disposed therein are micro/nano size. Thus, the array type microheater in the microreactor can be fabricated using MEMS or lithography techniques to form an array type microheater as shown in the first figure. Referring to the first figure, in general, the ratio of the width d of the strip-shaped micro-heater to the distance d' between the sub-micro-heaters is between 0.1 and 10, depending on the heat sink used in the system, The heater material and the type of the wafer substrate, in addition, in order to achieve the self-compensation effect, the size of each single micro-heater can be adjusted. In a preferred embodiment, the strip of the first layer is slightly The width of the outermost microheater in the heater is more than twice the width of the intermediate microheater. 200918897 In the reaction tank of the present invention, the method of fabricating the array type microheater can be achieved by using any microelectromechanical technology step. First, a sheet-like microheater can be formed on the chamber surface 1 as the first heating layer 2'. The micro-electromechanical technology is used to cut the micro-heater into a strip-shaped micro-heater 2 in the same axial direction, and the latter uses a lift-off method to form a strip-shaped micro-heater in the same axial direction directly on the chamber surface i. 2, and then in the form of a micro-image alignment technology, the first heating layer 3, wherein the second heating layer 3 is arranged on the strip-shaped micro-heater 2' as shown in the figure as an array of micro-heaters 3 , the way. Generally speaking, the micro-heater of the first heating layer and the second heating layer in the micro-reaction tank is required to have high thermal conductivity and/or high conductivity, so that the multi-f is a material, and the present invention The metal material is called a special metal material or alloy material, such as platinum/chromium gold/titanium with good thermal conductivity and stability. = The metal heating layer of the conventional sheet or serpentine is easily reacted by the uniformity of the second sentence, the specificity of the acid amplification and the reduction of the replication efficiency, the present invention
自補i二，微加熱器透過陣列式之設計達到熱均勻性之 圍與四彻三圖所示，其係為第一圖之俯視圖，圖中外 之調整而羞=因陣列式之設計致使可進行部分區域溫度場 場一致（熱均^溫度補償之效果，進而達到加熱區域溫度 ‘、、=勻性提高）之目的。 之生物晶片微反應槽可應用於例如需進行微量樣品加熱 種具有舒、+， 特別疋核酸增幅晶片，因此本發明包含一 當可^微反應槽之核酸增幅晶片。 反應；中二易ΐ解的是’本發明之核酸増幅晶片所設置之 '、了刖述陣列型微加熱器外，可進一步包含一溫 200918897 度感測器，用以精確量測反應槽内溫度，進而達到溫控目 的以應用於需精密溫控之聚合酶鏈反應。 本發明之核酸增幅晶片之材料係可相同或不同於設置 於其上之微反應槽之材質，關於微反應槽之材質、形成微 反應槽中之陣列塑微加熱器材料以及陣列型微加熱器之尺 寸係同前述之定義。此外，當可輕易理解的是，各種用以 導引樣品進入或離開晶片之反應槽之元件，例如微流道或 主動/被動閥門皆可視需求設置於晶片上。 以下實施態樣係用於進一步了解本發明之優點，並非 用於限制本發明之申讀專利範圍。 — 實施例：陣列型微加熱器之熱相分析 本實施例之陣列型微加熱器之材料與尺寸係如下：第 一金屬層使用鉑/鈦金屬，第二金屬層使用金/鈦金屬，晶片 之基材為玻璃（soda-lime glass)，第一層加熱器寬度為100 微求長度為6000微米，第二層加熱器長、寬度皆為100微 米。為達到晶片上的溫度自補償，最外側之第一層加熱器 寬度為200微米，第二層加熱器的長度為200微米。 偵測上述陣列型微加熱器之熱相為使用日本製之紅外 線熱相儀（型號：Infrared Thermography TVS-200N, Nippon Avinics Co Ltd.，Japan)進行量測而得，晶片上方覆蓋一層 100微米之玻璃片致使量測誤差降低並更能符合使用時之 實際/服度場。所得分析影像如第四圖所示。由圖中可知， 本發明之反應槽中所設置之陣列型微加熱器具有加熱均勻 200918897 之特性，對於其在_增幅3上的_能提升聚 反應之專一性與複製效率。 f. 綜合上述，本發明之反應槽中所設置之陣列型微加故 =具有良好賴均W，無須使科接系統進行溫度^ 償，經本設計即可直接進行溫度自補償。對於且 = 槽之生物晶片’特別是核酸增幅晶片，當可提反應 反應之專一性與複製致率。 升來δ轉鏈 甚他實施態樣 在本說明書中所揭露的所有特徵都可能鱼苴 二’本說明書中所揭露的每一個特徵都可“吉 同、相等或相似目的特徵所取代，因此， =的以相 特徵之外’翁的本說明書賴露的特徵 目^著的 特徵中的一個例子。 阳寻或相似 雖然本發明已以較佳實施例揭露如上，然其 限定本發明’任何熟悉此技藝者，在不脫離本發明，用以 和範圍内’當可作各種之更動與潤飾。 精神 10 200918897 【圖式簡單說明】 第一圖係為本發明之具溫度自補償功能之微反應槽所 具有之陣列型加熱器示意圖。 第二圖係為製備本發明之陣列型微加熱器之流程圖。 第三圖係為本發明之陣列型微加熱器達到熱補償之示 意圖。 第四圖係為本發明之陣列型加熱器之熱相圖。 [基本元件符號說明] 1 腔室表面 2 _ 第二·加熱層 2 ’ 第一加熱層之條狀微加熱器 3 第二加熱層 3 第二加熱層之陣列排列的微加熱器Self-compensation i, the micro-heater through the array design to achieve thermal uniformity and the four-three map, which is the top view of the first figure, the adjustment outside the picture and shame = due to the array design The temperature field of the partial region is consistent (the effect of heat equalization and temperature compensation, and then the temperature of the heating zone is increased, and the uniformity is improved). The biochip microreactor can be applied, for example, to a small sample to be heated, and has a sputum, +, and particularly nucleating