JP2014230514A - 微生物の検査方法 - Google Patents
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Abstract
Description
バラスト水は、通常、搭載する海域と異なる海域に排出されるため、該バラスト水に含まれるプランクトンや細菌等の微生物を本来の生息地以外の海域に運び、生態系を破壊する等の問題を引き起こす虞がある。
これにより、検体の液体中の生物の染色工程、液体中の生物の濃縮工程、液体中の生物の情報取得の工程等をフロー方式で行えるため、各方式をバッチ方式で行う手法と比べて、ひとつの工程を終えた検体の一部が次の工程に進むまでの待機時間を大幅に短縮、または0とすることができ、待機時間での染色の状態の劣化を防ぐ意味で安定した生物の生死の情報を取得することができるといったメリットがある。
これにより、段階的なサイズごとの微生物の捕捉を実現でき、その結果、サイズごとの基準で規制された許容残存基準を満たしているかどうかを迅速に測定できるといったメリットがある。
これにより、極めて短時間で微生物が明るく発光し、バラスト水中の微生物の量を簡便かつ短時間で計測することができる、また、蛍光発光の厚み部分が薄くなるため、バックグラウンドと微生物の蛍光発光との光量差が極めて明確となり、微生物の蛍光発光の検出精度を向上することができる、といった作用・効果がある。
本発明の第1実施例にあっては、前記試料調製工程が、一定量の試料及び前記蛍光染色試薬を撹拌・混合する蛍光染色工程と、該蛍光染色工程後の溶液を一定時間静置する静置工程と、該静置工程後の溶液に蛍光発光しない液体で希釈する希釈工程と、を含むものである。
また、本発明の第2実施例にあっては、前記試料調製工程が、一定量の試料及び前記蛍光染色試薬を撹拌・混合する蛍光染色工程と、該蛍光染色工程後の溶液を一定時間静置する静置工程と、該静置工程後の溶液にpH調整剤を添加するpH調整工程と、を含むものである。
diacetate)などを使用することができる。カルセインAMは、植物性プランクトンに対して染色しやすい傾向がある一方、FDAは、動物性プランクトンに対して染色しやすい傾向がある。本発明にあっては、染色試薬として、FDAを使用するのが好ましい。このとき、FDAの濃度が1mMである場合、試料容器2には、前記試料1容量に対して1%容量の染色試薬を投入するのが好ましい。すなわち、試料を1ml(ミリリットル)採取した場合は、1mM濃度のFDAを0.01ml(ミリリットル=10μL)添加し、試料を5ml(ミリリットル)採取した場合は、1mM濃度のFDAを0.05ml(ミリリットル=50μL)添加し、試料を10ml(ミリリットル)採取した場合は、1mM濃度のFDAを0.1ml(ミリリットル=100μL)添加するとよい。その後、作業者は試料容器2に回転子などの撹拌手段によって試料溶液を混合・撹拌する(図1の蛍光染色工程)。
2 試料容器
3 操作部
4 LED光源
5 励起光用バンドパスフィルタ
6 蛍光用バンドパスフィルタ
7 光電子増倍管(PMT)
8 CPU基板
9 表示部
Claims (5)
- 試料中の微生物量を測定するための微生物の検査方法であって、
前記試料中の微生物量の測定は、試料及び蛍光染色試薬を撹拌・混合して試料を調製する試料調製工程と、前記試料に特定波長の励起光を照射して得られた蛍光発光の発光数から微生物量を算出する微生物量算出工程と、を含むものであり、
前記試料調製工程は、一定量の試料及び前記蛍光染色試薬を撹拌・混合する蛍光染色工程と、該蛍光染色工程後の溶液を一定時間静置する静置工程と、該静置工程後の溶液に蛍光発光しない液体で希釈する希釈工程と、を含むことを特徴とする微生物の検査方法。 - 前記蛍光染色工程は、試料容器に該試料容器全体の容積の1〜10%容量の試料を投入するとともに、前記試料容器に前記試料1容量に対して1%容量の蛍光染色試薬を投入して撹拌・混合してなる請求項1記載の微生物の検査方法。
- 前記蛍光染色試薬は、FDAを使用し、希釈前の試料中の濃度が0.01mMとなるように添加してなる請求項2記載の微生物の検査方法。
- 試料中の微生物量を測定するための微生物の検査方法であって、
前記試料中の微生物量の測定は、試料及び蛍光染色試薬を撹拌・混合して試料を調製する試料調製工程と、前記試料に特定波長の励起光を照射して得られた蛍光発光の発光数から微生物量を算出する微生物量算出工程と、を含むものであり、
前記試料調製工程は、一定量の試料及び前記蛍光染色試薬を撹拌・混合する蛍光染色工程と、該蛍光染色工程後の溶液を一定時間静置する静置工程と、該静置工程後の溶液にpH調整剤を添加するpH調整工程と、を含むことを特徴とする微生物の検査方法。 - 前記pH調整工程は、前記蛍光染色工程での溶液のpHが8.0であったときに、当該pH調整工程にてpHが6.0となるようにpH調整剤を添加してなる請求項4記載の微生物の検査方法。
Priority Applications (10)
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JP2013112978A JP6238046B2 (ja) | 2013-05-29 | 2013-05-29 | 微生物の検査方法 |
US14/894,492 US9856506B2 (en) | 2013-05-29 | 2014-05-16 | Method for examining microorganisms |
SG11201509777WA SG11201509777WA (en) | 2013-05-29 | 2014-05-16 | Microorganism testing method |
EP14803792.2A EP3006569B1 (en) | 2013-05-29 | 2014-05-16 | Microorganism testing method |
DK14803792.2T DK3006569T3 (da) | 2013-05-29 | 2014-05-16 | Fremgangsmåde til test af mikroorganismer |
CN201480029593.2A CN105431544B (zh) | 2013-05-29 | 2014-05-16 | 微生物的检查方法 |
KR1020157036787A KR20160011672A (ko) | 2013-05-29 | 2014-05-16 | 미생물의 검사 방법 |
PCT/JP2014/063113 WO2014192565A1 (ja) | 2013-05-29 | 2014-05-16 | 微生物の検査方法 |
AU2014272067A AU2014272067B2 (en) | 2013-05-29 | 2014-05-16 | Method for examining microorganisms |
TW103118688A TWI622650B (zh) | 2013-05-29 | 2014-05-28 | 微生物之檢查方法 |
