200925271 八、發明說明： 【發明所屬之技術領域】 本發明係有_«種薄層_抹片製備暨原位雜交反應之裝 置’特別是有關於-用以製備薄層細胞抹片以及進行原位雜交反 應之薄層細胞抹片製備暨原位雜交反應之裝置。 Q 【先前技術】 子宮頸抹片是目前用於防治子宮頸癌之主要方法。傳統抹片 檢查疋用抹片刷刮取子宮頸細胞’塗於玻片上後送實驗室做病理 學判讀，雖然準確度不夠高’但幾乎所有的婦產科醫師都一致認 為，只要每年都去做，且連續三年都沒問題，一般而言，失誤的 機會其實是微乎其微。 ® 實施已有多年的傳統子宮職片檢查，其受賴界爭論的部 分疋其偽陰性高達UMG%，有部份是因為採樣技巧不夠好，另一 原因則疋受檢者本身的問題’例如子宮頸閉鎖、停經後子宮頸萎 ，敎未概剌蒙、子宮職讀屈或制，使得採取内子宮頸 細胞相當困難。 此外’如果抹>|上塗得太厚，亦即多層細胞重疊在—起，判 讀上也相當科’這—點是醫護人員必須加強再教育的。 另外一個常見的偽陰性原因是判讀失誤，由於一張抹片上有 200925271 义十萬、-田胞’要在短短幾分鐘之内做出判斷，實在不易。目 刖國内普遍是由細胞技 何貝職'人看片工作，先剔除掉他們認 -正常的片子，剩下-、兩成可能有酬的才交給醫師判讀。 ★近年來為了增加抹片神顧1學界_發料許多輔助 師檢方法’如薄層抹片ThinPrepTM及s_hTM等。薄層抹片是 職—奪子宮勒驗，特製保存财，歸_摔， ❹製成細胞分佈均勻的抹片再觀察。薄層抹片由於檢體細胞均句且 較不會有細胞重4現象，可增加峨病變的檢料，而文細胞集 中於較小區域内，對病理學判讀較為方便，可減少誤差，準確度 比傳統子宮頸抹片高，約為70-95%。 導致子宮職_子很多，最重要的域染人㈣突疲病毒 (HPV) ’幾乎所有的子宮頸癌都是HPV引起的。美國食物藥品 藝 e理局於2003年4月通過’年紀30以上的女性作子宮頸抹片筛 檢查時，可以選擇合併進行HPVDNA檢驗。也就是說，由子宮頸 採集的細胞除了檢查其外形與否異常外，也可以同時檢查是否有 高危險群的HPVDNA存在。 原位雜交法(ISH，in-situ hybridization)或螢光原位雜交法 (FISH，Fluorescencein-situ hybridization)，是將細胞固定在玻片上， 將玻片上細胞的細胞膜破壞，再應用探針(probe)於適當的條件 下’進入細胞内進行雜交。此探針可利用bi〇tin等標定或直接標定 200925271 榮光’以呈現不同顏色或螢光，再配合顯微鏡做最後賴。以原 位雜交技術進行HPV檢職不需要進行pcR放大，並可將細胞病 理染色結果與分子檢測結果加以整合’提供各個抹片細胞單獨之 檢測結果，增加臨床病理醫師判別及診斷上之進一步資訊，除可 檢測是否有病麵染外，可it—步得知jjpV縣基目是否嵌入細 胞染色質内，此為影響細胞病變之重要因素。 以傳統技術而言，如欲以薄層細胞抹片進行咖原位雜交法 檢測，必須先以™nprep™或SurePath™等所附裝置來製備薄層 細胞抹片樣品’織再將進行独雜交實驗所需之雜交框 (hybridization frame 或 hybridization chamber)等裝置附著於玻片 上’以進行雜交等後續反應^些操作步膽觀人力和物力成 本均較尚。 【發明内容】 鑒於上述背景之不便與缺失，為符合產業上某些利益之需 求，本發明&供一種薄層細胞抹片製備暨原位雜交反應之裝置， 可用以解決上述傳統之細胞抹片裝置未能達成之標的。 本發明之一目的在於提供一種薄層細胞抹月製備暨原位雜交 反應之裝置，藉由本發明所揭露之薄層細胞抹片製備暨原位雜交 200925271 反應之裝置可以簡化薄層細胞抹片製備及原位雜交實驗之操作步 驟，將其整合於同一裝置上。 根據以上所述之目的，本發明揭示了—種薄層細胞抹片製傷 暨原位雜交反狀裝置’薄層細胞抹#製備暨雜較反應之裝 置制以製備㈣細胞抹片以及進行原位雜交反應，其包含：一 々位裝置以及-密封裝置。定位裝置裝設於承載裝置上且定位裝 ❹ 置具有至少—翻，且孔洞之孔壁與承餘置形成—凹槽，其中， 凹槽係用以谷置-細轉浮液，藉由喃底部之交聯劑吸附細胞 懸浮液之細胞以形成—薄層細胞抹片。㈣裝置裝設於定位裝置 上用以密封凹槽以形成—封閉空間，封閉空間係用以進行原位雜 交反應。 ❹ 9 200925271 【實施方式】 根據以上所述之目的，本發明揭示了一種薄層細胞抹片製備 暨原位雜交反應之裝置，其制以製備薄層細麟#及進行原位 雜交反應之二合一實驗裝置。本發明在此所探討的方向為一種新 裝置’以簡化薄層細胞抹片製備及原位雜交實驗之操作步驟，將 其整σ於Π裝置上。為了能徹底地瞭解本發明，將在下列的描 ❹ 述巾提料盡的步驟及其組成。顯然地’本發明的施行並未限定 於該領域中具有通常知識者所熟習的特殊細節。另-方面，眾所 周知的組成或步驟並未描述於細節中，以避免造成本發明不必要 之限制。本發明的較佳實施例將會詳細描述如下，然而除了這些 詳細描述之外，本發明還可以廣泛地施行在其他的實施例中，且 本發明的範n不受限定，其以之後的專魏圍為準。 ❹ 如第- Α圖所示，本發明之第一實施例揭露一種薄層細胞 抹片製備暨原位雜交反應之裝置1〇〇，其包含：一定位裝置⑽以 及-密槌置13G。定位裝置12G具有至少—第—孔洞，當定位裝 置120裝設於承載裝置110上時，第一孔洞之孔壁與承載裝置110 形成凹槽，承載裝置11〇塗佈一交聯劑，本實施例中較佳範例 承載裝置110為玻片，交聯劑選自下列之-者或其組合：左旋離 胺酸（P〇ly-L-lysine)或魏（silane)，其中，凹槽係用以容置一 、田胞懸雜’藉細槽底狀交麵韻細_雜之細胞以形 200925271 成薄層細胞抹片。密封裝置130裝設於定位裝置120上用以密 封凹槽以域封閉空間，且於封閉空間進行原位雜交反應。 如第B圖及第-C _示，上述之定位裝置12G具有五 層、、口構五層結構依次由上到下分別為：第—離型紙層⑵A、第 黏膠層124A、第—基材泡棉層126、第二黏膠層12犯以及第二 離▲層122B。其巾，上述之第二離型紙層用以保護第二 ❹黏膠層124B胃將第二離型紙層122B撕下後，第二黏膠層124B 與承載裝置110黏合。此外，第一離型紙層隐用以保護第一黏 膠層124A’田將第一離型紙層122A撕下後，第一黏膠層^與 密封裝置130黏合。上述之第一基材泡棉層厚度約為〇〇5 ·〜5 mm ’其選用的材料為下列之一者或其組合：聚氨§旨⑽；_ urethane)、聚氣乙烯（PVC ; p〇lyVinyl Chl〇ride)、聚乙烯（pE ; polyethylene )、聚乙烯對苯二甲酸酯（pET ; ρ〇ι^%ι_ 0 Terephthalate)以及NeopreneAcrylic。上述之第一黏膠層以及第二 黏膠層為壓克力膠。 如第二A圖、第二B圖及第二C圖所示，為本實施例之另 一範例，其中，上述之薄層細胞抹片製備暨原位雜交反應之裝置 1〇〇更具有-增厚裝置200，其中’增厚裝置2〇〇裝設於定位裝置 120上且具有至少一第二孔洞’當增厚裝置2〇〇與定位裝置12〇結 合時，第一孔洞與第二孔洞互相疊合。上述之增厚裝置2〇〇具有 200925271 兩層結構分別為一第二基材泡棉層2i〇以及一第三黏膠層22〇,第 二黏膠層220與第一離型紙層122A黏合。