JP4758507B2 - 血小板の計数 - Google Patents
血小板の計数 Download PDFInfo
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- JP4758507B2 JP4758507B2 JP2009516448A JP2009516448A JP4758507B2 JP 4758507 B2 JP4758507 B2 JP 4758507B2 JP 2009516448 A JP2009516448 A JP 2009516448A JP 2009516448 A JP2009516448 A JP 2009516448A JP 4758507 B2 JP4758507 B2 JP 4758507B2
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Description
Claims (35)
- 血液試料中の血小板の体積計数のための試料獲得装置であって、
血液試料を受け取るための測定空洞であって、所定の固定された厚さを有する測定空洞と、
該測定空洞を画定する表面上に乾燥した形で配置された試薬であって、該血液試料中の赤血球を溶解させるための溶血剤を含み、且つ該血液試料中の血小板を選択的に染色するための染色剤をさらに含む試薬とを含む、上記試料獲得装置。 - 前記試料獲得装置が、前記測定空洞を画定する2つの平面を有する本体部材を含む、請求項1に記載の試料獲得装置。
- 前記平面が、光学測定のための試料の厚さを決定するように、互いから所定の距離のところに配置される、請求項2に記載の試料獲得装置。
- 前記測定空洞が、50〜170マイクロメートルの均一な厚さを有する、請求項1〜3までのいずれか一項に記載の試料獲得装置。
- 前記測定空洞が、80〜150マイクロメートルの均一な厚さを有する、請求項1〜3までのいずれか一項に記載の試料獲得装置。
- 前記染色剤が、前記血小板の膜を選択的に染色するように構成される、請求項1〜5までのいずれか一項に記載の試料獲得装置。
- 前記染色剤が、メチレンブルー、エオシンメチレンブルー、アズールエオシンメチレンブルー、Plaxan(商標)、ヘマトキシリン、メチレングリーン、トルイジンブルー、ゲンチアナバイオレット、ズダン類似体、ガロシアニン、又はフクシン類似体からなる群の中のいずれか1つである、請求項1〜6までのいずれか一項に記載の試料獲得装置。
- 前記溶血剤が、第4級アンモニウム塩、サポニン、胆汁酸、ジギトキシン、ヘビ毒、グルコピラノシド、又はトリトン(Triton)型の非イオン性界面活性剤である、請求項1〜7までのいずれか一項に記載の試料獲得装置。
- 前記溶血剤が、サポニンである、請求項1〜7までのいずれか一項に記載の試料獲得装置。
- 前記測定空洞を前記試料獲得装置の外部と連通させる試料入口をさらに含み、該入口が、血液試料を獲得するように構成される、請求項1〜9までのいずれか一項に記載の試料獲得装置。
- 血液試料中の血小板の体積計数のための方法であって、
血液試料を試料獲得装置の測定空洞内へ獲得するステップであって、該測定空洞が、溶血剤を含む試薬を保持し、該試薬が該測定空洞内に乾燥した形で配置されるステップと、
血小板を有する該試料に可視スペクトル全体の光を照射するステップと、
該測定空洞内の照射された試料の拡大像のデジタル画像を獲得するステップであって、位相差によって血小板を区別し、該デジタル画像が、少なくとも該測定空洞の厚さに相当する被写界深度で獲得されるステップと、
該デジタル画像をデジタル方式で分析して血小板を識別し、且つ該試料中の血小板の数を求めるステップとを含む、上記方法。 - 前記測定空洞において保持される試薬が、試料と反応して血小板を染色する染色剤を含み、染色剤で選択的に染色することによって且つ/又は位相差によって血小板を区別する、請求項11に記載の方法。
- 前記試料が、波長が前記染色剤の吸光度のピークに相当する光で照射される、請求項12に記載の方法。
- 前記血液試料が、前記測定空洞内で前記試薬と混合される、請求項11〜13のいずれか一項に記載の方法。
- 前記測定空洞が、50〜170マイクロメートルの厚さを有する、請求項11〜14のいずれか一項に記載の方法。
- 前記測定空洞が、80〜150マイクロメートルの厚さを有する、請求項11〜14のいずれか一項に記載の方法。
- 分析される試料の体積が、前記測定空洞の厚さ及び撮像される前記試料の面積によって明確に画定される、請求項15又は16に記載の方法。
- 前記照射するステップが、レーザ源を用いて実行される、請求項11〜17までのいずれか一項に記載の方法。
- 前記照射するステップが、発光ダイオードを用いて実行される、請求項11〜17までのいずれか一項に記載の方法。
- 前記デジタル画像が、3〜200倍の倍率を使用して獲得される、請求項11〜19までのいずれか一項に記載の方法。
- 前記デジタル画像が、4〜20倍の倍率を使用して獲得される、請求項11〜19までのいずれか一項に記載の方法。
- 前記分析するステップが、前記デジタル画像内の吸光度が高い領域を識別するステップを含む、請求項11〜21までのいずれか一項に記載の方法。
- 前記分析するステップが、前記デジタル画像内の黒い点を識別するステップを含む、請求項22に記載の方法。
- 前記分析するステップが、前記獲得したデジタル画像を電子的に拡大するステップを含む、請求項11〜23までのいずれか一項に記載の方法。
- 血液試料中の血小板の体積計数のためのシステムであって、
請求項1〜10までのいずれか一項に記載の試料獲得装置と測定装置とを含み、該測定装置が、
測定空洞内に血液試料を保持する試料獲得装置を受けるように構成された試料獲得装置ホルダと、
血液試料に照射するように構成された光源と、
撮像システムであって、拡大システムと、測定空洞内の照射された試料の拡大像のデジタル画像を獲得するためのデジタル画像獲得手段とを含み、染色剤で選択的に染色することによって且つ/又は位相差によって該デジタル画像内で血小板を区別する撮像システムと、
獲得した該デジタル画像を分析して血小板を識別し、該血液試料中の血小板の数を求めるように構成された画像分析器とを含む、上記システム。 - 前記拡大システムが、少なくとも前記試料獲得装置の測定空洞の厚さの被写界深度で構成される、請求項25に記載のシステム。
- 分析される試料の体積が、前記測定空洞の厚さ及び撮像される前記試料の面積によって明確に画定される、請求項25又は26に記載のシステム。
- 前記光源が、波長が染色剤の吸光度のピークに相当する光を照射するように構成される、請求項25〜27のいずれか一項に記載のシステム。
- 前記光源が、レーザ源を含む、請求項25〜28までのいずれか一項に記載のシステム。
- 前記光源が、発光ダイオードを含む、請求項25〜28までのいずれか一項に記載のシステム。
- 前記拡大システムが、3〜200倍の倍率を有する、請求項25〜30までのいずれか一項に記載のシステム。
- 前記拡大システムが、4〜20倍の倍率を有する、請求項25〜30までのいずれか一項に記載のシステム。
- 前記画像分析器が、前記デジタル画像内の吸光度が高い領域を識別するように構成される、請求項25〜32までのいずれか一項に記載のシステム。
- 前記画像分析器が、前記デジタル画像内の黒い点を識別するように構成される、請求項33に記載のシステム。
- 前記画像分析器が、前記獲得したデジタル画像を電子的に拡大するように構成される、請求項25〜34までのいずれか一項に記載のシステム。
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