TWI809717B - Liquid cell microchip and measuring method thereof - Google Patents
Liquid cell microchip and measuring method thereof Download PDFInfo
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本發明係有關於半導體製程領域，尤其是關於一種適用於電子顯微鏡的液體量測載片的結構以及其量測方法。 The invention relates to the field of semiconductor manufacturing process, in particular to the structure of a liquid measuring slide suitable for an electron microscope and its measuring method.
利用穿透式電子顯微鏡(Transmission electron microscope，縮寫為TEM)或掃瞄電子顯微鏡(Scanning electron microscope，縮寫為SEM)等電子顯微鏡對材料層的掃描是常見的製程之一。上述電子掃描除了用於固體的材料層之外，也能用於掃描液態的液膜以用於觀察各種溶液狀態，例如半導體領域中可用於觀察研磨液或溶劑中的粒子大小、或是生物科技領域中可用於觀察各種生物液中的細胞狀態等，都是目前已經發展中的應用方式。 Scanning the material layer by using an electron microscope such as a transmission electron microscope (TEM for short) or a scanning electron microscope (SEM for short) is one of the common processes. In addition to the above-mentioned electronic scanning for solid material layers, it can also be used for scanning liquid liquid films to observe various solution states. In the field, it can be used to observe the state of cells in various biological fluids, etc., which are currently developing application methods.
由於電子顯微鏡的高真空和發射高能量的電子束會導致液體揮發，因此以電子顯微鏡觀察液體時，必須將液體填充至一液體量測載片中再進行觀察。其中液體量測載片由上下兩層載片基底組合成，填入待掃描的液體至兩載片基底之間的流道空間後，再將液體量測載片封裝並送至電子顯微鏡中進行掃描。因為液體位於密封的液體量測載片之中，因此不會產生液體揮發情況。 Since the high vacuum of the electron microscope and the emission of high-energy electron beams will cause the liquid to volatilize, when observing the liquid with the electron microscope, the liquid must be filled into a liquid measuring slide for observation. Among them, the liquid measurement slide is composed of upper and lower two slide substrates. After filling the liquid to be scanned into the flow channel space between the two slide substrates, the liquid measurement slide is packaged and sent to the electron microscope for scanning. scanning. Because the liquid is in the sealed liquid slide, there is no chance of liquid evaporation.
然而目前的技術中，使用電子顯微鏡掃描液體的製程技術仍有提升 的空間，例如提高掃描的解析度、簡化掃描步驟或是降低掃描的成本等。上述技術發展是本領域的研發目標之一。 However, in the current technology, the process technology of scanning the liquid with an electron microscope is still improving space, such as improving the scanning resolution, simplifying the scanning steps, or reducing the cost of scanning, etc. The above technical development is one of the research and development goals in this field.
因此，本發明提供一種改良的液體量測載片，可適用於各式電子顯微鏡的掃描步驟。本發明另外提供使用電子顯微鏡對液體量測載片進行掃描觀測的方法。 Therefore, the present invention provides an improved liquid measuring slide, which is applicable to the scanning steps of various electron microscopes. The present invention further provides a method for scanning and observing a liquid measuring slide using an electron microscope.
本發明提供一種液體量測載片，包含兩載片基底相互平行設置，其中兩載片基底之間包含有一流道空間，一液體灌入流道空間內，以及一第一電極以及一第二電極，分別設置於流道空間的兩個相對端點，並封住流道空間的兩個側邊。 The invention provides a liquid measuring slide, which comprises two slide substrates arranged parallel to each other, wherein a channel space is included between the two slide substrates, a liquid is poured into the flow channel space, and a first electrode and a second electrode , respectively set at two opposite end points of the flow channel space, and seal the two sides of the flow channel space.