其中，第二基材泡棉層 厚度約為0.05 mm〜5 mm，且材料為選自下列之一者或其組合： 聚氨醋（PU; poly urethane )、聚氣乙烯（PVC; P〇ly Vinyl Chloride )、 聚乙烯(PE;polyethylene)、聚乙烯對苯二甲酸酯（PET;p〇1yethylene200925271 VIII. Description of the invention: [Technical field to which the invention pertains] The present invention relates to a device for preparing a thin layer smear preparation and in situ hybridization reaction, in particular, for preparing a thin layer of cell smears and performing the original A device for preparing a thin layer of cell smears and in situ hybridization reaction. Q [Prior Art] Pap smear is currently the main method for the prevention and treatment of cervical cancer. Traditional smear examination, smear the cervical cells with a smear brush, and then sent to the laboratory for pathological interpretation, although the accuracy is not high enough, but almost all gynecologists agree that as long as every year Do, and for three consecutive years no problem, in general, the chance of mistakes is actually very small. ® has implemented traditional uterine film examinations for many years, and its part of the dispute has been falsely negative as high as UMG%, partly because sampling techniques are not good enough, and another reason is the problem of the subject itself' Cervical atresia, cervix dystrophy after menopause, sputum sputum, uterine job reading or system, making it difficult to take the inner cervical cells. In addition, if the wipe is too thick, that is, the multi-layer cells overlap, the interpretation is quite scientific. This point is that medical personnel must strengthen re-education. Another common false negative cause is the interpretation error. It is not easy to make judgments in just a few minutes on 200925271. In the domestic market, it is generally the case that the cell technology is used to read the film. The first thing is to remove the normal film, and the rest, and the two may be paid to the doctor. ★ In recent years, in order to increase the smear of the gods 1 academic _ a lot of auxiliary inspection methods such as thin layer smear ThinPrepTM and s_hTM. The thin layer of smear is the job - the uterus is checked, the special saves the wealth, and the _ falls, and the sputum is made into a uniform distribution of cells and then observed. Thin layer smear can increase the detection of sputum lesions due to the fact that the sample cells are uniform and have no cell weight 4 phenomenon, while the corpuscles are concentrated in a small area, which is convenient for pathological interpretation, which can reduce errors and accuracy. The degree is higher than the traditional Pap smear, about 70-95%. Leading to the uterus, a lot of people, the most important domain is infected with humans. (4) Acute virus (HPV) ‘Almost all cervical cancers are caused by HPV. The United States Food and Drug Administration e. In April 2003, when the women who were older than 30 years were screened for Pap smear, they could choose to combine HPV DNA tests. That is to say, in addition to checking whether the shape of the cells collected by the cervix is abnormal, it is also possible to check whether there is a high risk group of HPV DNA. In situ hybridization (ISH) or Fluorescence in-situ hybridization (FISH) is to immobilize cells on a slide, destroy the cell membrane of the cells on the slide, and then apply the probe (probe). Under the appropriate conditions, 'into the cells