1292822 九、發明說明： 【發明所屬之技術領域】 更特別地係為了質譜分 本發明係關於—種質譜技術 析而產生氣相帶電荷分子。 【先前技術】 質譜技術可根據其質荷比（常以崎或▲表示）來 ，定分子，因此在進行f譜分析時需探討一電場中的帶電 荷分子性質。帶電荷分子性質可由其質荷比判斷。例如在 四極離子阱（quadrupole ion-trap)質譜儀中，帶電荷分子 被離子阱捕捉，一施加於被捕捉分子之外加電場造成分子 依一方式運作，藉此指出其質荷比（以w/名或表示）。 藉由偵測被捕捉的分子之質荷比，分子的質量可被檢測， 由此可鑑定該分子。 為產生質譜分析之帶電荷分子，常見於質譜將分子離 子化之技術已經被研究，且提供其中至少部分氣相形式之 帶電荷分子。離子化並產生氣相分子之技術已經被探討， 例如基質輔助雷射脫附離子化（Matrix Assisted Laser Desorption Ionization ’ MALDI )，其可能造成樣品中的基質 分子被導入氣相分子中。此外，離子化過程經常產生目標 分子的碎裂及部分的破壞。又對分子之樣品進行一離子化 程序經常增加質譜圖像的複雜度。 【發明内容】 一方面，本發明之製造氣相分子方法包含提供一分子 5 1292822 之樣品，該樣品係具有獨特之電荷分佈，及導引聲波輻射 至該分子之樣品，以由該樣品中脫附至少部分分子，如此 該被脫附分子具有一與分子樣品中實質上相同之電荷分 佈。 實施態樣中可包含一個或多個下列態樣： 導引聲波輻射包含提供能量至一基材以誘發基材中之 聲波。 樣品係設置於基材上。在部分實施態樣中，基材包含 石夕。 樣品係設置於基材之一表面，且能量係由基材之另一 面提供。 提供基材能量包含產生電磁輻射及導引該電磁輻射至 基材之一表面以誘發基材内聲波。電磁輻射可包含雷射輻 射。 提供能量包含產生一電子束及導引該電子束至基材之 一表面以誘發基材内聲波。 提供能量包含造成基材一表面之機械式震盪以誘發基 材内聲波。例如，一壓電元件可用來造成機械式震盪。 導引聲波輻射包含利用例如連續音波源、超音波源及/ 或脈衝源產生聲波。 導引聲波輻射之操作係與導引離子化輻射至樣品並非 同時。 在部分實施態樣中，該方法進一步包含對被脫附分子 操作質譜分析，質譜分析可包含飛時質譜分析、四極質譜 分析、離子阱質譜分析、磁扇形場式質譜分析、傅立葉轉 1292822 換離子迴旋加速共振質譜分析及/或離子泳動質譜分析。 在另一方面，本發明之製造氣相分子裝置，其係包含 一容納一分子樣品之容器（receptacle)，該樣品係具有獨特之 電荷分佈；及一聲波源，其係設置用以導引聲波輻射至分 子之樣品，以造成樣品中至少部分分子被脫附。被脫附的 分子具有一實質上與前述分子之樣品中相同之電荷分佈。 又一方面，本發明之製造氣相分子方法，其係包含提 供一分子之樣品，利用一連續音波源、一超音波源及/或一 脈衝源產生聲波輻射；及導引前述聲波輻射至前述分子之 樣品以脫附樣品中之至少部分分子。聲波輻射係射入至分 子之樣品。該方法也包含離子化前述分子。 再一方面，本發明之製造氣相分子裝置，其係包含一 容器用以容納一分子之樣品；及一聲波輻射產生器，該聲 波輻射產生器係設置用以導引聲波輻射至前述分子之樣 品，以造成至少部分分子由樣品中脫附。聲波輻射產生器 包含例如一連續音波源、一超音波源及/或一脈衝源。 再一方面，本發明之製造氣相分子方法，其係包含提 供一分子之樣品；及導引聲波輻射至前述分子之樣品，以 在無前述樣品離子化程序下由樣品中脫附至少部分分子。 至少部分被脫附之分子具有一電荷。被脫附的分子係進行 質譜分析。 再一方面，本發明之製造氣相分子裝置，其係包含一 容器以容納一分子之樣品；及一聲波源，該聲波源係設置 用以導引聲波輻射至前述分子之樣品，以造成至少部分分 子由樣品中脫附。至少部分被脫附的分子具有一電荷。一 7 1292822 ^譜儀係可被設置使用來分析前述被脫附之分子。前述樣 品係未遭受一離子化程序下暴露於聲波輻射。 本發明之-個或多個實施態樣之細節係於圖式與以下 ^述中被提出。本發明之其他特徵、目的及優點係由以下 敌述、圖式及申請專利範圍中顯而易見。 【實施方式】 第一圖係為一利用質譜儀分析使用聲波輻射脫附（即 由提供的樣品中釋放）之分子之範例裝置1〇〇之前視圖。 第二圖係為第一圖裝置之區塊概要圖（基於簡化圖式 的目的，部分第一圖中的零件並未顯示於第二圖中）。如 第—圖與第二圖所示，裝置10〇包含一聲波源109，其係導 引聲波能量至一分子之樣品忉5以脫附樣品中至少部分之 分子。被脫附之分子因此形成氣相分子而被裝置1〇〇的質 譜儀量測與偵測設備119分析。可輕易理解地，至少部分 被脫附的分子具有一電荷，更進一步，在部分實施態樣中， 被脫附的分子的電荷分佈實質上與樣品105中的電荷分佈 相同。 為製造氣相分子’需有一容器容納分子之樣品105。分 子之樣品105係設置於一接近質譜儀設備119,使得部分分 子能於被釋放後導引入質譜儀量測與偵測設備U9中。在 第一圖中，樣品係設置一四極離子阱102 (Quadmpole Ion Trap ( QIT)之管道或導管之開口端（圖未顯示）’藉此導 入qIT 1〇2的内部區域103，四極離子阱1〇2之内部區域103 係為質譜分析操作之處。 8 1292822 由聲波源1G9製造之聲波輻射接著導引至分子之樣品 105上。部分樣品105中的分子隨之獲得足夠動能使其能由 樣品105的主體中被脫附或釋放。那些被脫附的分子係被 導引進入QIT 102之内部1〇3區域。 利用聲波輕射製造質譜分析之氣相分子一般無需將樣 品離子化。此個為任何減供的分子#品將有至少部分 分子已經離子化’ a此具有一正電荷或負電荷（此類不必 經由離子化程序即具有電荷之分子有時被稱為“天 ^(b_-charge)分子。事實上也有樣品之化性被視為無 刀子之W生才取口口乃至於—離子化樣品中的高 八 子，其令的電荷分子濃度變化極矩，然在所有樣口:二 有部分分子係帶有電荷。因此’任何被提供之分^ ^ 具有一獨特之電荷分佈。 刀于之枚口口 當聲波被施加於一分子之樣品時，聲八 面的鍵結，因此造成分子由樣品⑽中被釋放=== 下特別討論的雷射誘發聲波脫附中，誘發波㈣ ^盘 結振動頻率被聲波誘發的範圍近似，因 此，雷㈣料波可有效破壞分子表⑽結。此外，^ 誘發聲波頻率與表面鍵結振動頻率間的一致性可 ^射 能量的傳遞，ϋ此避免能量轉換的損失或失效，這= 生在製造氣相分子的常見技術中。更進一步，聲波^吊， 在無分手砰裂下將分子平穩脫附。在部分實施能樣寸I =的被脫附分子具一實質上與樣品中分子相同“的電荷分 9 1292822 儘管使用聲波脫附而不伴隨任何其他離子化可能僅製 造出少量的氣相分子，然而基於質譜分析的目的時，即使 少量的帶電荷形成氣相即足夠，因為質譜儀裝置只需要少 量的帶電荷分子即足以鑑定這些分子相對的w々值。儘管帶 電荷離子相對於中性分子/粒子的比例低，但在一些例子 中，其整體偵測效率與利用離子化裝置的質譜儀相比近 似，甚至更佳。在利用離子化裝置的質譜儀進行生物分子 或生物粒子的質譜分析令，其離子化率一般來說是很低 的。例如當使用MALDI或電喷灑離子化（ESI)進行生物 分子的離子化效率一般低於0·0001。對於部分大生物分子， 效率可能接近0。例如，大聚醣分子（M> 100,000 Daltons) 使用一般離子化裝置無法適切地離子化。 更進一步，儘管聲波脫附造成不帶電荷分子也被釋放 到氣相煙霧中，但氣相中不帶電荷分子的存在並不會影響 質譜分析結果，因為不帶電荷的分子並不會被電荷粒子偵 測器偵測到，此外，不帶電荷分子並不會被離子阱裝置捕 捉。 在此所描述的聲波脫附可應用於任何類型的分子，包 含： 1) 在質譜儀操作溫度下不具有足夠氣壓之液態樣品與 無定形材料，這些液態樣品與不定形材料包含液晶與 離子液體（ionic liquids)； 2) 被固體狀態樣品捕捉的分析物，例如被捕捉於金屬箔 片中的氮， 3) 基材表面的被吸附物，例如催化金屬表面的自由基； 10 1292822 4) 生物分子，例如蛋白質，核酸碎片，聚醣、脂質、糖 蛋白、荷爾蒙、寡核酸及抗體； 5) 生物分子錯合物，例如DNA-蛋白質錯合物與蛋白質 -蛋白質錯合物； ' 6) 有機聚合物，例如聚笨乙烯及聚乙二醇； 7) 奈米級及微米級粒子，包含量子點及聚苯乙烯粒子體 積標準品； 8) 在表面的團簇或氣溶膠，特別是環境應用基材收集之 微米/奈米粒子； 9) 胞器，例如染色體或粒腺體； 10) 細胞，例如病毒、細菌和紅血球； 11) 類菌質體；及 12) 金屬與無機團藤（clusters)及粒子。 其他類型的分子也可被聲波脫附，且被提供於本發明 所述之質譜分析。 回到第一圖與第二圖中，聲波源109係設置用以造成 利用一稱做雷射誘導聲波脫附（Laser-Induced Acoustic Desorption，LIAD)技#f將樣品105之分子的聲波脫附。如 圖所示，聲波源109包含一電磁波源，例如一脈衝紫外光 雷射106,以提供基材104能量誘發基材内的聲波。一合適 的脈衝紫外光雷射106係為一頻率兩倍於Nd : YAG的雷 射。聲波源109也包含基材104，樣品105係設置於基材之 表面藉以接近進入QIT 102的開口處。雷射光束可利用聚焦 鏡107聚焦於基材上。雷射1〇6的光束係導引至基材未設 置樣品105之側。因此，樣品1〇5並無直接與雷射光束接 1292822 觸，且其可因此免於因直接暴露於雷射輻射造成之傷害。 弟二圖係為一利用雷射誘導聲波脫附技術之分子脫附 程序的示意圖。如圖所示，由雷射源1〇6來的雷射光束310 直擊設置有樣品105的基材1〇4的表面320,且可得知，樣 品105係設置於並未直接與雷射光束31〇接觸之表面322。 雷射的強度所致之光束流（即雷射能量密度）係高於 剝落門檻（即材料吸收雷射能量能成功打斷材料之分子間 鍵結的能量點）。被吸收的雷射光束因此造成物質排列於 基材上的鍵結被破壞。如第一圖之插圖所示，其係為 基材104之表面320上的半徑接近1 mm的雷射剝離點。 當物質與基材104間的鍵結被破壞後，產生的衝擊波 穿過基材直到抵達表面322。穿透波的能量傳遞到樣品1〇5 中的至少部分分子。於是能量被分子獲得後造成部分分子 由樣品105主體被脫附。由第三圖進一步來看，部分被脫 ^的分I包含中性分子（以白色圓呈現），而部分被脫附 分帶，荷分子（以黑色圓呈現）。被脫附分子中只有 部^帶電荷分子會被一離子阱質譜儀的電場捕捉 ，同時中 性分子並不會被電場影響。儘管圖未顯示，一聲波換能器 可用來監測聲波的成果。例如，可監測聲波脫附程序。 ―為了促進基材内震動波的產生，基材104係由具有一 剝落門檻低於所使用之雷射強度的材料所構成。用來做為 基材的&適材料係為;g夕，也可使用其他具有適合之剝落門 檻或其他可令其適合誘導材料内震動波特性之材料。 在部分實施態樣中（圖未呈現）’聲波源109包含-粒子束產生器（例如—電子束）。粒子束誘發基材内聲波 12 1292822 使樣品105脫附分子。以雷射光束為例，當光束流程度超 於基材剝落門檻值時，粒子束對基材上的輻射造成基材内 物質的破壞。其結果係使基材内產生震動波並穿透基材。 當震動波抵達沈積之樣品之表面時，震動波脫附至少部 的分子。 在部分實施態樣中（圖未呈現），聲波源1〇9包含一 可造成機械震動之壓電元件。在這些實施態樣中，一控制 器發出電子訊號至壓電元件，造成壓電元件根據訊號強度 啟動元件發生機械位移。壓電元件係位於接近相對於樣品 分子沈積之基材表面之相對表面附近。當壓電元件機械Z 移時，其打擊基材或基盤，因此造成聲波或震動波的產生 並穿透基材。這些穿透波抵達基材上沈積有分子樣品之表 面時造成至少部分分子由樣品中被脫附。可被啟動以產生 機械震動並傳遞至基材之其他類型的元件也可被使用。 在部分實施態樣中，聲波產生器可用以產生直接投射 至樣品上的聲波。因此，產生的聲波無需穿透其他介質， 且聲波的產生並未透過入射光束（粒子束或光束）誘發基 材内震動波的中間程序，或者產生機械震動以製造基材内 振動。在部分實施態樣中，聲波產生器係可為連續音波源、 超音波源及或脈衝聲波源。因此產生的聲波被投射至設置 於容器或基材上之分子樣品。當聲波擊中樣品，其傳遞聲 波能量至分子，其結果係至少部分分子獲得足夠動能驅使 其由樣品中脫附。至少部分被脫附的分子為帶電荷分子， 因此那些分子可進行質譜分析。在部分實施態樣中，被脫 附分子將因此具有一與分子樣品105内實質上相同之電荷 13 1292822 分佈。 儘管此處所述之聲波脫附技術的使用並不需要離子化 程序用將分子樣品105離子化。在部分情況下，樣品的離 子化仍然可以實行。舉例來說，在部分例子中需要較大量 的帶電荷分子。例如，當聲波脫附係藉由直接投射入射聲 波能量至樣品105 (例如使用連續聲波源、超音波源及/或 脈衝聲波源）來實行時，可進一步利用一般離子技術進行 離子化。一種此類離子化技術係利用一電子槍產生一電子 束並導引至分子以產生帶電荷分子，其他使樣品分子帶電 荷的方法包含使用可產生碰撞程序或光離子化程序的裝 置、可產生光子誘發電荷轉移的裝置、可產生電子貼附離 子化技術的裝置、可產生離子貼附離子化技術的裝置等。 在部分實施態樣中，聲波輻射本身造成樣品中的分子變成 離子狀態。樣品105之分子的離子化可在進行施用聲波能 量以脫附分子之前、過程中或之後。 回到第一圖中，被脫附的帶電荷粒子被導引至質譜量 測與偵測設備119中之QIT 102。QIT 102可為任何市售之 QIT質譜分析器，例如：Jordan C-1251 QIT。QIT 102商品 藉由包含環狀電極128與末端罩狀（end-cap)電極127a與 127b數個電極形成的三維四極電場捕捉帶電荷分子進入一 振盪執道（oscillatory trajectory)。導引帶電荷分子進入阱 中的額外運動係依據施加電壓、驅動頻率及被捕捉分子特 有的質荷比數值（儘管係以單一分子或粒子為參考，但可 理解的是QIT 102内部可具有多過一個的分子）。因此，當 玎明顯知悉如下··一被捕捉之帶電荷分子的質荷比可基於 14 1292822 其運動及QIT施加的電壓與驅動頻率來偵測。 被脫附的樣品分子可由環狀及末端罩狀電極間的缺口 或由環狀電極的孔中導入阱中。為確保帶電荷分子進入qIT 後保持在其中，一緩衝氣體在帶電荷分子通過QIT 1〇2時將 ▼笔荷分子之運動減緩。一種此緩衝氣體的代表係為氦 氣，其係使QIT 102内的壓力保持在大約1 mTorr。提供其 他類型的氣體及/或其他類似的減幅技術之減幅介質也可促 進QIT 102内部捕捉帶電荷分子。 參考第二圖，一旦一帶電荷分子15〇抵達qIT 1〇2，即 供應一交流電壓源120以於QIT 102内部創造出一電場來捕 捉該帶電荷分子150形成振盪運動。 由第二圖所示，交流電壓源12〇包含一驅動振盪器， 藉以產生具有一可調振幅及/或一可調頻率之電壓。