JP2011530082A - 細胞ミクロ構造の共焦点画像化のための染料適用 - Google Patents
細胞ミクロ構造の共焦点画像化のための染料適用 Download PDFInfo
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- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
- A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
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- G02B21/002—Scanning microscopes
- G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
- G02B21/0028—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders specially adapted for specific applications, e.g. for endoscopes, ophthalmoscopes, attachments to conventional microscopes
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- G—PHYSICS
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- G02B21/00—Microscopes
- G02B21/0004—Microscopes specially adapted for specific applications
- G02B21/002—Scanning microscopes
- G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
- G02B21/0032—Optical details of illumination, e.g. light-sources, pinholes, beam splitters, slits, fibers
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- G—PHYSICS
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- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/0004—Microscopes specially adapted for specific applications
- G02B21/002—Scanning microscopes
- G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
- G02B21/0052—Optical details of the image generation
- G02B21/0076—Optical details of the image generation arrangements using fluorescence or luminescence
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- A61B1/00—Instruments for performing medical examinations of the interior of cavities or tubes of the body by visual or photographical inspection, e.g. endoscopes; Illuminating arrangements therefor
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- A—HUMAN NECESSITIES
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- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B1/00—Instruments for performing medical examinations of the interior of cavities or tubes of the body by visual or photographical inspection, e.g. endoscopes; Illuminating arrangements therefor
- A61B1/04—Instruments for performing medical examinations of the interior of cavities or tubes of the body by visual or photographical inspection, e.g. endoscopes; Illuminating arrangements therefor combined with photographic or television appliances
- A61B1/043—Instruments for performing medical examinations of the interior of cavities or tubes of the body by visual or photographical inspection, e.g. endoscopes; Illuminating arrangements therefor combined with photographic or television appliances for fluorescence imaging
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- A—HUMAN NECESSITIES
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- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
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- A61B5/7285—Specific aspects of physiological measurement analysis for synchronising or triggering a physiological measurement or image acquisition with a physiological event or waveform, e.g. an ECG signal
Abstract
Description
本願は、2008年8月4日に出願された、米国仮特許出願第61/137876号の優先権および利益を主張し、その全体は、参照することによって本明細書に組み込まれる。
Alexa Fluor染料の光退色に対する耐性は、十分に高く、高強度レーザー源によって照射される際にさえ、蛍光強度は、色褪せ防止剤が不在の場合にさえ、ある期間、概して安定なままである。この特徴は、水溶性Alexa Fluorプローブを、生細胞および組織切片両方の調査、ならびに従来の固定試料のために容易に利用できるようにする。
一実験手順では、成体ウサギは、ペントバルビタール(30mg/kg)で麻酔され、ヘパリン(2500USP単位/kg)で抗凝血された。開胸に続き、ウサギの心臓は、素早く切除され、室温で、修飾酸素処理Tyrodeの溶液(mM単位で、126NaCl、11D形グルコース、0.1CaCl2、13.2KCl、1MgCl2、12.9NaOH、24HEPES)中に定置された。心臓は、3種類の組織切片、右心室乳頭筋(約1mm×1mm×5mm)、心外膜下心室組織(約6mm×2mm)、および心房組織(約6mm×2mm)に切離された。これらの切片は、図8に示されるように、縫合糸によってポリカーボネートホルダに固定され、画像化まで溶液中で保管された。