for hybridization. This probe can be calibrated with bi〇tin or directly calibrated 200925271 glory to present different colors or fluorescent light, and then cooperate with the microscope to make the final decision. In situ hybridization for HPV examination does not require pcR amplification, and can integrate cytopathic staining results with molecular detection results to provide separate detection results for each smear cell, and to increase further information on clinician discriminator diagnosis and diagnosis. In addition to detecting whether there is a disease surface, it can be found that the jjpV county base is embedded in the cell chromatin, which is an important factor affecting cell pathology. In traditional technology, if you want to use a thin-layer cell smear for in-situ hybridization detection, you must first prepare a thin-layer cell smear sample with a device such as TMnprepTM or SurePathTM. The device such as the hybridization frame or the hybridization chamber required for the experiment is attached to the slide glass to perform subsequent reactions such as hybridization, and the human and material costs are relatively high. SUMMARY OF THE INVENTION In view of the inconvenience and lack of the above background, in order to meet the needs of certain interests in the industry, the present invention provides a device for preparing a thin layer of cell smears and in situ hybridization reaction, which can be used to solve the above conventional cell wipes. The device failed to achieve the target. An object of the present invention is to provide a device for thin layer cell smear preparation and in situ hybridization reaction, which can simplify the preparation of thin layer cell smears by the thin layer cell smear preparation and in situ hybridization 200925271 reaction device disclosed in the present invention. And the steps of the in situ hybridization experiment, integrated into the same device. According to the above object, the present invention discloses a thin layer cell smear wounding and in situ hybridization device "thin layer cell smear # preparation and heterogeneous reaction device to prepare (4) cell smear and original A bit hybridization reaction comprising: a clamping device and a sealing device. The positioning device is mounted on the carrying device and the positioning device has at least a turn-over, and the hole wall and the bearing hole are formed into a groove, wherein the groove is used for the valley-fine transfer floating liquid, The crosslinker at the bottom adsorbs the cells of the cell suspension to form a thin layer of cell smears. (4) The device is mounted on the positioning device for sealing the groove to form a closed space for the in-situ hybrid reaction. ❹ 9 200925271 [Embodiment] According to the above object, the present invention discloses a device for preparing a thin layer of cell smears and an in situ hybridization reaction, which is prepared by preparing a thin layer of fine lining # and performing in situ hybridization reaction. Integral experimental device. The present invention is directed to a novel device' to simplify the steps of thin cell smear preparation and in situ hybridization experiments, which are sigmaized on a