JP6811462B2 - 細胞塊に関する領域検出方法及び領域検出装置 - Google Patents
細胞塊に関する領域検出方法及び領域検出装置 Download PDFInfo
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Description
[1]領域検出装置の動作方法である領域検出方法であって、
細胞塊を撮像した画像を入力する入力ステップと、
前記入力ステップにおいて入力された画像から、前記細胞塊の輪郭を、当該輪郭に沿った順番と共に検出する輪郭検出ステップと、
前記輪郭検出ステップにおいて検出された輪郭及び順番に基づいて、前記細胞塊に含まれる特定の領域を検出する領域検出ステップと、
前記領域検出ステップにおいて検出された領域を示す情報を出力する出力ステップと、
を含む領域検出方法。
[2]前記輪郭検出ステップにおいて、前記画像に対して輪郭追跡処理を行って前記細胞塊の輪郭を検出する上記[1]に記載の領域検出方法。
[3]前記輪郭検出ステップにおいて、前記画像に対して前記輪郭追跡処理の前に平滑化処理を行う上記[2]に記載の領域検出方法。
[4]前記領域検出ステップにおいて、前記画像における前記輪郭から細胞塊に向かう方向の画素の輝度値に基づいて前記特定の領域を検出する上記[1]〜[3]の何れかに記載の領域検出方法。
[5]前記領域検出ステップにおいて、前記画像における前記輪郭の形状に基づいて前記特定の領域を検出する上記[1]〜[4]の何れかに記載の領域検出方法。
[6]前記領域検出ステップにおいて、前記画像における前記輪郭の形状が、前記輪郭を構成する各画素についての輪郭接線方向の曲率として表現され、当該曲率に基づき前記特定の領域を検出する上記[5]に記載の領域検出方法。
[7]前記特定の領域が、連続上皮構造の領域である上記[1]〜[6]の何れかに記載の領域検出方法。
[8]前記細胞塊が、網膜組織を含む上記[1]〜[7]の何れかに記載の領域検出方法。
[9]細胞塊を撮像した画像を入力する入力手段と、
前記入力手段によって入力された画像において、前記細胞塊の輪郭を、当該輪郭に沿った順番と共に検出する輪郭検出手段と、
前記輪郭検出手段によって検出された輪郭及び順番に基づいて、前記細胞塊に含まれる特定の領域を検出する領域検出手段と、
前記領域検出手段によって検出された領域を示す情報を出力する出力手段と、
を備える領域検出装置。
特徴1:細胞塊の外周に近い部分(表層)の輝度値が明るく(大きく)、細胞塊の表層から一定の深さの内部(細胞深層)の輝度値が小さいこと。
特徴2:細胞塊の輪郭がなめらかであること(即ち、曲率の変化量が少ないこと)。
特徴3:特徴1及び特徴2の形態上の特徴が、凝集体の接線方向に連続していること。
本特徴は、本発明者が見出し、本発明の基礎となる知見である。
(条件1)曲率の絶対値Cが、予め設定した閾値C1よりも小さい(例えば、C1は0.10)。
(条件2)「1/5」及び「2/5」の平均輝度値が、予め設定した閾値(例えば、700)よりも大きい。
(条件3)「3/5」及び「5/5」の平均輝度値が、予め設定した閾値(例えば、500)よりも小さい。
(条件4)当該点を含み、上記の条件1〜3を連続して満たす範囲(輪郭の長さ:L)が、予め設定した長さ(例えば、350)以上。
本実施例では、領域の検出対象となる細胞塊自体の生成についても説明する。ヒトiPS細胞(1231A3株、京都大学より入手)を「Scientific Reports,4,3594(2014)」に記載の方法に準じてフィーダーフリー培養した。フィーダーフリー培地としてはStemFit培地(AK03、味の素社製)、フィーダーフリー足場にはLaminin511-E8(ニッピ社製)を用いた。
特徴1.細胞塊の外周に近い部分(表層)の輝度値が明るく(大きく)、細胞塊の表層から一定深さの内部(細胞深層)の輝度値が暗い(小さい)こと。
特徴2.細胞塊の輪郭がなめらかであること(即ち、曲率の変化量が少ないこと)。
特徴3.特徴1及び特徴2の形態上の特徴が、凝集体の接線方向に連続していること。
なお、上述した条件1〜4は、上記の特徴1〜3の知見に基づいて得られたものである。
実施例1記載の方法で作製した浮遊培養開始後62日目のそれぞれ異なる3つの細胞塊を、実施例1記載の方法でカラーの明視野像(位相差像)を撮像した(図4〜図6(a))。この明視野像から実施例1記載の方法で目視で判定したところ、図4の画像の細胞塊が連続上皮構造を含んでいることを確認でき、一方、図5及び図6の細胞塊が連続上皮構造を含んでいないことがわかった。
実施例1記載の方法で作製した浮遊培養開始後63日目のそれぞれ異なる2つの細胞塊を、実施例1記載の方法でカラーの明視野像(位相差像)を撮像した(図7,図8(a))。
Claims (7)
- 領域検出装置の動作方法である領域検出方法であって、
細胞塊を撮像した画像を入力する入力ステップと、
前記入力ステップにおいて入力された画像から、前記細胞塊の輪郭を、当該輪郭に沿った順番と共に検出する輪郭検出ステップと、
(1)前記輪郭検出ステップにおいて検出された輪郭及び順番、並びに(2)前記画像における前記輪郭から細胞塊に向かう方向の画素の輝度値に基づいて、前記細胞塊に含まれる連続上皮構造の領域を検出する領域検出ステップと、
前記領域検出ステップにおいて検出された領域を示す情報を出力する出力ステップと、
を含む領域検出方法。 - 前記輪郭検出ステップにおいて、前記画像に対して輪郭追跡処理を行って前記細胞塊の輪郭を検出する請求項1に記載の領域検出方法。
- 前記輪郭検出ステップにおいて、前記画像に対して前記輪郭追跡処理の前に平滑化処理を行う請求項2に記載の領域検出方法。
- 前記領域検出ステップにおいて、前記画像における前記輪郭の形状に基づいて前記連続上皮構造の領域を検出する請求項1〜3の何れか一項に記載の領域検出方法。
- 前記領域検出ステップにおいて、前記画像における前記輪郭の形状が、前記輪郭を構成する各画素についての輪郭接線方向の曲率として表現され、当該曲率に基づき前記連続上皮構造の領域を検出する請求項4に記載の領域検出方法。
- 前記細胞塊が、網膜組織を含む請求項1〜5の何れか一項に記載の領域検出方法。
- 細胞塊を撮像した画像を入力する入力手段と、
前記入力手段によって入力された画像において、前記細胞塊の輪郭を、当該輪郭に沿った順番と共に検出する輪郭検出手段と、
(1)前記輪郭検出手段によって検出された輪郭及び順番、並びに(2)前記画像における前記輪郭から細胞塊に向かう方向の画素の輝度値に基づいて、前記細胞塊に含まれる連続上皮構造の領域を検出する領域検出手段と、
前記領域検出手段によって検出された領域を示す情報を出力する出力手段と、
を備える領域検出装置。
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