本發明提供一種液體量測載片的量測方法，包含提供一液體量測載片，液體量測載片包含有兩載片基底相互平行設置，其中兩載片基底之間包含有一流道空間，一液體灌入流道空間內，一第一電極以及一第二電極，分別設置於流道空間的兩個相對端點，並封住流道空間的兩個側邊，以及提供一電子顯微鏡，且將液體量測載片送至電子顯微鏡內進行一量測步驟。 The invention provides a method for measuring a liquid measuring slide, which includes providing a liquid measuring slide, the liquid measuring slide includes two slide substrates arranged parallel to each other, and a flow channel space is included between the two slide substrates , a liquid is poured into the channel space, a first electrode and a second electrode are respectively arranged at two opposite ends of the channel space, and seal two sides of the channel space, and an electron microscope is provided, And the liquid measurement slide is sent to the electron microscope to perform a measurement step.
本發明的特徵在於，在液體量測載片的流道空間側壁形成導電層當作電極片，且液體量測載片的兩端使用導電膠封裝來當作電極接觸點。利用電子顯微鏡中聚能電子束本身帶電的特徵，在電子顯微鏡的腔體內部直接對液體量測載片進行通電。如此一來，可以在同一次電子顯微鏡的量測步驟中觀察到對液體量測載片未通電與通電的情況，達到節省成本與步驟的目的。此外本發 明還具有結構簡單容易製作的優點。 The present invention is characterized in that a conductive layer is formed on the side wall of the flow channel space of the liquid measurement slide as an electrode sheet, and both ends of the liquid measurement slide are packaged with conductive glue as electrode contact points. Utilizing the charged characteristics of the concentrated electron beam itself in the electron microscope, the liquid measurement slide is directly energized inside the cavity of the electron microscope. In this way, it is possible to observe in the same measurement step of the electron microscope that the liquid measurement slide is not energized and energized, thereby achieving the purpose of saving costs and steps. In addition this hair The invention also has the advantages of simple structure and easy fabrication.
1:液體量測載片 1: Liquid measuring slide
10:載片基底 10: slide substrate
12:凹槽 12: Groove
14:氮化矽層 14: Silicon nitride layer
16:電極層 16: Electrode layer
20:流道空間 20: runner space
21:流道空間開口 21: Runner space opening
22:液體 22: liquid
24:第一電極 24: The first electrode
26:第二電極 26: Second electrode
28:封口 28: seal
第1圖至第9圖繪示製作本發明的一液體量測載片的結構示意圖，其中：第1、3、6、7、9圖分別繪示不同製作階段液體量測載片的上視圖；第2圖繪示沿著第1圖中的剖面線A-A’所得的剖面圖；第4圖繪示沿著第3圖中的剖面線B-B’所得的剖面圖；第5圖繪示將兩個載片基底結合的剖面示意圖；以及第8圖繪示沿著第7圖中的剖面線C-C’所得的剖面圖。 Figures 1 to 9 show the structural schematic diagrams of making a liquid measuring slide of the present invention, wherein: Figures 1, 3, 6, 7, and 9 respectively show the top views of the liquid measuring slide at different production stages ; Fig. 2 shows a sectional view obtained along the section line A-A' in Fig. 1; Fig. 4 shows a sectional view obtained along the section line BB' in Fig. 3; Fig. 5 shows a schematic cross-sectional view of combining two slide substrates; and FIG. 8 shows a cross-sectional view taken along the section line CC' in FIG. 7 .
為使熟習本發明所屬技術領域之一般技藝者能更進一步了解本發明，下文特列舉本發明之較佳實施例，並配合所附圖式，詳細說明本發明的構成內容及所欲達成之功效。 In order to enable those who are familiar with the general skills in the technical field of the present invention to further understand the present invention, the preferred embodiments of the present invention are enumerated below, together with the accompanying drawings, the composition of the present invention and the desired effects are described in detail. .