舉例說 明，驅動振盪器122可為一合成功能產生器，藉以產生具 有在音頻及無線電頻路範圍内之頻率（即1〇〇 Hz-2M Hz) 之正弦曲線電壓訊號，以及可調之振幅程度。由驅動振盪 器122產生的電壓訊號可由一具處理器之元件自動控制。 由驅動振盪器122產生的訊號其頻率及振幅可額外的及/或 選擇性由使用者手動控制。 與驅動振盪器122連結的是一驅動變壓器126輸入終 細之功率放大器124 (power amplifier)，具可調振幅與頻 率的電壓訊號1/^係因此在變壓器126之輸出終端產生。這 些輸出終端係與末端罩狀電極127&與127b連結。當可被理 解的是其他類型的電氣結構也可被用來在QIT 102内部，創 造出帶電荷分子150進行質譜分析所需之電場。例如為了 工292822 4頜外的電壓被施加於末端罩狀電極127&與1271?間以抵銷 、力^可提供用來在QIT内創造出電場之各種結構的描述， ，及刼作qit的描述，舉例，在美國專利號6,777,673，標 題為^隹子阱質譜儀，，的整份文件可特此併入參考文獻中。 士當帶電荷分子150被創造於QIT 102中的電場捕捉住 可QIT 102的驅動電壓之頻率係手動地或自動地調整，直 ，與QIT 102的共振狀態被建立。當其因此提供驅動電壓訊 唬之驅動頻率Ω與徑向頻率（即帶電荷分子與阱1〇2的 振盪頻率）之比係為一整數值w，且觀測到帶電荷分子的徑 p軌道形成一穩定圖案時。此一圖案係為嵌入於第一圖與 ，、圖中之生狀圖案。星狀圖案的分支數目η等於驅動電 壓頻率與離子化分子之徑向頻率的比，因此，在共振狀態 下，Ω =nc〇r 〇 、〜 在美國專利申請號11/134,616的文件中有更多詳細的 午，可被整份併入參考文獻中。這些被觀測到的特性，例 ^ =狀圖案分支的數目„與粒子的質荷比、頻率、以及驅 電壓的振幅相關，因此當獲得帶電荷分子150與QIT 102 的共振狀態時，可因此測得其質荷比m/Ze。 然而’因為有無限的m與Z組合會形成相同的m/ze ’因此m/Ze值本身並無法提供明確關於粒子15〇的質量 ^的資訊。所以有一種決定分子15〇質量的方法，就是讓相 :的分子150製造出不同的電荷狀態，因此相同的分子15〇 曰產生不同的m/Ze值。既然產生的那些m/Ze值的分子150 $ =仍然相同，則就有可能基於多數被生成的值與多 電荷狀態的相關性來決定出質量所。 16 1292822 為產生可於後續決定質量的分子15〇的多數的帶 狀態’可利用-附加電荷模組，暴—第― : 所示之電子槍⑽。電子搶⑽能由―例如熱鶴絲產生U 束放射，電子束被導引穿過—末端罩狀電極ma，咖其 :之-上的其中之-孔洞。電子束擊中分子i5G並誘發^ 子150在電何狀悲的改變。附加電荷模组係可 類型的可雜分子丨㈣同電荷㈣的元 门 附加電荷模組⑽的㈣可包含—產 j 之後在研究時導引贿射至分子。 件 一旦分子150的電荷狀態被改變後，在⑽搬 場控制而在徑向軌道移動的分+ 15〇冑失去 案。因此當分子軌道變得不彳塞t f〜 ° 壓訊號之_頻率重新調整驅動電 達到共振狀態。 +的新屯何“對應φΤΚ)2 為了將分子150的軌道圖案視覺化 QIT 102的驅動頻率以使粒子15〇 错此7捫登 宰，需使用-用來以❹Γ 相穩定的執道® 案而使用用末產生放射光的光源。當協調且單色的光， t ?雷射產生的光投影至粒子時，其可能可利用合適的 摘測益觀測到散射強度隨時間波動。因此，可觀測到一例 如分子15〇的粒子隨時間的運動。因此，如第一圖所示， -光源11G以協調單色光照射分子請。合適的光源係為雷 導子Λ射，分子150散射的光接著被光學鏡 』罨何耦合兀件照相機（charge coupled dev1Ce ’ CCD )之光子捕捉元件丨〗4。一 未顯示）#、減子敵元件114連結，散 1292822 射的光，因此顯示分子的徑向執道運動。根據顯示的執道 圖木^可刼作由電壓源120產生之驅動電壓訊號之驅動頻 =的調整。當顯示元件顯示一穩定執道圖案，則可觀測的 斗寸〖生例如知疋星狀圖案的分支數目則被紀錄並使用來確 定分子150的m/Ze值。 測到粒子150數個電荷狀態各自的m/Ze值後，分 ^ 5量值可利用例如描述於美國專利公開號i lm4,6i6 Ϊ 2單+地說，該程序反覆地為分子150製造的 (即指定）不同的質荷比值。該程序接 偏差的-系所獨立計算出咖 平均質旦=: 值’以及獨立計算其選出系列的 从二工里 支忖領域熟知此項技藝當可理解豆他由帶電 術也可被使用。 祕來_帶電荷分子質量的技 偵’則刀子15〇質量以及因此I監定該分子的@库可由 具處理器元件（圖未呈現)處理。此一d::: 含-電腦及/或其他類型之適合多種應用二處 此類7L件可包含暫時或非暫時記憶 - 出功能的周邊元件。此補、套开心入二及啟用輸入/輸 ^ ^ 頰周邊兀件包含例如CD-ROM槽及/ ，人’、：5網路連接係為了下載包含電腦摔作i兒明 以啟動具處理器元件之一般操作的 二 150質量的軟體工具程式。此-具處理器元件; 用來偵測分子150質詈，式去ϋ效人m +卞j马專門厭格 4楚一 错1 或者/、了 I合用來實施其他功能。 σ ^s〜圖所示的四極質譜儀只是可用來偵測 由樣品105中利用聲波輻射脫附的分子150的質譜分析的 1292822 狀，巧的質譜儀。以其他方式設置的其他類型的質譜儀 衣飛列如不同偵測模組等也可被使用。 插 貝 口曰儀-of-flight mass spectrometer)係為一 域’類型’飛時質譜儀利用穿過一漂移區 場加球=過時間來分辨帶電荷分子間的不同質量。一電 域，並Φ有的離一個^冑能進入一個無電場飄移區 二二Γ係，帶電荷分子的電荷且v係為施加電壓。既 且有二古的動能係等於廣2/2，較輕的分子將比較重的分子 ;。因又：的占’且較快抵達位於飄移區域末端的偵測 ί量，並因=2:特殊粒子的飛行時間可偵測分子的 孫炎所描述的另一種可與聲波韓射程序結合的質言並儀 1一、羽型場式質％儀，在這種類型的質八 =-電場而被加速，接著使用一可調電 :ΐ;ίί管中的方向偏轉。只有具有相同離心力與向心； i力=㈡等Γ電荷分子將不會被偵測到。接ί: 份被決定。因此’帶電荷分子的質量與= 還有另一種也可被使用的質譜儀係為傅立 動移動。 19 1292822 被激發的被離子化分子的迴旋加速運動接著被接受哭平板 谓測獲得包含各種被僧測被離子化分子的所有被^迴旋 加速頻率之時，磁域訊號。接著操作對時間磁域訊號進行 傅立葉轉換以獲仔間％域sfl號的頻率場域圖像，傅立葉 轉換的結果被轉換成可鑑定各種被研究的帶電荷分子之/質 譜。 、 其他類型的質譜儀包含離子泳動質譜儀分析及其他類 型的離子阱負碏儀等，也可被用來與本發明所描述之聲波 脫附技術結合。 為決定分子質荷比值的帶電荷分子偵測可利用合適的 偵測器及/或偵測技術，這些包含具有第二電子喷射的 荷粒子元件，例如微管道平板（microchannelplate)、通道 倍增器（ch麻ltr〇n ) 5戈電子倍增元件（electr〇n m 如 device)，基於能量量測的债測器，例如電荷感應元件。 於貝！改變的_器’例如微懸臂 其他類塑的細。部分 儀’例如’基於光散射技術的偵測器適合 用於M四極離子阱質譜儀的粒子偵測。 實驗： 一圖分析中以聲波脫附的效果’使用一類似第 生物粒子了 1斤=備的質譜儀設備來分析一單一全細跑 矽晶圓）。兮择桦囷樣品被置於一半導體基材（0.5 mm厚 一摻斂* Μ樣品並未置入間質化合物（ matrix compound ) 〇 w⑽石摘石⑽YAG)雷射光束投射至基^背側)。 20 1292822 田射波長係使用532 nm，且其雷射能量大約每雷射脈衝3〇 ^+既然雷射光束被投射至具有樣品的基材背側（即非直 ΪΙΐ樣品本身）。脫附程序主要由雷射光誘發基材產生的 這成。接著，由樣品脫附的分子被一四極離子阱捕 從。 +氬離子雷射光束被用來照射被捕捉細胞以產生散射 ^射光。氬離子雷射的波長係使用488 nm，雷射能量約〜1〇〇 使用一光學鏡頭來加強散射雷射光的收集，然後利用 ;ϋ CCD元件的光子偵測器來制。藉著調整捕捉驅動 須^來埠成細胞的共振移動，可獲得被脫附細胞的質荷 比1被脫附分子的質量可利用電子搶來改變分子的帶電荷 ，態，並提供描述於美國專利申請號11/134,616專利的質 置決矣程縣計算分子的。儘子搶削來改變那 些被捕捉在QIT中的分子的帶電荷狀態，除此之外離子化 或帶電荷程序並未被施用於分子。因此，被導引至听龙1292822 IX. Description of the invention: [Technical field to which the invention pertains] More specifically for mass spectrometry, the present invention relates to mass spectrometry to produce gas phase charged molecules. [Prior Art] Mass spectrometry can be based on its mass-to-charge ratio (usually expressed in Saki or ▲), so it is necessary to investigate the charge molecular properties in an electric field when performing f-spectrum analysis. The nature of charged molecules can be judged by their mass to charge ratio. For example, in a quadrupole ion-trap mass spectrometer, charged molecules are trapped by an ion trap, and an applied electric field is applied to the trapped molecules to cause the molecules to operate in a manner, thereby indicating the mass-to-charge ratio (in w/ Name or representation). By detecting the mass-to-charge ratio of the captured molecule, the mass of the molecule can be detected, thereby identifying the molecule. In order to generate charged molecules for mass spectrometry, techniques commonly used to ionize molecules by mass spectrometry have been investigated, and charged molecules in at least a portion of the gas phase are provided. Techniques for ionization and generation of gas phase molecules have been explored, such as Matrix Assisted Laser Desorption Ionization (MALDI), which may cause matrix molecules in a sample to be introduced into a gas phase molecule. In addition, the ionization process often produces fragmentation and partial destruction of the target molecule. Performing an ionization procedure on the sample of the molecule often increases the complexity of the mass spectroscopic image. SUMMARY OF THE INVENTION In one aspect, the method of fabricating a gas phase molecule of the present invention comprises providing a sample of a molecule 5 1292822 having a unique charge distribution and directing a sound wave to a sample of the molecule for removal from the sample At least a portion of the molecule is attached such that the desorbed molecule has a substantially identical charge distribution as in the molecular sample. Embodiments may include one or more of the following: Directing acoustic radiation includes providing energy to a substrate to induce acoustic waves in the substrate. The sample is placed on a substrate. In some embodiments, the substrate comprises Shi Xi. The sample is placed on one surface of the substrate and the energy is provided from the other side of the substrate. Providing substrate energy includes generating electromagnetic radiation and directing the electromagnetic radiation to a surface of the substrate to induce acoustic waves within the substrate. Electromagnetic radiation can include laser radiation. Providing energy includes