JP7420950B2 - 多重蛍光インシトゥハイブリダイゼーション画像を獲得し、処理するためのシステム及び方法 - Google Patents
多重蛍光インシトゥハイブリダイゼーション画像を獲得し、処理するためのシステム及び方法 Download PDFInfo
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
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Description
for h = 1:N_hybridization
% multiple hybridizations
for f = 1:N_FOVs
% multiple lateral field-of-views
for c = 1:N_channels
% multiple color channels
for z = 1:N_planes
% multiple z planes
Acquire image(h, f, c, z);
end % end for z
end % end for c
end % end for f
end % end for h
例えば、
であり、ここで、iは、位置決め済みの画像310のスタック312(図3参照)におけるレイヤ番号を表わす。閾値は、ログのヒストグラムP(x,y)に基づいて生成されうる。
ここで、Ip,xはピクセルワードのマトリクス702からの値であり、Ci,xは参照コードワードのマトリクス722からの値である。ユークリッド距離の代わりに、その他のメトリック(例えば差の絶対値の和、余弦角、相関など)が使用されることもある。
BB=[I(p,3)+I(p,4)+I(p,5)+I(p,6)+I(p,12)+I(p,13)+I(p,14)]/7
Claims (12)
- 蛍光インシトゥハイブリダイゼーションイメージングシステムであって、
試薬中の蛍光プローブに曝露されるべきサンプルを包含するためのフローセルと、
複数の試薬源のうちの1つから前記フローセルへの流れを制御するためのバルブと、
前記フローセルを通るように流体を流すためのポンプと、
可変周波数励起光源及び前記サンプルから蛍光放出された光を受信するよう位置付けられたカメラを含む、蛍光顕微鏡と、
前記フローセルと前記蛍光顕微鏡とを垂直方向に相対運動させるためのアクチュエータと、
前記フローセルと前記蛍光顕微鏡とを横方向に相対運動させるためのモータと、
制御システムと、を備え、前記制御システムが、ネストされたループとして、
前記サンプルを複数の異なる試薬に曝露するために、前記バルブに、前記フローセルを複数の異なる試薬源に順次連結させることと、
前記複数の異なる試薬の各試薬につき、前記モータに、サンプルに対して、複数の異なる視野に前記蛍光顕微鏡を順次位置付けさせることと、
前記複数の異なる視野の各視野につき、前記可変周波数励起光源に、複数の異なる波長を順次放出させることと、
前記複数の異なる波長の各波長につき、前記アクチュエータに、サンプルに対して、複数の異なる垂直高さに前記蛍光顕微鏡を順次位置付けさせることと、
前記複数の異なる垂直高さの各垂直高さにつき、それぞれの波長によって励起されたそれぞれの試薬のそれぞれの蛍光プローブを有するサンプルのそれぞれの視野をカバーするそれぞれの垂直高さで、画像を取得することと、を行うよう構成される、
蛍光インシトゥハイブリダイゼーションイメージングシステム。 - 前記可変周波数励起光源が複数のレーザモジュールを含み、前記制御システムは、前記複数の異なる波長を順次放出するために前記レーザモジュールを順次作動させるよう構成される、請求項1に記載のシステム。
- 前記制御システムが、前記バルブ及び前記モータを制御するためのコンピュータと、前記レーザモジュール及び前記アクチュエータを制御するためのマイクロコントローラとを含む、請求項2に記載のシステム。
- 前記サンプルと前記カメラとの間の光路内に位置付けられたフィルタホイールを更に備え、前記フィルタホイールは複数の発光バンドパスフィルタを含み、前記制御システムは、前記画像が取得される時に、選択されたフィルタが、前記複数の異なる波長のうちの前記波長によって励起された時の前記蛍光プローブによる発光に関連付けられた発光バンドパスを有するように、前記可変周波数励起光源と前記フィルタホイールとを協調させるよう構成される、請求項2に記載のシステム。
- 前記フローセル内の前記サンプルを支持するためのステージを備え、前記モータが、前記ステージを直行する軸の対に沿って駆動するよう構成されたリニアアクチュエータの対を備える、請求項1に記載のシステム。
- 前記複数の異なる視野のうちの少なくとも2つの視野の各視野、及び前記視野の複数の画像において共有される少なくとも2つのピクセルの各ピクセルにつき、コードブック内の複数のコードワードから、前記視野での前記複数の画像における前記ピクセルについてのデータ値との最良一致を提供するコードワードを特定することによって、前記ピクセルを復号するよう構成された、データ処理システムを更に備える、請求項1に記載のシステム。
- 各画像につき画質値を決定し、前記画質値が画質閾値を満たさなければ画像を選別排除するよう構成されるデータ処理システムをさらに備える、請求項1に記載のシステム。
- 品質メトリックは、鮮鋭度品質値であり、品質メトリック閾値は、鮮鋭度品質閾値である、請求項7に記載のシステム。
- サンプルを包含するためのフローセルと、前記サンプルを照射するための可変周波数励起光源と、蛍光顕微鏡とを含む蛍光インシトゥハイブリダイゼーションイメージングシステムの制御のためのコンピュータプログラム製品であって、命令を有するコンピュータ可読媒体を備え、前記命令が、一又は複数のプロセッサに、
ネストされたループとして、
前記サンプルを複数の異なる試薬に曝露するために、バルブに、前記フローセルを複数の異なる試薬源に順次連結させることと、
前記複数の異なる試薬の各試薬につき、モータに、サンプルに対して、複数の異なる視野に前記蛍光顕微鏡を順次位置付けさせることと、
前記複数の異なる視野の各視野につき、前記可変周波数励起光源に、複数の異なる波長を順次放出させることと、
前記複数の異なる波長の各波長につき、アクチュエータに、サンプルに対して、複数の異なる垂直高さに前記蛍光顕微鏡を順次位置付けさせることと、
前記複数の異なる垂直高さの各垂直高さにつき、それぞれの波長によって励起されたそれぞれの試薬のそれぞれの蛍光プローブを有するサンプルのそれぞれの視野をカバーするそれぞれの垂直高さで、画像を取得することと、を実行させる
コンピュータプログラム製品。 - 前記複数の異なる視野のうちの少なくとも2つの視野の各視野、及び前記視野の複数の画像において共有される少なくとも2つのピクセルの各ピクセルにつき、コードブック内の複数のコードワードから、前記視野での前記複数の画像における前記ピクセルについてのデータ値との最良一致を提供するコードワードを特定することによって、前記ピクセルを復号する命令を更に備える、請求項9に記載のコンピュータプログラム製品。
- 各画像につき画質値を決定し、前記画質値が画質閾値を満たさなければ画像を選別排除する命令を更に備える、請求項9に記載のコンピュータプログラム製品。
- 品質メトリックは、鮮鋭度品質値であり、品質メトリック閾値は、鮮鋭度品質閾値である、請求項11に記載のコンピュータプログラム製品。
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TW202138564A (zh) | 2021-10-16 |
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