為了方便說明，本發明之各圖式僅為示意以更容易了解本發明，其詳細的比例可依照設計的需求進行調整。在文中所描述對於圖形中相對元件之上下關係，在本領域之人皆應能理解其係指物件之相對位置而言，因此皆可以翻轉而呈現相同之構件，此皆應同屬本說明書所揭露之範圍，在此容先敘明。 For the convenience of description, the drawings of the present invention are only schematic diagrams for easier understanding of the present invention, and the detailed proportions thereof can be adjusted according to design requirements. As for the up-down relationship of relative elements in the figures described in the text, those skilled in the art should understand that they refer to the relative positions of objects, so they can be reversed to present the same components, which should all belong to this specification. The scope of disclosure is described here first.
第1圖至第9圖繪示製作本發明的一液體量測載片的結構示意圖，本發明所提供的液體量測載片，由兩個結構基本相同或類似的載片所結合而成。以下多數圖式中為了說明簡潔的目的，只繪出其中一個載片進行說明。首先請
參考第1圖以及第2圖，第1圖繪示液體量測載片的上視圖，而第2圖繪示沿著第1圖中的剖面線A-A’所得的剖面圖。提供一載片基底10，其中載片基底10的材質例如為矽，然後在載片基底10上以蝕刻等方式形成一凹槽12。之後在載片基底10上形成一氮化矽層14，也就是說氮化矽層14覆蓋於載片基底10的頂面、凹槽12的底面以及凹槽12的側壁。此處形成氮化矽層14的目的在於氮化矽層14可當作後續步驟中形成觀測窗的蝕刻停止層，關於這部分的技術內容屬於本領域的習知技術，在此不多贅述。
FIG. 1 to FIG. 9 are schematic diagrams showing the structure of a liquid measuring slide of the present invention. The liquid measuring slide provided by the present invention is formed by combining two slides with basically the same or similar structure. In most of the following figures, for the purpose of brevity, only one of the slides is drawn for illustration. first please
Referring to FIG. 1 and FIG. 2, FIG. 1 shows a top view of the liquid measuring slide, and FIG. 2 shows a cross-sectional view obtained along the section line A-A' in FIG. 1. A
從上視圖來看，本發明的凹槽12例如為十字形，此種凹槽形狀具有結構簡單的優點，然而本發明事實上並不限定凹槽12的形狀，也就是說可以依照實際需求調整凹槽12的形狀，各種不同形狀的凹槽也屬於本發明的涵蓋範圍。
From the top view, the
接著如第3-4圖所示，第3圖繪示液體量測載片的上視圖，第4圖繪示沿著第3圖中的剖面線B-B’所得的剖面圖。在載片基底10上例如以沉積等方式，全面性覆蓋一金屬材料層(圖未示)，然後對金屬材料層進行一回蝕刻步驟，以移除部分的金屬材料層，蝕刻後所留下的金屬材料層定義為電極層16。其中電極層16位於凹槽12內的氮化矽層14的側壁上。電極層16的材質例如為氮化鈦(TiN)、氮化鉭(TaN)、鎢(W)等導電材質，但本發明不限於此。
Next, as shown in Figures 3-4, Figure 3 shows a top view of the liquid measurement slide, and Figure 4 shows a cross-sectional view obtained along the section line B-B' in Figure 3. On the
到此步驟為止，已經形成包含有凹槽12、氮化矽層14以及電極層16的載片基底10，接著重複以上步驟形成另一個相似或是相同的結構，並且如第5圖所示，將兩個結構相同或是結構相似的載片基底10相互結合，其中兩個凹槽載片基底10的凹槽12相對設置，並且由凹槽12結合後所產生的一空間定義為流道空間20。本實施例中，流道空間20為一個十字形狀的密封空間，其中在後續
步驟中會將需要在電子顯微鏡下觀察的液體灌入流道空間20內後密封，以避免液體在電子顯微鏡的電子束之下迅速揮發，關於此特徵後續段落會進行進一步描述。另外本步驟中已經將兩個相似或相同的載片基底10相互結合，但後續圖式中為了簡潔說明，仍只繪出其中一個載片基底10，在此容先敘明。