generating an electron beam and directing the electron beam to a surface of the substrate to induce acoustic waves within the substrate. Providing energy involves mechanically oscillating a surface of the substrate to induce acoustic waves within the substrate. For example, a piezoelectric element can be used to cause mechanical oscillations. Guiding acoustic radiation includes generating sound waves using, for example, a continuous sound source, an ultrasonic source, and/or a pulse source. The operation of directing the acoustic radiation is not simultaneous with directing the ionizing radiation to the sample. In some embodiments, the method further comprises performing mass spectrometry on the desorbed molecules, and the mass spectrometry can include time-of-flight mass spectrometry, quadrupole mass spectrometry, ion trap mass spectrometry, magnetic fan-shaped field mass spectrometry, and Fourier transform 1129822 Cyclotron resonance mass spectrometry and/or ion mobility mass spectrometry. In another aspect, the apparatus for producing a gas phase molecule of the present invention comprises a container containing a sample of a sample having a unique charge distribution; and an acoustic wave source configured to guide the sound wave Radiation to a sample of the molecule to cause at least some of the molecules in the sample to be desorbed. The desorbed molecule has a charge distribution substantially the same as in the sample of the aforementioned molecule. In another aspect, the method of fabricating a gas phase molecule of the present invention comprises providing a sample of one molecule, generating sound wave radiation using a continuous sound source, an ultrasonic source, and/or a pulse source; and directing the sound wave radiation to the foregoing A sample of the molecule is used to desorb at least a portion of the molecules in the sample. Acoustic radiation is injected into the sample of the molecule. The method also includes ionizing the aforementioned molecules. In still another aspect, the gas phase molecular device of the present invention comprises a container for accommodating a sample of one molecule; and a sound wave radiation generator configured to direct sound wave radiation to the molecule The sample is caused to cause at least some of the molecules to be desorbed from the sample. The acoustic radiation generator includes, for example, a continuous sound source, an ultrasonic source, and/or a pulse source. In still another aspect, the method of fabricating a gas phase molecule of the present invention comprises: providing a sample of one molecule; and directing a sample of the acoustic wave to the molecule to desorb at least a portion of the molecule from the sample without the ionization procedure of the sample. . The at least partially desorbed molecule has a charge. The desorbed molecules were subjected to mass spectrometry. In still another aspect, the gas phase molecular device of the present invention comprises a container for accommodating a sample of one molecule; and an acoustic wave source configured to direct sound waves to the sample of the molecule to cause at least Some of the molecules are desorbed from the sample. The at least partially desorbed molecule has a charge. A 7 1292822 ^ spectrometer can be set up to analyze the aforementioned desorbed molecules. The aforementioned samples were not exposed to sonic radiation under an ionization procedure. The details of one or more implementations of the invention are set forth in the drawings. Other features, objects, and advantages of the invention are apparent from the following claims, claims and claims. [Embodiment] The first figure is a front view of an exemplary apparatus 1 using a mass spectrometer to analyze molecules that are desorbed by acoustic radiation (i.e., released from a sample provided). The second figure is a block diagram of the first figure device (for the purpose of the simplified drawing, some of the parts in the first figure are not shown in the second figure). As shown in the first and second figures, the device 10A includes an acoustic wave source 109 that directs sonic energy to a sample 忉5 of one molecule to desorb at least a portion of the molecules in the sample. The desorbed molecules thus form gas phase molecules which are analyzed by the mass spectrometry and detection device 119 of the device. It will be readily understood that the at least partially desorbed molecule has a charge and, further, in some embodiments, the charge distribution of the desorbed molecule is substantially the same as the charge distribution in sample 105. To produce a gas phase molecule, a container 105 containing a sample of molecules is required. The sample 105 of the molecule is placed in proximity to the mass spectrometer device 119 so that a portion of the molecules can be introduced into the mass spectrometer measurement and detection device U9 after being released. In the first figure, the sample is provided with a quadrupole ion trap 102 (the open end of the pipe or conduit of the Quadmpole Ion Trap (QIT) (not shown), thereby introducing the inner region 103 of the qIT 1〇2, the quadrupole ion trap The inner region 103 of 1〇2 is the point of mass spectrometry operation. 8 1292822 The acoustic radiation produced by the acoustic source 1G9 is then directed onto the sample 105 of the molecule. The molecules in the partial sample 105 then gain sufficient kinetic energy to enable The bulk of the sample 105 is desorbed or released. Those desorbed molecules are directed into the inner 1〇3 region of the QIT 102. The gas phase molecules that are mass spectrometrically analyzed by sonic light are generally not ionized. Any molecule that is reduced in supply will have at least some of the molecules already ionized' a this has a positive or negative charge (such molecules that do not have to pass the ionization process, ie, have a charge, sometimes called "day ^ (b_ -charge) Molecule. In fact, there is also the nature of the sample. It is considered that there is no knife to take the mouth and even the high-eight sons in the ionized sample, which makes the concentration of the charge molecules change extremely momentarily, but in all samples. : two have The sub-molecular system carries a charge. Therefore, any