JP4259879B2 - 身体領域内の標的の深さ、輝度およびサイズを決定するための方法および装置 - Google Patents
身体領域内の標的の深さ、輝度およびサイズを決定するための方法および装置 Download PDFInfo
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Description
本発明は、非侵襲性医学的画像化、医学的研究、病理学、ならびに薬物の発見および開発の分野のための装置および方法に関する。
種々の医学的診断および治療設定において、ならびに生体医学的研究において、被験体の身体領域内の表面下標的または領域を画像化することは望ましい。例えば、固形腫瘍の一部もしくは全て、心筋虚血の領域、被験体に投与された治療化合物の分布、または疾患の進行を非侵襲的に位置決定し、画像化することは、有用な研究または診断情報を提供し得る。理想的には、画像化方法は、身体領域内の目的の標的を位置決定し得、その身体領域の表面の下にあるその標的の形状、サイズ、細胞数、および深さについての情報を提供し得る。しかし、これまでは、表面下の身体標的画像化のために使用され、そして/または提唱されてきた方法は、一般に、X線のような電離放射線、磁気共鳴画像法(MRI)のような高価かつ大きくて扱いにくい機器、または超音波を使用する方法に制限されてきた。
1つの局面において、本発明は、被験体における発光源の位置、サイズおよび細胞数を調査するための方法を包含する。この方法を実施する際に、(i)発光源から発生し、(ii)被験体の混濁生物学的組織を通して進み、そして(iii)被験体の目的の第1の表面領域から発光する、光子を測定することによって構築される、第1の測定された光強度プロフィールを、光検出器デバイスを用いる第1の投射から、最初に得る。光強度プロフィールは、深さおよびサイズのような関数パラメータを予測するために、パラメータに基づく生物光学的関数に整合される。このように決定されたパラメータは、第1の測定された光強度プロフィール以外のデータを使用して精密化され、被験体における発光源のおよその深さおよび大きさを得る。さらなるデータは、被験体から測定されたデータ、モデリング分析からのデータ、または被験体の表面から放出された光子の波長に関連するデータであり得る。例えば、以下である:
この方法は、代表的に、被験体における発光源のおよその深さおよび形状を使用して、発光源の2−Dまたは3−Dの可視的表示を作成する工程、ならびにこの可視的表示を、被験体の2−Dまたは3−Dの画像に重ね合わせる工程を包含する。
(i)異なる波長で測定した相対光強度を決定し、そしてこの決定された相対光強度と、異なる波長における既知または算出された相対シグナル強度とを、組織深さの関数として比較すること;および
(ii)測定されたスペクトルを、組織内の種々の深さに配置された発光源から測定された複数のスペクトルと比較し、そしてこの測定されたスペクトルを既知のスペクトルと対応させることによって、発光源の深さを決定すること。
(1.定義)
他に明記されない限り、以下の用語は、以下のように規定される。
図1Aは、本発明に従って、被験体における発光源の位置およびサイズを調査するのに使用され得る装置20の断面図である。この装置は、一般的に、被験体からの発光現象が検出され得る遮光エンクロージャ22を備える。遮光エンクロージャ22は、共有に係るPCT公開番号WO200163247(これは、その全体において本明細書中に参考として援用される)に記載されるように構築され得る。簡単には、このチャンバは、後壁(例えば、後壁21)および側壁によって規定され、そして前開口部(図示されてないが、これは、開けられてチャンバの内側へのアクセスを提供し得、そして閉じられて遮光シールを提供し得る)を有する。遮光エンクロージャ22は、小動物に対して設計されたが、この技術は、ヒトを含むより大きな哺乳動物に適用され得る。
(a)第2投射角から被験体を投射することによって得られる、光強度データ、代表的には、空間プロフィールデータを意味する62で示されるような、第2投射データ。代表的には、装置内の被験体は、発光が、被験体表面の第2領域から得られるように光学システムに対して傾けられる;
(b)1つ以上のバンドパス範囲(例えば、青色光、緑色光、または赤色光のスペクトル範囲)で得られる光強度データを意味する64で示されるような、スペクトルデータ;
(c)代表的には、目的の被験体に近い被験体内の選択された位置で波長の関数として得られる予め決定された光強度値を意味する66で示されるような、透過率データ;
(d)組織パラメータ(例えば、光が拡散する被験体組織に対応する、減少した散乱係数μ’s、吸収係数μa、または有効係数μeff)の予め決定された値を意味する68で示されるような、組織特性データ。このデータは、例えば、拡散モデルにより作成される空間プロフィール曲線を精密化するために、使用される;