sputum device. In order to thoroughly understand the present invention, the steps and compositions of the towels will be described in the following. It is apparent that the practice of the present invention is not limited to the specific details that are common to those skilled in the art. In other instances, well-known components or steps are not described in detail to avoid unnecessarily limiting the invention. The preferred embodiments of the present invention will be described in detail below, but the present invention may be widely practiced in other embodiments, and the scope of the present invention is not limited. Wei Wei is the standard. As shown in the first section, a first embodiment of the present invention discloses a thin layer cell smear preparation and in situ hybridization reaction apparatus comprising: a positioning device (10) and a densely packed 13G. The positioning device 12G has at least a first hole. When the positioning device 120 is mounted on the carrier device 110, the hole wall of the first hole forms a groove with the carrier device 110, and the carrier device 11 is coated with a crosslinking agent. The preferred example carrier device 110 is a slide, and the crosslinking agent is selected from the group consisting of: P〇ly-L-lysine or silane, wherein the groove is used. In order to accommodate one, the field cell suspension is a small cell smear of the shape of 200925271. The sealing device 130 is mounted on the positioning device 120 for sealing the recess to close the space and perform an in situ hybridization reaction in the enclosed space. As shown in FIG. B and FIG. 3C, the positioning device 12G has five layers and five layers of mouth structure, which are sequentially from top to bottom: a first release paper layer (2) A, a second adhesive layer 124A, and a first base. The material foam layer 126, the second adhesive layer 12 and the second ▲ layer 122B. The second release layer is used to protect the second release layer 124B. After the second release layer 122B is torn off, the second adhesive layer 124B is bonded to the carrier 110. In addition, the first release layer is used to protect the first adhesive layer 124A'. After the first release liner 122A is torn off, the first adhesive layer is bonded to the sealing device 130. The first substrate foam layer has a thickness of about ·5·~5 mm. The material selected is one of the following or a combination thereof: polyamine § (10); _ urethane), polyethylene gas (PVC; p 〇 lyVinyl Chl〇ride), polyethylene (pE; polyethylene), polyethylene terephthalate (pET; ρ〇ι^%ι_ 0 Terephthalate) and Neoprene Acrylic. The first adhesive layer and the second adhesive layer are acrylic adhesive. As shown in FIG. 2A, FIG. 2B and FIG. 2C, another example of the present embodiment is that the apparatus for preparing a thin layer of cell smears and in situ hybridization has the same- The thickening device 200, wherein the 'thickening device 2 is mounted on the positioning device 120 and has at least one second hole'. When the thickening device 2 is combined with the positioning device 12, the first hole and the second hole Overlapping each other. The thickening device 2 has the 200925271 two-layer structure of a second substrate foam layer 2i and a third adhesive layer 22, and the second adhesive layer 220 is bonded to the first release layer 122A. Wherein, the second substrate foam layer has a thickness of about 0.05 mm to 5 mm, and the material is one or a combination selected from the group consisting of: polyurethane (polyurethane) (polyurethane), polystyrene (PVC); Vinyl Chloride ), polyethylene (PE; polyethylene), polyethylene terephthalate (PET; p〇1yethylene)