fraction that is provided has a unique charge distribution. The knife is in the mouth when the sound wave is applied to a sample of one molecule, and the sound is occluded, thus causing the molecule. From the laser-induced acoustic desorption in the sample (10) released ===, the induced wave (4) ^ disk vibration frequency is approximated by the acoustic wave induced range, therefore, the Ray (four) wave can effectively destroy the molecular table (10) junction. , ^ induces the consistency between the frequency of the acoustic wave and the vibration frequency of the surface bond, and the transmission of the energy can be avoided, thus avoiding the loss or failure of the energy conversion, which is born in the common technology of manufacturing gas phase molecules. Further, the sound wave ^ Hanging, the molecules are detached smoothly without splitting. In some implementations, the desorbed molecules of the sample I = have the same identity as the molecules in the sample. "The charge is divided into 9 1292822, although acoustic wave desorption is used without accompanying Any other ionization may only produce a small amount of gas phase molecules. However, for mass spectrometry purposes, even a small amount of charged gas is sufficient to form a gas phase because the mass spectrometer device requires only a small amount. Charged molecules are sufficient to identify the relative w々 values of these molecules. Although the ratio of charged ions to neutral molecules/particles is low, in some cases, the overall detection efficiency is comparable to that of mass spectrometers using ionization devices. Approximate, even better. Mass spectrometry of biomolecules or biological particles in mass spectrometers using ionization devices, the ionization rate is generally very low, for example when using MALDI or electrospray ionization (ESI) The ionization efficiency of biomolecules is generally lower than 0.0001. For some large biomolecules, the efficiency may be close to 0. For example, macroglycan molecules (M> 100,000 Daltons) cannot be ionized properly using a general ionization device. Furthermore, although the unbonded molecules are released into the gas phase smoke due to sonic desorption, the presence of uncharged molecules in the gas phase does not affect the mass spectrometry results because the uncharged molecules are not charged. The particle detector detects that, in addition, the uncharged molecules are not captured by the ion trap device. The sonic desorption described herein can be applied to any type of molecule, including: 1) liquid samples and amorphous materials that do not have sufficient gas pressure at the operating temperature of the mass spectrometer, these liquid and amorphous materials contain liquid crystals and ionic liquids. (ionic liquids); 2) an analyte captured by a solid state sample, such as nitrogen captured in a metal foil, 3) an adsorbate on the surface of the substrate, such as a free radical on a catalytic metal surface; 10 1292822 4) Molecules such as proteins, nucleic acid fragments, glycans, lipids, glycoproteins, hormones, oligonucleic acids and antibodies; 5) biomolecular complexes, such as DNA-protein complexes and protein-protein complexes; '6) organic Polymers such as polystyrene and polyethylene glycol; 7) nanoscale and micron-sized particles containing quantum dots and polystyrene particle volume standards; 8) clusters or aerosols on the surface, especially for environmental applications Micron/nanoparticles collected by the substrate; 9) organelles, such as chromosomes or granulocytes; 10) cells, such as viruses, bacteria and red blood cells; 11) mycoplasma; and 12) gold With inorganic Dando (Clusters), and particles. Other types of molecules can also be desorbed by sonication and provided in the mass spectrometric analysis of the present invention. Returning to the first and second figures, the acoustic wave source 109 is arranged to cause the sound waves of the molecules of the sample 105 to be desorbed by a laser-induced Acoustic Desorption (LIAD) technique #f. . As shown, acoustic source 109 includes an electromagnetic wave source, such as a pulsed ultraviolet laser 106, to provide energy for substrate 104 to induce acoustic waves within the substrate. A suitable pulsed ultraviolet laser 106 is a laser having a frequency twice that of Nd:YAG. The acoustic wave source 109 also includes a substrate 104 that is disposed on the surface of the substrate to approximate access to the opening of the QIT 102. The laser beam can be focused onto the substrate using a focusing mirror 107. The laser beam of 1 〇6 is directed to the side of the substrate where sample 105 is not disposed. Therefore, sample 1〇5 is not directly in contact with the laser beam 1292822, and it can thus be protected from damage caused by direct exposure to laser radiation. The second diagram is a schematic diagram of a molecular desorption procedure using laser-induced sonic desorption techniques. As shown, the laser beam 310 from the laser source 1〇6 strikes the surface 320 of the substrate 1〇4 provided with the sample 105, and it can be seen that the sample 105 is disposed not directly with the laser beam. 31〇 contact surface 322. The beam current (i.e., the laser energy density) due to the intensity of the laser is higher than the spalling threshold (i.e., the energy point at which the material absorbs the laser energy to successfully break the intermolecular bond of the material). The absorbed laser beam thus causes the bond of the substance to be arranged on the substrate to be destroyed. As shown in the inset of the first figure, it is a laser peeling point having a radius of approximately 1 mm on the surface 320 of the substrate 104. When the bond between the substance and the substrate 104 is broken, the resulting shock wave passes through the substrate until it reaches the surface 322. The energy of the penetrating wave is transmitted to at least a portion of the molecules in sample 1〇5. Thus, when the energy is obtained by the molecule, part of the molecules are desorbed from the main body of the sample 105. As further seen from the third figure, the fraction I that is partially removed contains a neutral molecule (presented by a white circle), and a part is desorbed, and a charged