(e)被験体の標的領域および隣接表面を示すモデルにおけるか、またはこの被験体に近い実際の被験体において、選択された位置で既知の強度およびサイズの光源を配置することによって取得される、予め決定された光強度値、代表的には、空間プロフィールデータを意味する70で示されるような、シミュレーションデータ;ならびに
(f)空間プロフィール全体または検出器アレイ全体にわたって、合計または積分された全光強度を意味する72で示されるような、全強度データ。コンピュータデバイスにより実行されるプログラムは、60でさらなる情報を受信し、そして以下に考察されるように、74での発光源の深さおよびサイズを精密化するための情報を使用する。最終的に、発光源における、源の深さ、サイズ、ならびに必要に応じて、形状および細胞数の精密化された決定は、発光源の位置および強度を示すデータの形態および/または1つ以上の被験体画像のいずれかで利用者に表示される。
この節は、本発明を実施する際に収集され得る発光データの種々の型を考慮し、そしてこれらのデータが、モデル拡散曲線との曲線の整合に基づいて、光子拡散モデル化において、および/または発光源の深さもしくはサイズを決定するために、どのように使用され得るかを議論する。
この節は、身体の表面における、この表面の下の光源からの光子の放射、および混濁媒体を通る移動をシミュレートするための、光子伝播モデルを考慮する。特に、以下に基づいて、シミュレートされた光強度空間プロフィールを作成することが望ましい:(i)発光源の深さ、(ii)発光源のサイズ、ならびに(iii)光吸収係数および光散乱係数。光吸収係数および光散乱係数は、波長と、その光子が通って拡散する組織の性質との両方に依存するので、このモデルは、その組織の性質および選択された波長における空間プロフィールを、特に考慮するように精密化され得る。
D∇2φ(r)=μaφ(r)−S(r) (1)
であり、そして フィックの法則は、以下:
j(r)=−D∇φ(r) (2)
であり、ここで、φは、等方性の流束量(ワット/m2)であり、jは、小さい指向性流束(ワット/m2)であり、Sは、出力密度(ワット/m3)であり、rは、半径であり、そして
本発明の方法を実施する際に、被験体は、最初、上に詳述されたように、標的の源において、発光分子を局在化するために処理される。次いで、この被験体は、装置20(被験体のサイズに対して適切に縮尺を決められている)の遮光チャンバ内の選択された位置に配置され、そして光学システムが、発光源と検出オプティクスとの間の、被験体の表面領域における源からの発光事象を測定するように調節される。被験体を選択された光学的関係で固定した状態で、表面発光の光強度の空間プロフィールが得られ、これもまた上記のとおりである。図3Aは、表面下の光源から測定された光強度の表面マップであり、これを引き続いて使用して、図3Bに示される空間プロフィールを作成した。
合計または積分された光強度とは、上記のように、検出器アレイの規定された領域全体またはいくらかにわたって合計された光強度をいう。積分光強度は、式5の積分と比較されて、源の深さおよび輝度のなお別の評価を提供し得る。この情報は、プロフィール情報と組み合わせて提供され得る。積分光強度もまた、複数の波長について算出され得る。
(2波長空間画像を使用する、発光物体の深さの算出)
図13Aに示される空間情報を使用して、細胞の皮下注射を受けた動物(図13B)および肺に標識細胞を有する動物(図13C)におけるルシフェラーゼ標識細胞の深さを算出した。この分析を、以下の工程を使用して、600nmおよび640nmに等しい波長でのデータを使用して実施した:(i)生物発光細胞を、インビトロおよびインビボで、600nmおよび640nmで画像化した;(ii)これらの画像を、各画像についての平均強度の測定によって定量した;(iii)インビトロの画像データに対するインビボの画像データの比を、各波長において決定した;そして(iv)式7を使用して深さを算出し、そして図7における平均有効散乱係数μeffを、吸収係数および散乱係数から推定した。
Claims (8)
- 被験体における発光源の位置およびサイズを調査する際に使用される装置であって、該装置は、
遮光エンクロージャであって、該遮光エンクロージャの内部で該被験体からの発光事象が検出され得る、遮光エンクロージャと、
該エンクロージャ内に含まれる光学システムであって、該光学システムは、該エンクロージャ内の第1の投射から、光子を測定することによって構築される第1の光強度プロフィールを得る際に使用され、該光子は、(i)該発光源から生じ、(ii)該被験体の混濁生物学的組織を通じて進み、(iii)該被験体の目的の第1の表面領域から発せられる、光学システムと、
該光学システムに動作可能に接続された計算ユニットであって、(i)該第1の測定された光強度プロフィールをパラメータベースの生体光子関数に適合させ、(ii)該第1の測定された光強度プロフィールに加えてデータを用いて、該生体光子関数の該パラメータを精巧化することにより、該位置およびサイズの初期決定を改良して、該被験体における該発光源の精巧化された位置、深さ、形状、輝度を取得する計算ユニットと