nucleic acid amplifying wafer. Therefore, the present invention comprises a nucleic acid amplifying wafer which can be used as a microreactor. In addition to the array micro-heater of the present invention, the medium-two is easy to dissolve, and further includes a temperature 200918897 sensor for accurately measuring the reaction tank. The temperature is then used for temperature control purposes for polymerase chain reactions requiring precise temperature control. The material of the nucleic acid amplifying wafer of the present invention may be the same or different from the material of the microreactor provided thereon, the material of the microreactor, the array of micro-heater materials formed in the micro-reaction tank, and the array type micro-heater The dimensions are the same as defined above. In addition, it will be readily understood that various components, such as microchannels or active/passive valves, for guiding the sample into or out of the reaction cell of the wafer can be placed on the wafer as desired. The following embodiments are intended to further understand the advantages of the present invention and are not intended to limit the scope of the invention. - Example: Thermal phase analysis of the array type microheater The material and size of the array type microheater of the present embodiment are as follows: the first metal layer uses platinum/titanium metal, the second metal layer uses gold/titanium metal, and the wafer The substrate is made of soda-lime glass. The first layer heater has a width of 100 micrometers and a length of 6000 micrometers. The second layer heater has a length and a width of 100 micrometers. To achieve self-compensation of the temperature on the wafer, the outermost first layer heater has a width of 200 microns and the second layer heater has a length of 200 microns. The thermal phase of the array type micro-heater was detected by using an infrared thermal phase meter (Model: Infrared Thermography TVS-200N, Nippon Avinics Co Ltd., Japan) made in Japan, and the wafer was covered with a layer of 100 μm. The glass sheet reduces the measurement error and is more compatible with the actual/consistency field at the time of use. The resulting analysis image is shown in the fourth figure. As can be seen from the figure, the array type micro-heater provided in the reaction tank of the present invention has the characteristic of uniform heating of 200918897, and can enhance the specificity and replication efficiency of the polymerization reaction on the _amplification 3. f. In summary, the array type micro-addition provided in the reaction tank of the present invention has a good average W, and it is not necessary to make the temperature compensation of the joint system, and the temperature self-compensation can be directly performed by the design. For and = grooved biochips, especially nucleic acid amplification wafers, the specificity and replication rate of the reaction can be raised. All the features disclosed in this specification may be replaced by the characteristics of the same, the same or similar features. Therefore, An example of a feature that is characteristic of the specification of the present invention. The present invention has been disclosed in the preferred embodiment as above, but it defines the invention as any familiarity. The skilled person can use the invention to make various changes and refinements without departing from the scope of the invention. Spirit 10 200918897 [Simplified illustration] The first figure is a micro-reaction with temperature self-compensation function of the present invention. Schematic diagram of the array type heater provided in the tank. The second figure is a flow chart for preparing the array type microheater of the present invention. The third figure is a schematic diagram of the array type microheater of the present invention achieving thermal compensation. It is a thermal phase diagram of the array type heater of the present invention. [Basic component symbol description] 1 chamber surface 2 _ second · heating layer 2 ' strip heating micro heater 3 of the first heating layer 3 micro heaters arranged in an array of second heating layers