molecule (presented by a black circle). Only the charged molecules in the desorbed molecules are captured by the electric field of an ion trap mass spectrometer, and the neutral molecules are not affected by the electric field. Although not shown, a sonic transducer can be used to monitor the results of sound waves. For example, the sonic desorption procedure can be monitored. - In order to promote the generation of shock waves in the substrate, the substrate 104 is composed of a material having a peeling threshold lower than the laser intensity used. The & materials used as the substrate are used; other materials having suitable peeling thresholds or other properties that make them suitable for inducing shock wave properties in the material may be used. In some implementations (not shown) the acoustic source 109 comprises a particle beam generator (e.g., an electron beam). Particle beam induces acoustic waves in the substrate 12 1292822 Desorbs the sample 105. Taking a laser beam as an example, when the beam current exceeds the substrate peeling threshold, the particle beam causes damage to the material in the substrate by the radiation on the substrate. As a result, shock waves are generated in the substrate and penetrate the substrate. When the shock wave reaches the surface of the deposited sample, the shock wave desorbs at least some of the molecules. In some implementations (not shown), the acoustic source 1〇9 contains a piezoelectric element that causes mechanical shock. In these embodiments, a controller emits an electrical signal to the piezoelectric element, causing the piezoelectric element to mechanically displace the element based on the signal strength. The piezoelectric element is located near the opposite surface of the surface of the substrate deposited relative to the sample molecules. When the piezoelectric element is mechanically moved Z, it strikes the substrate or the substrate, thereby causing generation of sound waves or shock waves and penetrating the substrate. These penetrating waves reach the surface of the substrate where the molecular sample is deposited causing at least some of the molecules to be desorbed from the sample. Other types of components that can be activated to generate mechanical shock and transfer to the substrate can also be used. In some implementations, the acoustic wave generator can be used to generate acoustic waves that are projected directly onto the sample. Therefore, the generated sound waves do not need to penetrate other media, and the generation of sound waves does not pass through the incident beam (particle beam or beam) to induce an intermediate process of vibration waves in the substrate, or mechanical vibration is generated to create vibration in the substrate. In some implementations, the acoustic generator can be a continuous sonic source, an ultrasonic source, or a pulsed acoustic source. The resulting sound waves are then projected onto a molecular sample placed on a container or substrate. When a sound wave hits a sample, it transmits sonic energy to the molecule, with the result that at least some of the molecules gain sufficient kinetic energy to drive them out of the sample. At least partially desorbed molecules are charged molecules, so those molecules can be analyzed by mass spectrometry. In some embodiments, the desorbed molecule will thus have a substantially identical charge 13 1292822 distribution within the molecular sample 105. Although the use of sonication techniques described herein does not require ionization procedures to ionize molecular sample 105. In some cases, the ionization of the sample can still be carried out. For example, a larger amount of charged molecules are required in some examples. For example, when sonic desorption is performed by directly projecting incident acoustic energy into the sample 105 (e.g., using a continuous acoustic source, an ultrasonic source, and/or a pulsed acoustic source), ionization can be further performed using conventional ion techniques. One such ionization technique utilizes an electron gun to generate an electron beam and direct it to a molecule to produce charged molecules. Other methods of charging a sample molecule include the use of a device that produces a collision procedure or photoionization procedure that produces photons. A device for inducing charge transfer, a device capable of generating an electron attaching ionization technique, a device capable of generating an ion attached ionization technique, and the like. In some implementations, sonic radiation itself causes the molecules in the sample to become ionic. Ionization of the molecules of sample 105 can be performed before, during, or after application of sonic energy to desorb molecules. Returning to the first figure, the desorbed charged particles are directed to the QIT 102 in the mass spectrometry and detection device 119. The QIT 102 can be any commercially available QIT mass spectrometer, such as the Jordan C-1251 QIT. The QIT 102 commercial captures charged molecules into an oscillatory trajectory by a three-dimensional quadrupole electric field comprising a ring electrode 128 and a plurality of electrodes of end-cap electrodes 127a and 127b. The additional motion that directs the charged molecules into the trap depends on the applied voltage, the drive frequency, and the mass-to-charge ratio value specific to the captured molecule (although reference is made to a single molecule or particle, it is understood that there may be more inside the QIT 102) Pass one molecule). Therefore, it is apparent that the mass-to-charge ratio of a captured charged molecule can be detected based on the motion and the voltage applied by the QIT and the driving frequency of 14 1292822. The desorbed sample molecules can be introduced into the well from the gap between the annular and end cap electrodes or from the holes of the ring electrode. To ensure that the charged molecules remain in the qIT and remain there, a buffer gas slows the movement of the gun molecules as the charged molecules pass QIT 1〇2. One such buffer gas is represented by helium which maintains the pressure within the QIT 102 at about 1 mTorr. A moderating medium that provides other types of gas and/or other similar reduction techniques can also facilitate