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US (1) | US9856506B2 (ja) |
EP (1) | EP3006569B1 (ja) |
JP (1) | JP6238046B2 (ja) |
KR (1) | KR20160011672A (ja) |
CN (1) | CN105431544B (ja) |
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USD776290S1 (en) * | 2014-12-31 | 2017-01-10 | W.H.P.M. Bioresearch And Technology | Fecal occult blood detector |
US11408815B2 (en) * | 2017-04-28 | 2022-08-09 | Microwise Aps | Method and a device for quantifying living organisms and use of a device |
US11286208B2 (en) | 2018-08-21 | 2022-03-29 | General Electric Company | Systems and methods for thermally processing CMC components |
GB201813976D0 (en) * | 2018-08-28 | 2018-10-10 | Johnson Matthey Plc | Method of monitoring a fluid and use of a tracer for monitoring a fluid |
KR102130242B1 (ko) * | 2019-01-30 | 2020-07-03 | 한국해양대학교 산학협력단 | 염색 혼합시약을 이용한 처리된 선박 평형수 검사방법 |
JP7322593B2 (ja) * | 2019-08-23 | 2023-08-08 | 株式会社サタケ | 微生物の検査装置及びその方法 |
KR102415611B1 (ko) * | 2020-01-14 | 2022-07-06 | (주) 한일사이메드 | 시료측정시스템 |
CN113430107A (zh) * | 2021-06-25 | 2021-09-24 | 常州市金坛第一人民医院 | 一种模块化的用于微生物检测的前处理系统 |
Citations (4)
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JPH03244395A (ja) * | 1990-02-21 | 1991-10-31 | Mitsubishi Heavy Ind Ltd | 微生物生細胞の計数方法 |
JP2004187534A (ja) * | 2002-12-09 | 2004-07-08 | Nitto Denko Corp | 微生物または細胞の生理活性の評価方法およびそのためのキット |
JP2007135582A (ja) * | 2005-10-19 | 2007-06-07 | Jfe Engineering Kk | バラスト水中の微生物検出方法および装置 |
JP2008054509A (ja) * | 2006-08-29 | 2008-03-13 | Yamaguchi Univ | 生菌、死菌及び疑似生菌の存在割合判別方法 |
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EP0653492B1 (en) * | 1993-11-15 | 2000-09-27 | Canon Kabushiki Kaisha | Process for obtaining individual microorganisms, and applications of that process |
JP2009085898A (ja) | 2007-10-03 | 2009-04-23 | Hitachi Ltd | 生物検査装置 |
JP6201285B2 (ja) | 2012-08-24 | 2017-09-27 | 株式会社サタケ | 微生物の検査方法及びその装置 |
TWI619809B (zh) * | 2012-08-24 | 2018-04-01 | 佐竹股份有限公司 | 微生物之檢查方法及其裝置 |
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JPH03244395A (ja) * | 1990-02-21 | 1991-10-31 | Mitsubishi Heavy Ind Ltd | 微生物生細胞の計数方法 |
JP2004187534A (ja) * | 2002-12-09 | 2004-07-08 | Nitto Denko Corp | 微生物または細胞の生理活性の評価方法およびそのためのキット |
JP2007135582A (ja) * | 2005-10-19 | 2007-06-07 | Jfe Engineering Kk | バラスト水中の微生物検出方法および装置 |
JP2008054509A (ja) * | 2006-08-29 | 2008-03-13 | Yamaguchi Univ | 生菌、死菌及び疑似生菌の存在割合判別方法 |
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EP3006569A1 (en) | 2016-04-13 |
KR20160011672A (ko) | 2016-02-01 |
US20160122796A1 (en) | 2016-05-05 |
TW201534729A (zh) | 2015-09-16 |
JP6238046B2 (ja) | 2017-11-29 |
AU2014272067A1 (en) | 2016-01-07 |
TWI622650B (zh) | 2018-05-01 |
US9856506B2 (en) | 2018-01-02 |
CN105431544B (zh) | 2019-06-14 |
EP3006569B1 (en) | 2019-07-10 |
WO2014192565A1 (ja) | 2014-12-04 |
EP3006569A4 (en) | 2017-05-10 |
DK3006569T3 (da) | 2019-10-14 |
CN105431544A (zh) | 2016-03-23 |
AU2014272067B2 (en) | 2020-01-30 |
SG11201509777WA (en) | 2015-12-30 |
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