Terephthalate )以及 NeopreneAcrylic 〇 上述薄層細胞抹片製備暨原位雜交反應之裝置，上述裝置有 二種型式，第一種型式是只具有單層定位裝置，其定位裝置與密 封裝置結合於一承載裝置上以製備薄層細胞抹片並進行原位雜交 反應。第二種型式是具有複數層定位裝置，藉由複數層定位裝置 以增加定位裝置内部容納懸浮細胞的空間。第三種型式是定位裝 置更具有一增厚裝置，藉由增厚裝置可以達到與第二型式的目的 相同，增厚裝置的厚度可以任意調整。上述之複數層定位裝置以 及增厚裝置的设置方法有以下兩種：第一種，定位裝置可以在未 與承載裝置結合前，藉由複數層定位裝置或另加一增厚裝置來增 厚。第二種，定位裝置與承載裝置結合後，再與另外之定位裝置 或增厚裝置結合以達增厚目的。 本發明揭露一種薄層細胞抹片製備暨原位雜交反應之裝 置，其中以薄層細胞抹片製備暨原位雜交反應之裝置製備薄層細 胞抹片以及進行原位雜交反應之方法包含下列：首先，提供一承 12 200925271 載裝置110，接著進行一塗佈程序，塗佈交聯劑於承載裝置之一表 面，待交聯劑乾燥後，將定位裝置120裝設於承載裝置上以形成 凹槽。 於塗佈程序完成後’將置於保存财之採樣細胞製備成一細 胞懸洋液，並將細胞懸浮液加入凹槽。接著，進行一靜置程序， #由交聯劑爾細胞懸浮液中沈降之採樣細胞以形成薄層細胞抹 ❹ 灸進行雜交程序，藉由添加探針並以密封裝置130裝 »又於疋位裝置上，鄉成—賴朗而進行原姉交反應。 如第三A圖、第三B圖以及第三c圖所示，為本實施例之 再較佳範例，疋位裝置裝具有至少一孔洞或數個孔洞，不同的 孔洞内可以進行不同的檢測。其中，孔洞其形狀為下列之-者： 圓形方形—肖形、楔形或任意幾何形狀。但本發明並不以此 為限。 ❿ 範例一 •丨.將玻片（玻璃或其他聚合材質之娜玻片）塗覆/被覆（⑶ating) 父聯劑’其目的係用㈣細胞軸其上。將玻片浸潤在 P y lysinei^ia^e等溶液中1〇分鐘，取出後置於室溫下暗乾。 2.將置於保存液翻定液之採樣細胞離心沈降，㈣細胞後，重新 懸浮於緩衝液中，以製備成細麵浮液（Cell suspension)。 13 200925271 3·將定位裝置之離型紙剝離並與玻片黏合，加入細胞懸浮液，靜置 15分鐘使細胞自然沈降吸附於玻片上，然後倒掉細胞懸浮液，以 緩衝液清洗一次後，置於室溫下風乾。Terephthalate) and Neoprene Acrylic 装置 the above-mentioned thin-layer cell smear preparation and in situ hybridization reaction device. The above device has two types, the first type has only a single layer positioning device, and the positioning device and the sealing device are combined with a bearing device. The thin layer of cell smears were prepared and subjected to an in situ hybridization reaction. The second type has a plurality of layer positioning means for increasing the space for accommodating suspended cells inside the positioning means by means of a plurality of layer positioning means. The third type is that the positioning device has a thickening device which can be achieved by the thickening device in the same manner as the second type, and the thickness of the thickening device can be arbitrarily adjusted. The above-mentioned plurality of layer positioning devices and the thickening device are arranged in the following two ways: First, the positioning device can be thickened by a plurality of layer positioning devices or an additional thickening device before being combined with the carrier device. Secondly, after the positioning device is combined with the carrying device, it is combined with another positioning device or thickening device for the purpose of thickening. The invention discloses a device for preparing a thin layer cell smear and an in situ hybridization reaction, wherein the method for preparing a thin layer cell smear by using a thin layer cell smear preparation and an in situ hybridization reaction device and performing the in situ hybridization reaction comprises the following: First, a 12 200925271 carrier device 110 is provided, followed by a coating process, applying a crosslinking agent to one surface of the