Up to this step, the
如第6圖所示，為了要讓待觀察液體能注入流道空間20，將載片基底10的側邊進行切割並曝露出流道空間20，也就是說讓流道空間20產生開口，在此定義為流道空間開口21。本實施例中由於流道空間20為十字形狀，因此有四個流道空間開口21。接著如第7-8圖所示，第7圖繪示液體量測載片的上視圖，第8圖繪示沿著第7圖中的剖面線C-C’所得的剖面圖。將待觀察的液體22以針頭注射等方式注入流道空間開口21，由於毛細現象的關係，液體22會逐漸被吸入至流道空間20內部。
As shown in Figure 6, in order to allow the liquid to be observed to be injected into the
最後，如第9圖所示，將各流道空間開口21開口密封，以讓液體22固定在流道空間20內部避免液體揮發或是避免外界環境影響到液體22的狀態。本實施例中，將四個流道空間開口21分別密封，值得注意的是，其中兩個相對(也就是面對面且不相鄰)的流道空間開口21以導電膠密封，其中導電膠的材質例如為摻雜導電材料(例如銀)的矽膠或樹脂等，並且被導電膠密封後的部分定義為第一電極24以及第二電極26，而另外兩個剩下的流道空間開口21則以絕緣的封裝材料(材質例如為矽膠或是樹脂等)來密封，且密封後的流道空間開口21定義為封口28。後續再形成觀測窗等結構(這部分屬於習知技術不多加贅述)。到此步驟為止，已經完成本發明的液體量測載片1的基本結構。
Finally, as shown in FIG. 9 , the
請繼續參考第9圖，第一電極24與其相接觸的電極層16可以視為一電
極、而第二電極26與其相接觸的電極層16則可以視作另一電極，且在兩電極之間注入有待觀察的液體22。在實際應用上，可以將液體量測載片1送至電子顯微鏡(例如SEM或TEM，圖未示)進行觀察，該類電子顯微鏡均會發射帶電的電子束，若將電子束對準電極即可產生電壓，並對液體量測載片1進行通電。換句話說，本發明的特徵在於將電子顯微鏡所發射的電子束當作電壓源，對液體量測載片1進行通電。
Please continue to refer to Fig. 9, the
更詳細說明，在將液體量測載片1送至電子顯微鏡的腔體內之後，可以先進行一般的量測，也就是將電子束先對準流道空間20的中央位置，也就是說此時電子束並未對準電極(第一電極24或第二電極26)，故並不會產生電流流經液體量測載片1，可以先觀察液體22未通電前的狀態。接下來，在同一電子顯微鏡的同一腔體內，可以調整電子束的位置，例如將電子束對準第一電極24，而另外一個電極(第二電極26)在初始狀態下接地，就會產生一電流由第一電極24流向第二電極26，可以藉此觀察液體量測載片1被通電之後所產生的反應。舉例來說，在本發明的其中一種應用中可觀察液體22中的粒子的電性，若液體22具有帶電的粒子，則該些粒子會依據其帶有正電荷或是負電荷，而往不同的方向移動，若粒子不帶電則不會移動。在其他領域中，例如生物科技領域中也可能觀察細胞、細菌等微生物在通電之後的反應。
In more detail, after the
此外，上述步驟中先進行液體量測載片1未通電前的觀察，才進行通電後的液體量測載片1觀察。但本發明中也可以將順序對調，也就是先進行通電並觀察後，才進行未通電的觀察。這種步驟也屬於本發明的涵蓋範圍內。
In addition, in the above steps, the
綜合以上說明書與圖式，本發明提供一種液體量測載片1，包含兩載
片基底10相互平行設置，其中兩載片基底10之間包含有一流道空間20，一液體22灌入流道空間20內，以及一第一電極24以及一第二電極26，分別設置於流道空間20的兩個相對端點，並封住流道空間20的兩個側邊。
Based on the above instructions and drawings, the present invention provides a
在本發明的一些實施例中，其中液體量測載片用於一電子顯微鏡中，且電子顯微鏡包含有一電子槍，電子槍所發射的一電子束用於對第一電極24進行充電。
In some embodiments of the present invention, the liquid measuring slide is used in an electron microscope, and the electron microscope includes an electron gun, and an electron beam emitted by the electron gun is used to charge the
在本發明的一些實施例中，其其中從一上視圖來看，流道空間20為十字形狀。
In some embodiments of the present invention, the
在本發明的一些實施例中，其中第一電極24與第二電極26分別設置於十字形狀的流道空間20的相對的兩端點，且十字形狀的流道空間的另外兩個相對端點分別設置有絕緣封裝材料(即封口28)。
In some embodiments of the present invention, the
在本發明的一些實施例中，其中十字形狀的流道空間20的側壁設置有電極層16，且流道空間20的底部為氮化矽層14。
In some embodiments of the present invention, the sidewall of the
本發明另提供一種液體量測載片的量測方法，包含提供一液體量測載片1，液體量測載片1包含有兩載片基底10相互平行設置，其中兩載片基底之間包含有一流道空間20，一液體22灌入流道空間20內，一第一電極24以及一第二電極26，分別設置於流道空間20的兩個相對端點，並封住流道空間20的兩個側邊，以及提供一電子顯微鏡，且將液體量測載片送至電子顯微鏡內進行一量測步驟。