を備える、装置。 - 前記光学システムは、冷却条件で動作され得る電荷結合デバイス(CCD)と、該CCD上に光を収束させるためのレンズとを含む、請求項1に記載の装置。
- 前記光学システムは、前記被験体の目的の複数の異なる表面領域から発せられた光子を検出するように設計されている、請求項2に記載の装置。
- 前記光学システムは、約400〜約800nmの間で選択された2以上の異なる波長範囲内で光子発光強度を測定する際に使用される、選択された異なる波長範囲内の光子を伝送する1以上の波長フィルタを含み、
前記計算ユニットは、
(i)該異なる波長にて測定された相対光強度を決定し、組織深さの関数として、該決定された相対光強度と、該異なる波長での既知の相対シグナル強度とを比較するというパラメータ精巧化動作、および
(ii)該測定されたスペクトルと、組織内の種々の深さに配置された発光源から測定される複数のスペクトルとを比較し、該測定されたスペクトルと、該既知のスペクトルとを整合させることから、該発光標的領域の深さを決定するというパラメータ精巧化動作
のうちの少なくとも一方を実行するように動作可能である、請求項1に記載の装置。 - 前記計算ユニットは、モデル被験体のモデル身体領域内の、モデル標的の発光のサイズ、形状、および/または深さの関数として、前記選択された波長範囲内で発光された光の空間分布に関する所定の空間分布情報を含むデータファイルを含み、該計算ユニットは、(i)前記第1の光強度プロフィールのスペクトル特性と、データベース中に含まれるスペクトル特性とを比較するためのタスクを実行する際に機能する、請求項1に記載の装置。
- 前記計算ユニットは、強度パターン画像を単一の強度値に積分し、特定のサイズおよび形状の発光源の関数として生成された、積分された光強度値により発光源サイズを推定するように動作可能である、請求項1に記載の装置。
- 前記計算ユニットは、生物学的組織の吸収特性および散乱特性に類似した吸収特性および散乱特性を有する混濁媒体を通した、複数の深さの1つに位置し、複数のサイズおよび複数の形状のうちの1つを有するグロー源からの光子拡散のモデルに基づく複数の理論的光強度プロフィールを含むデータベースを含み、該計算ユニットは、(i)該複数の理論的光強度プロフィールの各々と、第1の測定された光強度プロフィールとの間の適合の質を比較することと、(ii)最良の適合を該第1の測定された光強度プロフィールに提供する該理論的光強度プロフィールを選択することと、(iii)(ii)で選択された理論的光強度プロフィールからのパラメータを用いて、該被験体における該発光源のおよその深さおよび形状を取得することとを行うように動作可能である、請求項1に記載の装置。
- 前記計算ユニットは、前記被験体における発光源のおよその深さおよび形状を用いて、前記発光源の可視的な二次元表示または三次元表示を生成し、該可視的な表示を、該被験体の二次元画像または三次元画像に重ね合わせるように動作可能である、請求項1に記載の装置。
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US7616985B2 (en) * | 2002-07-16 | 2009-11-10 | Xenogen Corporation | Method and apparatus for 3-D imaging of internal light sources |
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DE60213993T2 (de) | 2007-03-15 |
US20030002028A1 (en) | 2003-01-02 |
US7403812B2 (en) | 2008-07-22 |
DE60213993D1 (de) | 2006-09-28 |
WO2002093143A3 (en) | 2004-01-08 |
CA2447262A1 (en) | 2002-11-21 |
US8825140B2 (en) | 2014-09-02 |
AU2002303819B2 (en) | 2007-03-01 |
US20100262019A1 (en) | 2010-10-14 |
JP2004528916A (ja) | 2004-09-24 |
KR20040012844A (ko) | 2004-02-11 |
ATE336717T1 (de) | 2006-09-15 |
US7764986B2 (en) | 2010-07-27 |
US8180435B2 (en) | 2012-05-15 |
EP1402243B1 (en) | 2006-08-16 |
EP1402243A2 (en) | 2004-03-31 |
US20070270697A1 (en) | 2007-11-22 |
US20120150026A1 (en) | 2012-06-14 |
WO2002093143A2 (en) | 2002-11-21 |
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