carrier device, and after the crosslinking agent is dried, the positioning device 120 is mounted on the carrier device to form a concave surface. groove. After the coating procedure is completed, the sampled cells, which are placed in the preserved state, are prepared into a cell suspension and the cell suspension is added to the groove. Next, a rest process is performed, # sampling cells settled in the cross-linker cell suspension to form a thin layer cell smear moxibustion hybridization procedure, by adding a probe and sealing the device 130 On the device, the township-Lailang conducts the original cross-linking reaction. As shown in FIG. 3A, FIG. 3B, and FIG. 3C, in a further preferred embodiment of the present embodiment, the clamping device is provided with at least one hole or a plurality of holes, and different holes can be detected differently. . Among them, the shape of the hole is as follows: Round square - Xiao, wedge or any geometric shape. However, the invention is not limited thereto.范例 Example 1 • 丨. Apply or cover a slide (glass or other polymeric material) (the (3) ating) parental agent's purpose is to use (4) the cell axis. The slide was soaked in a solution such as P y lysinei ^ ia ^ e for 1 minute, taken out, and left to dry at room temperature. 2. Centrifuge the sampled cells placed in the preservation solution, and (4) the cells, and then resuspend in the buffer to prepare a Cell suspension. 13 200925271 3. Remove the release paper from the positioning device and bond it to the slide. Add the cell suspension and let stand for 15 minutes to allow the cells to naturally settle and adsorb on the slide. Then pour off the cell suspension and wash it once with buffer. Dry at room temperature.
4.加入含有螢光標定核酸探針之雜交緩衝液，剝離離型紙後以密封 裝置密封’於37。(：進行雜交反應，4小時後將此定位及密封裝置 組合自玻片上移除，以0·2χ SSC緩衝液於55〇C下清洗，再經MQ 清洗後置於室溫下晾乾’以螢光顯微鏡觀察結果。 範例二 1. 將玻片（玻璃或其他聚合材質之塑膠玻片）塗覆/被覆（c〇ating) 交聯劑，其目的係用以將細胞黏附其上。將玻片浸潤在 poly-L-lysme或silane等溶液中1〇分鐘，取出後置於室溫下晾 乾’再與定位裝置組合。 2. 將置於保存液或固定液之採樣細胞離心沈降，收集細胞後，重 新懸浮於緩衝液中’以製備成細胞懸浮液（CeUsuspensi〇n)。 3·將具有增厚裝置之定位裝置之_紙_並與玻片黏合，加入 細胞懸浮液，靜置15分鐘使峨自然沈降吸_玻片上，然 後倒掉細麟浮液，錢衝液清洗—錢，置於室溫下風乾。 4.移除增厚裝置以及定位裝置之_紙，加人含有螢光標定核酸 探針之雜級衝液，以密封裝置密封於3η：進行誠反應，4 200925271 小時後將此定位及密封裝置組合自玻片上移除，以〇.2χ SSC緩 衝液於55°C下清洗，再經MQ清洗後置於室溫下晾乾，以榮光 顯微鏡觀察結果。 本發明可能有許多的修 顯然地，依照上面實施例中的描述，4. Add the hybridization buffer containing the fluorescent probe to the nucleic acid probe, peel off the release paper, and seal with a sealing device at 37. (: The hybridization reaction was carried out. After 4 hours, the positioning and sealing device combination was removed from the slide, washed with 0. 2 χ SSC buffer at 55 ° C, then washed with MQ and then allowed to dry at room temperature. Fluorescence microscopy observations. Example 2 1. Coating/coating a glass slide (glass or other polymeric plastic glass slide) with a cross-linking agent for the purpose of adhering cells to it. The tablets are infiltrated in a solution such as poly-L-lysme or silane for 1 minute, taken out and left to dry at room temperature, and then combined with a positioning device. 2. Centrifuge and collect the sampled cells placed in the preservation solution or fixative solution. After the cells are resuspended in the buffer to prepare a cell suspension (CeUsuspensi〇n). 