the capture of charged molecules within the QIT 102. Referring to the second figure, once a charged molecule 15 〇 reaches qIT 1〇2, an AC voltage source 120 is supplied to create an electric field inside the QIT 102 to capture the charged molecules 150 to form an oscillating motion. As shown in the second figure, the AC voltage source 12A includes a drive oscillator to generate a voltage having an adjustable amplitude and/or an adjustable frequency. For example, the drive oscillator 122 can be a composite function generator to generate a sinusoidal voltage signal having a frequency within the range of audio and radio frequencies (ie, 1 〇〇 Hz - 2 Hz), and an adjustable amplitude level. . The voltage signal generated by drive oscillator 122 can be automatically controlled by components of a processor. The frequency and amplitude of the signal generated by drive oscillator 122 can be additionally and/or selectively controlled manually by the user. Connected to the drive oscillator 122 is a drive transformer 126 that inputs a final power amplifier 124. The voltage signal 1/^ with adjustable amplitude and frequency is thus generated at the output terminal of the transformer 126. These output terminals are connected to the end cap electrodes 127 & 127b. It will be appreciated that other types of electrical structures can also be used within QIT 102 to create the electric field required for mass spectrometry of charged molecules 150. For example, in order to work 292822, the voltage outside the jaw is applied to the end cap electrode 127 & and 1271 to offset, the force can provide a description of the various structures used to create an electric field in the QIT, and the qit The entire document, which is described in U.S. Patent No. 6,777,673, the disclosure of which is incorporated herein by reference. The charge-carrying molecule 150 is created by the electric field created in the QIT 102. The frequency of the drive voltage of the QIT 102 is manually or automatically adjusted, and the resonance state with the QIT 102 is established. When it thus provides a driving voltage signal, the ratio of the driving frequency Ω to the radial frequency (ie, the charged molecule and the oscillation frequency of the well 1〇2) is an integer value w, and the formation of the path p of the charged molecule is observed. When a pattern is stabilized. This pattern is a green pattern embedded in the first figure and the figure. The number of branches η of the star-shaped pattern is equal to the ratio of the frequency of the driving voltage to the radial frequency of the ionized molecules, and therefore, in the resonance state, Ω = nc 〇 〇 , 〜 in the document of US Patent Application No. 11/134,616 More detailed noon can be incorporated into the reference in its entirety. These observed characteristics, the number of branching patterns of the pattern = are related to the mass-to-charge ratio of the particles, the frequency, and the amplitude of the driving voltage, so when the resonance state of the charged molecule 150 and the QIT 102 is obtained, it can be measured The mass-to-charge ratio m/Ze is obtained. However, 'because there are infinite m and Z combinations, the same m/ze will be formed. Therefore, the m/Ze value itself does not provide information on the quality of the particle 15〇. Therefore, there is a kind of information. The method of determining the mass of a molecule is to let the phase 150 of the phase produce different states of charge, so that the same molecule 15〇曰 produces different m/Ze values. Since the molecules of those m/Ze values are generated 150 $ = Still the same, it is possible to determine the mass based on the correlation between the majority of the generated values and the state of the multi-charge. 16 1292822 The state of the band that produces a majority of the molecules that can be subsequently determined by the mass 15 ' Module, Storm - No.: The electron gun (10) shown. The electronic grab (10) can generate U beam radiation from, for example, hot crane wire, and the electron beam is guided through the end cap electrode ma, on the - One of them - the hole. Hits the molecular i5G and induces a change in the electrical conductivity of the sub-150. The additional charge module is of the type of heterogeneous molecule (4) and the charge (4) of the elementary gate additional charge module (10) can be included - after the production During the study, the bribe was directed to the numerator. Once the charge state of the numerator 150 was changed, it was lost in the (10) shift control and moved in the radial orbital + 15 。. Therefore, when the molecular orbit became uncongested tf~ ° The frequency of the pressure signal re-adjusts the drive power to reach the resonance state. + The new geometry "corresponds to φ ΤΚ" 2 In order to visualize the orbital pattern of the molecule 150 to drive the frequency of the QIT 102, the particle 15 is wrong. Need to use - to use a source that produces a radiant light with a stable phase. When the coordinated and monochromatic light, the light produced by the t-ray is projected onto the particle, it may be possible to observe the scattering intensity fluctuating with time using a suitable sweeping benefit. Therefore, an example of the movement of particles such as molecules 15 随 over time can be observed. Therefore, as shown in the first figure, the light source 11G illuminates the molecules with coordinated monochromatic light. A suitable source of light is a ray-directed ray, and the light scattered by the numerator 150 is then optically mirrored by a photon capture element charge4 of a charge coupled dev1Ce ’ CCD. One does not show) #, minus the enemy element 114 is connected, scattered 1292822 light, thus showing the radial orbital motion of the molecule. According to the displayed trajectory, the driving frequency of the driving voltage signal generated by the voltage source 120 can be adjusted. When the display element displays a stable cue pattern, the number of branches of the observable cell, such as the known star-like pattern, is recorded and used to determine the m/Ze value of the molecule 150. After measuring the respective m/Ze values of the plurality of charge states of the particle 150, the value of the fraction can be manufactured for the