JP7368539B2 - システムレベル較正 - Google Patents
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- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/36—Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements
- G02B21/365—Control or image processing arrangements for digital or video microscopes
-
- G—PHYSICS
- G06—COMPUTING; CALCULATING OR COUNTING
- G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
- G06T5/00—Image enhancement or restoration
- G06T5/90—Dynamic range modification of images or parts thereof
- G06T5/92—Dynamic range modification of images or parts thereof based on global image properties
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- H—ELECTRICITY
- H04—ELECTRIC COMMUNICATION TECHNIQUE
- H04N—PICTORIAL COMMUNICATION, e.g. TELEVISION
- H04N23/00—Cameras or camera modules comprising electronic image sensors; Control thereof
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- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- G01N2021/6439—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" with indicators, stains, dyes, tags, labels, marks
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- G—PHYSICS
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- G06T2207/00—Indexing scheme for image analysis or image enhancement
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- G06T2207/10064—Fluorescence image
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- G—PHYSICS
- G06—COMPUTING; CALCULATING OR COUNTING
- G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
- G06T2207/00—Indexing scheme for image analysis or image enhancement
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- G06T2207/10141—Special mode during image acquisition
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Description
本出願は、2017年6月28日に出願された米国仮特許出願第62/525,818号の出願日の利益を主張し、その開示は、参照によりその全体が本明細書に組み込まれる。
Claims (10)
- 1つまたは複数の染色剤で染色される生物試料を撮像する方法であって、各染色剤は前記生物試料内の特定の標的を識別し、当該方法は、
第1の所定のパワー値に達するように第1のチャネルについて光源の第1のパワー出力を設定すること、
カメラの第1の露出時間を調整することであって、それにより、前記第1のチャネルについて前記第1の所定のパワー値において第1の標的強度をもたらす、調整すること、
前記調整された第1の露出時間に基づいて、第1のチャネル内の前記生物試料の第1の画像を取得すること、および、
前記第1のチャネル内の前記1つまたは複数の染色剤の1つの染色剤に特異的な第1の導出画像補正係数に基づいて、取得した前記第1の画像内のピクセルデータを調整すること
を含み、
前記第1の導出画像補正係数は、(i)複数の染色剤について輝度値を正規化し、(ii)第1の染色剤について正規化された前記輝度値を整数1から減算することによって計算される、方法。 - 前記光源についての前記第1のパワー出力のレベルは、前記第1のチャネルについての既存の較正データから導出される、請求項1に記載の方法。
- 前記既存の較正データは、パワー出力対測定パワーの較正曲線である、請求項2に記載の方法。
- 前記第1の所定のパワー値は、特定のアッセイに特異的である、請求項1から3のいずれか一項に記載の方法。
- 前記第1の所定のパワー値は、全てのチャネルについて同じである、請求項4に記載の方法。
- 前記第1の標的強度は、前記第1のチャネルについての較正試料の標準化標的強度の所定のパーセンテージである、請求項1から5のいずれか一項に記載の方法。
- 前記所定のパーセンテージは、前記第1のチャネルについての前記標準化標的強度の約20%から約500%の範囲である、請求項6に記載の方法。
- 前記第1のチャネルについての前記標準化標的強度は、(i)複数の画像チャネルの各画像チャネルについての較正標準の較正強度値を決定し、(ii)前記複数の画像チャネルの各画像チャネルについての前記較正標準の前記較正強度値を、正規化較正強度値の各々の最大強度値に対して正規化し、(iii)前記複数の画像チャネルにわたって正規化強度の比を導出し、(iv)前記複数の画像チャネル内の最大輝度画像チャネルのために標的強度値を割り当て、(v)残りの画像チャネルについて標準強度値を計算することによって導出される、請求項6に記載の方法。
- 前記第1の画像内の前記ピクセルデータは、前記第1の画像内の各ピクセル値に第1の画像補正係数を乗算することによって調整される、請求項1から8のいずれか一項に記載の方法。
- 前記ピクセルデータを調整する前に、取得した前記第1の画像をアンミキシングすることをさらに含む、請求項1から9のいずれか一項に記載の方法。
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