The present invention also provides a measurement method for a liquid measuring slide, which includes providing a
在本發明的一些實施例中，其中電子顯微鏡包含有一電子槍，電子槍所發射的一電子束用於對第一電極24進行充電。
In some embodiments of the present invention, the electron microscope includes an electron gun, and an electron beam emitted by the electron gun is used to charge the
在本發明的一些實施例中，其中將液體量測載片送至電子顯微鏡後，量測步驟包含以下步驟進行一第一階段量測，電子槍所發射的電子束對準流道空間20的一中央部分，並觀察液體量測載片1的情況，以及進行一第二階段量測，將電子槍所發射的電子束對準第一電極24，並觀察液體量測載片的情況。
In some embodiments of the present invention, after the liquid measurement slide is sent to the electron microscope, the measurement step includes the following steps to perform a first-stage measurement. The electron beam emitted by the electron gun is aimed at a part of the
在本發明的一些實施例中，其中第一階段量測與第二階段量測在電子顯微鏡的同一腔體內進行，且第一階段量測與第二階段量測之間並未進行一破真空步驟。 In some embodiments of the present invention, the first-stage measurement and the second-stage measurement are carried out in the same cavity of the electron microscope, and there is no vacuum break between the first-stage measurement and the second-stage measurement step.
在本發明的一些實施例中，其中更包含比對第一階段量測與第二階段量測時，電子顯微鏡所觀察到液體量測載片的變化。 In some embodiments of the present invention, it further includes the change of the liquid measurement slide observed by the electron microscope when comparing the first-stage measurement and the second-stage measurement.
本發明的特徵在於，在液體量測載片的流道空間側壁形成導電層當作電極片，且液體量測載片的兩端使用導電膠封裝來當作電極接觸點。利用電子顯微鏡中聚能電子束本身帶電的特徵，在電子顯微鏡的腔體內部直接對液體量測載片進行通電。如此一來，可以在同一次電子顯微鏡的量測步驟中觀察到對液體量測載片未通電與通電的情況，達到節省成本與步驟的目的。此外本發明還具有結構簡單容易製作的優點。 The present invention is characterized in that a conductive layer is formed on the side wall of the flow channel space of the liquid measurement slide as an electrode sheet, and both ends of the liquid measurement slide are packaged with conductive glue as electrode contact points. Utilizing the charged characteristics of the concentrated electron beam itself in the electron microscope, the liquid measurement slide is directly energized inside the cavity of the electron microscope. In this way, it is possible to observe in the same measurement step of the electron microscope that the liquid measurement slide is not energized and energized, thereby achieving the purpose of saving costs and steps. In addition, the present invention also has the advantages of simple structure and easy manufacture.