molecule 150, for example, as described in U.S. Patent Publication No. lm4,6i6 Ϊ 2 (ie specify) different mass-to-charge ratio values. The program is independent of the calculation of the average quality of the denier =: value 'and the independent calculation of its selected series from the second division of the field of support is well known in the field. It can be understood that the bean can also be used by electrification. The secret _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ This d::: contains - computer and / or other types suitable for a variety of applications. Such 7L pieces can contain peripheral components that are temporary or non-temporary memory - out of function. This supplement, set up happy into the second and enable input / input ^ ^ cheek peripherals including, for example, CD-ROM slot and /, people', : 5 network connection in order to download the computer containing the computer to start the processor A two-150-quality software utility program for the general operation of components. This has a processor component; it is used to detect the quality of the molecule 150, and it is used to implement other functions. The quadrupole mass spectrometer shown in σ ^s ~ is only a 1292822-like mass spectrometer that can be used to detect mass spectrometry analysis of molecules 150 desorbed by sonic radiation in sample 105. Other types of mass spectrometers that are otherwise set up, such as different detection modules, can also be used. The Bayer-of-flight mass spectrometer is a domain type-type time-of-flight mass spectrometer that utilizes a drift zone field plus ball = time to resolve different masses between charged molecules. An electrical field, and Φ some from a ^ can enter an electric field-free drift zone, the second dioxin, the charge of the charged molecule and the v system is the applied voltage. Both the second kinetic energy system is equal to 2/2, and the lighter molecules will be heavier. Because of the :: and the faster detection of the amount of detection at the end of the drift zone, and because of =2: the special particle's time-of-flight detectable molecule Sun Yan described another combination with the sonic Korean program The qualitative statement is that the feather field type mass spectrometer is accelerated in this type of mass== electric field, and then an adjustable electric power is used: ΐ; ίί directional deflection in the tube. Only those with the same centrifugal force and centripetal; i force = (b) will not be detected. ί: The share was decided. Therefore, the mass of the charged molecule and = another mass spectrometer that can also be used is the Fourier movement. 19 1292822 The vortex-accelerated motion of the excited ionized molecule is then accepted by the cryo-plate. The magnetic domain signal is obtained when all the gyro-acceleration frequencies of the ionized molecules are detected. Next, the Fourier transform is performed on the time domain signal to obtain the frequency field image of the % field sfl, and the result of the Fourier transform is converted into a mass spectrum which can identify various charged molecules to be studied. Other types of mass spectrometers, including ion mobility mass spectrometer analysis and other types of ion trap negative oximeters, can also be used in conjunction with the sonic desorption techniques described herein. Detecting charged molecules for determining molecular mass-to-charge ratios may utilize suitable detectors and/or detection techniques, including charged particle elements having a second electron injection, such as microchannel plates, channel multipliers ( Ch heltr〇n) 5 Ge electron multiplying element (electr〇nm such as device), based on energy measurement of the debt detector, such as charge sensing components. Yu Bei! The changed _ device 'for example, the micro cantilever is similar to other plastic moldings. Some instruments, such as 'light scattering based detectors, are suitable for particle detection of M quadrupole ion trap mass spectrometers. Experiment: The effect of sonic desorption in a graph analysis uses a mass spectrometer device similar to the first bioparticle to analyze a single full-scale run wafer. The sample of the selected birch was placed on a semiconductor substrate (0.5 mm thick and one mixed * Μ sample was not placed in the matrix compound 〇 w (10) stone picking stone (10) YAG) laser beam projected onto the base back side) . 20 1292822 The field emission wavelength is 532 nm and its laser energy is approximately 3 每 per laser pulse + ^ since the laser beam is projected onto the back side of the substrate with the sample (ie the non-straight sample itself). The desorption procedure is primarily caused by laser light induced substrate formation. Next, the molecules desorbed from the sample are captured by a quadrupole ion trap. + An argon ion laser beam is used to illuminate the captured cells to produce scatter. The wavelength of the argon ion laser is 488 nm, and the laser energy is about ~1 〇〇. An optical lens is used to enhance the collection of scattered laser light, and then the photon detector of the CCD element is used. By adjusting the capture drive to obtain the resonance movement of the cells, the mass-to-charge ratio of the desorbed cells can be obtained. The mass of the desorbed molecules can be utilized to change the charge and state of the molecule, and is described in the United States. The patent application No. 11/134,616 patents the quality of the calculation of the numerator. The robbing is used to change the charged state of the molecules trapped in the QIT, except that the ionized or charged program is not applied to the molecule. Therefore, being guided to the dragon