3. Place the paper with the device of the thickening device and bond it to the slide, add the cell suspension, and let stand 15 Minutes make the cockroaches naturally settle on the _ slide, then pour off the fine lining float, clean the money with money - money, and let it air dry at room temperature. 4. Remove the thickening device and the positioning device _ paper, add people with fluorescent Calibration of the heterogeneous flush of the nucleic acid probe, sealed to 3η with a sealing device: After the honest reaction, 4 200925271 hours, the positioning and sealing device combination was removed from the slide, washed with 〇.2χ SSC buffer at 55 ° C, then washed with MQ and then allowed to dry at room temperature to glory Microscopic observations. The invention may have many modifications, as described in the above embodiments,
Φ 正與差異。因此需要在其附加的權利要求項之範圍内加以理解， 除了上述詳細的描述外，本發明還可以廣泛地在其他的實施例中 施打。上述僅為本發明之較佳實關而已，並_嫌林發明 之申請專利制；凡其它未麟本發騎揭示之卿下所完成的 等效改變絲飾，均應包含在下射請專纖圍内。 【圖式簡單說明】 第—A圖係本發明之薄層細胞抹片 裝置及承錄置之外齡意圖； -觀敎反應之 裝置之I:，本發明之薄層細胞， 裝置及製備暨原位雜交反應之 戳展置之組合側視圖； 係本發明之增厚裝置及薄層細 第二A圖 胞祙片製備暨原位 圖 雜交反應之裝置之侧視 200925271 第4圖係本發明之增厚裝置及薄層細跑抹片製備暨原位 雜交反應之裝置組合侧視圖； 、 第二c圖係本發明之薄層細胞抹片製備暨原位雜交反應之 裝置及承載裝置之組合侧視圖； f 第三A圖係本發明之具有一孔洞之定位裝置； ’ 圖係本發明之具有二孔洞之定位裝置；以及 • $三C ®縣_之具有三孔敗定位裝置。 【主要元件符號說明】 2層細胞抹片製備__交反應之裝置 了 B- 100 110 120定位裝置 122A第一離型紙層 122B第二離型紙層 124A第一黏膠層 Φ 124B第二黏膠層 126第一基材泡棉層 130密封裝置 200增厚裝置 * 210第二基材泡棉層 220第三黏膠層Φ Positive and negative. It is therefore to be understood that within the scope of the appended claims, the invention may be The above is only a better implementation of the present invention, and the patent application system of the invention of the invention; the equivalent change silk decoration completed by the other uncles of the lining of the lining of the lining should be included in the lower beam. Inside. BRIEF DESCRIPTION OF THE DRAWINGS The first-A diagram is a thin-layer cell smear device of the present invention and the intention of recording the external age; - the device for the observation reaction I: the thin-layer cell, device and preparation of the present invention Side view of a combination of in-situ hybridization reaction; a side view of a thickening device of the present invention and a thin layer of a second A-tube preparation and in situ hybridization reaction 200925271 FIG. 4 is a view of the present invention Side view of the device for thickening device and thin layer fine running smear preparation and in situ hybridization reaction; and second c picture is a combination of the device for preparing thin layer cell smears and in situ hybridization reaction device and carrier device of the invention Side view; f Third A is a positioning device having a hole in the present invention; 'Figure is a two-hole positioning device of the present invention; and • Three C® County has a three-hole failure positioning device. [Main component symbol description] 2 layer cell smear preparation __ AC reaction device B-100 110 120 positioning device 122A first release paper layer 122B second release paper layer 124A first adhesive layer Φ 124B second adhesive Layer 126 first substrate foam layer 130 sealing device 200 thickening device * 210 second substrate foam layer 220 third adhesive layer