本發明的液體量測載片具有結構簡單的優點，除此之外藉由電子顯 微鏡本身所發射的電子束當作電壓來源以直接提供電流至液體量測載片，可以在電子顯微鏡的同一腔體內直接觀察同一個液體量測載片未通電前以及通電後的不同狀態。習知技術中，若要將液體量測載片進行通電觀察，則必須要外接一電源(例如電池)，且需要將液體量測載片從電子顯微鏡的腔體中移出，中間會先進行破真空、移出載片、重新抽真空、等待等步驟，如此一來既費時又會消耗更多成本，以實務面來看，以傳統方式欲進行載片的通電與未通電觀察比較實驗，至少需要兩次電子顯微鏡的掃描，無法在同一腔體內一次掃描步驟中完成，因此花費也是本發明的方法的兩倍。除此之外，將載片移出腔體後進行通電再重新送至腔體內的時間，載片也更容易受到外界環境的影響，因此有可能降低實驗的精確程度。總之，本發明具有結構簡單、省時、節省成本、高精確度等優點。 The liquid measuring slide of the present invention has the advantage of simple structure, in addition The electron beam emitted by the micromirror itself is used as a voltage source to directly provide current to the liquid measurement slide, and the different states of the same liquid measurement slide before and after power-on can be directly observed in the same cavity of the electron microscope. In the conventional technology, if the liquid measurement slide is to be energized and observed, an external power source (such as a battery) must be connected, and the liquid measurement slide needs to be removed from the cavity of the electron microscope, and the liquid measurement slide will be broken first. Steps such as vacuuming, removing the slide, re-vacuuming, and waiting are time-consuming and costly. From a practical point of view, if you want to conduct a comparative experiment between power-on and non-power-on observation of the slide in the traditional way, you need at least Two scans of the electron microscope cannot be completed in one scan step in the same cavity, so the cost is also twice that of the method of the present invention. In addition, the slide is more easily affected by the external environment after being powered on and then re-sent into the cavity after being removed from the cavity, which may reduce the accuracy of the experiment. In a word, the present invention has the advantages of simple structure, time saving, cost saving, high precision and the like.
以上所述僅為本發明之較佳實施例，凡依本發明申請專利範圍所做之均等變化與修飾，皆應屬本發明之涵蓋範圍。 The above descriptions are only preferred embodiments of the present invention, and all equivalent changes and modifications made according to the scope of the patent application of the present invention shall fall within the scope of the present invention.
1:液體量測載片 1: Liquid measuring slide
10:載片基底 10: slide substrate
16:電極層 16: Electrode layer
22:液體 22: liquid
24:第一電極 24: The first electrode
26:第二電極 26: Second electrode
28:封口 28: seal
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| TW201031911A (en) * | 2009-02-25 | 2010-09-01 | Nat Univ Chung Hsing | A capillary electrophoresis chip with integrated top-bottom opposed electrochemical sensing electrodes and its fabrication methods |
| CN110988048A (en) * | 2019-11-26 | 2020-04-10 | 江苏大学 | Cell activity evaluation device and method based on adhesion strength |
| US11249046B2 (en) * | 2014-01-17 | 2022-02-15 | The Board Of Trustees Of The University Of Alabama | Methods and systems for analysis |
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| TW201031911A (en) * | 2009-02-25 | 2010-09-01 | Nat Univ Chung Hsing | A capillary electrophoresis chip with integrated top-bottom opposed electrochemical sensing electrodes and its fabrication methods |
| US11249046B2 (en) * | 2014-01-17 | 2022-02-15 | The Board Of Trustees Of The University Of Alabama | Methods and systems for analysis |
| CN110988048A (en) * | 2019-11-26 | 2020-04-10 | 江苏大学 | Cell activity evaluation device and method based on adhesion strength |
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