Claims (34)
- 組織の画像化のための方法であって、
遠位端部を有する光ファイバ束を提供することと、
染料キャリアを前記光ファイバ束の前記遠位端部に動作可能に連結することであって、前記染料キャリアは、少なくとも1つの蛍光染料を備える、ことと、
前記少なくとも1つの蛍光染料の少なくとも一部分を着目組織領域に選択的に導入するために、前記染料キャリアの一部分を前記着目組織領域に接触させて位置付けることと、
所望の波長の光を生成することと、
前記組織内で前記蛍光染料を励起するために、前記光ファイバ束の中および前記染料キャリアを通して、前記着目組織領域に前記光を伝送することと、
を含む、方法。 - 前記励起された蛍光染料から前記光ファイバ束に放射される光を受信することと、前記放射光を共焦点顕微鏡画像化システムに通信することとをさらに含む、請求項1に記載の方法。
- 前記共焦点画像を形成するように、前記受信した放射光を処理することをさらに含む、請求項2に記載の方法。
- 前記被検体から得られるECG信号を監視することであって、前記被検体から得られる前記ECG信号の基準点は、前記画像獲得をトリガする、ことをさらに含む、請求項1に記載の方法。
- 光伝送のステップは、前記ECG信号上の各連続基準点で繰り返される、請求項4に記載の方法。
- 前記光ファイバ束への各光伝送の結果として、前記励起された蛍光染料から放射される光を受信することと、前記放射光を共焦点顕微鏡画像化システムに通信することとをさらに含む、請求項5に記載の方法。
- 前記共焦点画像を形成するように、前記受信した放射光を処理することをさらに含む、請求項6に記載の方法。
- 前記染料キャリアは、光透明マトリックスと、前記少なくとも1つの蛍光染料とを備える、請求項1に記載の方法。
- 前記少なくとも1つの蛍光染料は、緩衝溶液中に懸濁される、請求項2に記載の方法。
- 前記少なくとも1つの蛍光染料および前記緩衝溶液は、前記染料キャリアの少なくとも95%を構成する、請求項9に記載の方法。
- 前記少なくとも1つの蛍光染料は、所定の濃度である、請求項8に記載の方法。
- 前記染料キャリアは、少なくとも1つの抗体剤を含む、請求項8に記載の方法。
- 前記少なくとも1つの蛍光染料は、Alexa、Texas Red、FITC、Oregon Green、Rhodamine Green、Lucifer yellow、Fluo3、Fluo4、およびジ−8−Aneppsからなる群から選択される、請求項1に記載の方法。
- 前記少なくとも1つの蛍光染料は、40kDa以下の分子量を有する、請求項1に記載の方法。
- 前記少なくとも1つの蛍光染料は、20kDa以下の分子量を有する、請求項14に記載の方法。
- 前記少なくとも1つの蛍光染料は、約3kDa〜約10kDaの分子量を有する、請求項1に記載の方法。
- 前記光透明マトリックスは、ヒドロゲルを含む、請求項8に記載の方法。
- 前記光透明マトリックスは、前記光ファイバ束の前記遠位端部から100μm未満延在する、請求項17に記載の方法。
- 前記光透明マトリックスは、前記光ファイバ束の前記遠位端部から200μm未満延在する、請求項17に記載の方法。
- 前記着目組織は、心臓組織である、請求項1に記載の方法。
- プロセッサと、所望の波長の光エネルギーを選択的に生成するように構成される、光エネルギー源とを含む、共焦点顕微鏡画像化システムにおいて、被検体の組織の共焦点画像を現像するためのカテーテルであって、
遠位端部と、反対の近位端部とを有する、光ファイバ束であって、前記近位端部は、前記共焦点顕微鏡画像化システムと動作可能な通信状態にあり、前記光ファイバ束は、前記光エネルギー源と通信状態にある、光ファイバ束と、
少なくとも1つの蛍光染料を前記光ファイバ束の前記遠位端部に位置付けるための手段と、
を含む、カテーテル。 - 前記少なくとも1つの蛍光染料を着目組織領域に選択的に導入するために、前記染料キャリアの一部分を前記着目組織領域に接触させて位置付けるための手段をさらに含む、請求項21に記載のカテーテル。
- 前記プロセッサは、前記共焦点画像を形成するように、前記少なくとも1つの蛍光染料から受信された放射光を処理するように構成される、請求項22に記載のカテーテル。
- 前記プロセッサは、前記被検体のECG信号を使用して、共焦点画像化をトリガするように構成される、請求項23に記載のカテーテル。
- 少なくとも1つの蛍光染料を前記光ファイバ束の前記遠位端部に位置付けるための前記手段は、前記光ファイバ束の前記遠位端部に連結した染料キャリアを備え、前記少なくとも1つの蛍光染料は、前記染料キャリアに装填され、前記染料キャリアは、光透過性マトリックスを含む、請求項22に記載のカテーテル。
- 前記少なくとも1つの蛍光染料は、緩衝溶液中に懸濁される、請求項25に記載のカテーテル。
- 前記少なくとも1つの蛍光染料および前記緩衝溶液は、前記染料キャリアの少なくとも95%を構成する、請求項26に記載のカテーテル。
- 前記少なくとも1つの蛍光染料は、所定の濃度である、請求項25に記載のカテーテル。
- 前記染料キャリアは、少なくとも1つの抗体剤を含む、請求項25に記載のカテーテル。
- 前記少なくとも1つの蛍光染料は、Alexa、Texas Red、FITC、Oregon Green、Rhodamine Green、Lucifer yellow、Fluo3、Fluo4、およびジ−8−Aneppsからなる群から選択される、請求項21に記載のカテーテル。
- 前記光ファイバ束の遠位端部分を選択的に、かつ少なくとも部分的に囲むように構成される、カテーテルシースをさらに備える、請求項25に記載のカテーテル。
- 前記カテーテルシースは、前記染料キャリアの少なくとも一部分を選択的に、かつ少なくとも部分的に囲むように構成される、請求項31に記載のカテーテル。
- 前記染料キャリアの一部分を所望の着目組織の一部分に接触させて位置付けるための前記手段は、前記所望の着目組織に対して前記染料キャリアを位置付けるように、前記被検体内で前記カテーテルシースを導くための手段を含む、請求項32に記載のカテーテル。
- 前記光エネルギー源は、レーザーである、請求項21に記載のカテーテル。
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US8620409B2 (en) | 2008-08-04 | 2013-12-31 | University Of Utah Research Foundation | Dye application for confocal imaging of cellular microstructure |
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JP7060891B2 (ja) | 2016-05-18 | 2022-04-27 | 国立大学法人三重大学 | がん検査装置、がん検査方法、および、がん検査用の染色剤 |
WO2020202742A1 (ja) * | 2019-03-29 | 2020-10-08 | テルモ株式会社 | カテーテル組立体、発光方法およびカテーテル操作の訓練方法 |
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US8620409B2 (en) | 2013-12-31 |
CA2732962A1 (en) | 2010-02-11 |
EP2324362B1 (en) | 2017-12-13 |
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US20110301438A1 (en) | 2011-12-08 |
WO2010016885A1 (en) | 2010-02-11 |
EP2324362A4 (en) | 2014-02-26 |
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