捕捉於QIT中的分子係為經由聲波脫附程序之天生帶 分子。 J ★第四圖，為被债測之利用雷射誘發聲波脫附的大腸桿 菌全細胞群落之細胞數目（7Vc)與質量的關係圖。利用電子 微鏡製作的大腸桿菌分子的影像係如嵌入於第四圖中之的 影像，欲人影像的比例尺絲！微米，生物性全細胞分子 經常被以細胞聚落方式導引進人f譜儀，因此㈣測 質荷比對應於分子群落優於獨立分子。如第四圖所述 來偵測的脫水大腸桿菌細胞的平均質量m/Nc係為5·35 + 0.24x10運耳吞（Da)。由此可知細菌脫水前約含有7〇〜8〇% 1292822 的水。因此，此量測數據推知細胞的濕重〜0.35Pg。其係吻 合=由體〜1微米長x〜0 6微米直徑）與密度（〜H g/cm3) 估异的細菌粒子。因此，雷射誘發聲波脫附可準確地測定 質量與鑑定分子（即大腸桿菌細胞）。 一類似的質譜分析係利用聲波脫附操作於具有直徑 0·269±0·007 nm的聚笨乙烯奈米粒子。第五圖係為利用雷射 誘發聲波脫附之粒子數目對偵測到的聚苯乙烯奈米球群落 的質篁的關係圖。如第五圖所示，偵測到的平均質量為617 ±0.18xl09Da，此平均值與利用單一聚苯乙烯奈米粒子的直 ，0.369微米及後、度值1.055 g/cm3計算的6.5±0.4xl〇9Da相 當。利用電子顯微鏡製造的聚苯乙烯粒子影像圖係嵌入於 第五圖中，嵌入影像的比例尺係為300 nm 〇 苐六圖係為利用雷射誘發聲波脫附由〇·5 mm厚石夕晶圓 上釋放的粒子的電荷狀態分佈圖。被量測的兩個樣品係為 0·269 nm的聚苯乙埽奈米粒子（第六圖區域61〇所示）及 大腸桿細分子（區域620所示）。如圖所示，對於每一個聚 本乙稀奈米粒子及大腸桿菌分子而言，導入質譜儀之被脫 附的帶電荷分子具有一個對稱的分佈。這些帶電荷分子的 電荷分佈並未利用一般市售質譜儀分子離子化的程序來建 立。儘管如此’如同已提醒過的’離子化模組也可被使用 來使本發明所述之聲波脫附程序所脫附的分子帶有電荷。 第六圖係呈現聲波脫附程序可由樣品中提供天生帶電荷分 子而無需操作額外的離子化程序。 其他實施熊樣 22 本發明已描述一系列的實施態樣，在本說明書中所揭 露的所有特徵都可能與其他方法結合，本說明書中所揭露 的每一個特徵都可能選擇性的以相同、相等或相似目的特 徵所取代，因此，除了特別顯著的特徵之外，所有的本說 明書所揭露的特徵僅是相等或相似特徵中的一個例子。 雖然本發明已以較佳實施例揭露如上，然其並非用以 限定本發明，任何熟悉此技藝者，在不脫離本發明之精神 和範圍内，當可作各種之更動與潤飾。因此，其他的實施 態樣可參照後續申請專利範圍所載。The molecular system captured in the QIT is a natural band molecule via a sonic desorption procedure. J ★ Figure 4 is a graph showing the relationship between cell number (7Vc) and mass of the whole cell population of Escherichia coli that was desorbed by laser-induced sonication. The image of the E. coli molecule produced by the electron micromirror is the image embedded in the fourth image, and the scale of the image is desired! Micron, biological whole-cell molecules are often introduced into the human f spectrometer in the form of cell colonies, so (4) the mass-to-charge ratio corresponds to the molecular population superior to the independent molecule. The average mass m/Nc of the dehydrated E. coli cells detected as described in the fourth panel was 5.35 + 0.24 x 10 otox (Da). It can be seen that the water contains about 7〇~8〇% 1292822 before the bacteria are dehydrated. Therefore, this measurement data infers that the wet weight of the cells is ~0.35 Pg. Its lineage = = ~1 micron long x ~ 0 6 micron diameter) with density (~H g / cm3) to estimate the bacterial particles. Therefore, laser-induced sonic desorption can accurately determine the quality and identity of molecules (ie, E. coli cells). A similar mass spectrometry was performed using sonic desorption on polystyrene nanoparticles having a diameter of 0·269 ± 0·007 nm. The fifth graph is a graph showing the relationship between the number of particles induced by laser-induced sonic desorption and the detected quality of the polystyrene nanosphere community. As shown in the fifth graph, the average mass detected is 617 ± 0.18 x l09 Da, which is 6.5 ± 0.4 calculated from the straight, 0.369 μm and the subsequent value of 1.055 g/cm 3 using a single polystyrene nanoparticle. xl〇9Da is equivalent. The image of polystyrene particles produced by electron microscopy is embedded in the fifth image, and the scale of the embedded image is 300 nm. The six-figure system is used for laser-induced acoustic desorption by 〇·5 mm thick lithography wafer. A map of the state of charge of the particles released. The two samples that were measured were 0. 269 nm polystyrene nanoparticles (shown in the sixth panel area 61 )) and large intestine rod fine molecules (shown in region 620). As shown, for each of the polyethylene nanoparticles and E. coli molecules, the desorbed charged molecules introduced into the mass spectrometer have a symmetric distribution. The charge distribution of these charged molecules is not established using the procedures of molecular ionization of commercially available mass spectrometers. Nonetheless, as has been suggested, an ionization module can be used to charge a molecule desorbed by the sonication procedure of the present invention. The sixth figure shows that the sonic desorption procedure can provide natural charged molecules in the sample without the need to operate an additional ionization procedure. Other Implementations of Bear Samples 22 The present invention has been described in terms of a series of embodiments, all of which may be combined with other methods, and each of the features disclosed in this specification may be selectively identical and equal. The features of the present invention are replaced by features of similar nature and, therefore, all of the features disclosed in this specification are merely one of the equivalent or similar features. While the invention has been described above in terms of the preferred embodiments thereof, it is not intended to limit the invention, and various modifications and changes can be made without departing from the spirit and scope of the invention. Therefore, other implementation aspects can be found in the scope of the subsequent patent application.
1292822 23 1292822 【圖式簡單說明】 第一圖係為使用聲波脫附之質譜儀裝置範例態樣之前 視概圖。 第二圖係為第一圖裝置之區塊概要圖。 第三圖係為描述利用雷射聲波誘發脫附技術之分子脫 附程序。 第四圖係為利用雷射誘發聲波脫附之質譜儀偵測大腸 桿菌全細胞群之細胞數〇/Vc)與質量m相關性趨勢圖。 • 第五圖係為利用雷射誘發聲波脫附之質譜儀偵測聚苯 乙烯奈米珠群之粒子數與質量m相關性趨勢圖。 第六圖係為由0.5mm厚矽晶圓表面利用雷射誘發聲波 脫附分子之電荷狀態分佈圖。 圖中相同符號代表相同元件。 【主要元件符號對照說明】 100 範例裝置 102 四極離子拼 103 四極離子阱之内部區域 104 基材 105 分子之樣品 106 脈衝紫外光雷射 107 聚焦鏡 108 電子槍 109 聲波源 110 光源 24 1292822 114 光子捕捉元件 119 質譜儀設備 120 交流電壓源 122 驅動振盪器 124 功率放大器 126 變壓器 127a、127b 末端罩狀電極 150 帶電荷分子1292822 23 1292822 [Simple description of the diagram] The first diagram is a pre-view diagram of an example of a mass spectrometer device using sonic desorption. The second figure is a block diagram of the device of the first figure. The third figure is a molecular desorption procedure describing the use of laser-induced detachment techniques. The fourth graph is a graph showing the correlation between the number of cells of the whole cell population of Escherichia coli 〇/Vc and the mass m using a mass spectrometer using a laser-induced sonic desorption. • Figure 5 is a plot of the correlation between the number of particles and the mass m of the polystyrene nanobeads using a mass spectrometer that uses laser-induced sonic desorption. The sixth figure is a charge state distribution diagram of a laser-induced sonic desorption molecule from a 0.5 mm thick germanium wafer surface. The same symbols in the figures represent the same elements. [Main component symbol comparison description] 100 Example device 102 Quadrupole ion 103 Internal region of quadrupole ion trap 104 Substrate 105 Molecular sample 106 Pulsed ultraviolet laser 107 Focusing mirror 108 Electron gun 109 Acoustic source 110 Light source 24 1292822 114 Photon capture element 119 mass spectrometer device 120 AC voltage source 122 drive oscillator 124 power amplifier 126 transformer 127a, 127b end cap electrode 150 charged molecule
310 雷射光束 610 聚苯乙烯奈米粒子 620 大腸桿菌分子310 laser beam 610 polystyrene nanoparticle 620 Escherichia coli molecule
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