Development of an aerosol flow cytometry technology for online measurement of airborne microorganisms

• Project/Area Number: 16H02934
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Environmental dynamic analysis
• Research Institution: Kanazawa University
• Principal Investigator: Furuuchi Masami 金沢大学, 地球社会基盤学系, 教授 (70165463)
• Co-Investigator(Kenkyū-buntansha): 畑 光彦 金沢大学, 地球社会基盤学系, 准教授 (00334756) ; 瀬戸 章文 金沢大学, フロンティア工学系, 教授 (40344155) ; 那須 正夫 大阪大谷大学, 薬学部, 客員教授 (90218040)
• Research Collaborator: SUTHIDA PIRIYAKARNSAKUL ; MUHAMMAD AMIN
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥18,200,000 (Direct Cost: ¥14,000,000、Indirect Cost: ¥4,200,000); Fiscal Year 2018: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2017: ¥7,930,000 (Direct Cost: ¥6,100,000、Indirect Cost: ¥1,830,000); Fiscal Year 2016: ¥6,110,000 (Direct Cost: ¥4,700,000、Indirect Cost: ¥1,410,000)
• Keywords: バイオエアロゾル / 蛍光染色 / 環境計測 / 気中微生物 / 蛍光色素 / 染色時間 / 気中染色 / エレクトロスプレー

Outline of Final Research Achievements

A rapid method for the online detection of bio-aerosols could be useful for the efficient management of areas that are contaminated with bio-aerosols. Airborne microbes of interest can be stained by exposing them to a fluorescent dye in the form of a mist, and the resulting stained microbes can be detected by an optical detection method. To accomplish this, we have tested the possibility as follows:
1) The rapid staining of yeast (S. cerevisiae) in an aqueous solution of typical dyes, such as DAPI and Auramine-O, was examined using a fluorescence spectrophotometer. The relaxation time, which corresponds to ~63.2 % of the maximum fluorescence intensity, could be adjusted to below 10 seconds.
2) Mixtures of aerosols of microbes and a mist of a fluorescent dye solution were examined as a possible method for staining microbes. By simply mixing an aerosol yeast and dye mist, a certain fraction of yeast could be confirmed to be stained.

Academic Significance and Societal Importance of the Research Achievements

現在最も信頼ある微生物の迅速測定法は微生物を液中で蛍光染色し，観察またはレーザー散乱光から情報抽出する「フローサイトメトリー（FCM）」によるものであるが，「気中では蛍光染色できない」ため，気中微生物も液中へトラップした後，培養や蛍光染色を行う必要があり，この時間と手間が迅速性を損なっていた。本研究では「気中微生物蛍光染色技術」に必須となる「短時間（１０秒以内）での蛍光染色」「気中での微生物粒子と蛍光染料の直接混合による染色」の可能性を検証し，世界で初めての「気中微生物の浮遊状態での蛍光染色」の成功例を示した。本成果は今後気中微生物の「迅速で正確なその場測定技術」の開発へ応用される予定である。

Research Products

• [Journal Article] Estimation of air pollution from ribbed smoked sheet rubber in Thailand exports to Japan as a pre-product of tires (2019)
  • Author(s): Phairuang Worradorn, Tekasakul Perapong, Hata Mitsuhiko, Tekasakul Surajit, Chomanee Jiraporn, Otani Yoshio, Furuuchi Masami
  • Journal Title: Atmospheric Pollution Research Volume: 10 Issue: 2 Pages: 642-650
  • DOI: 10.1016/j.apr.2018.11.010
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] The influence of the open burning of agricultural biomass and forest fires in Thailand on the carbonaceous components in size-fractionated particles (2019)
  • Author(s): Phairuang Worradorn, Suwattiga Panwadee, Chetiyanukornkul Thaneeya, Hongtieab Surapa, Limpaseni Wongpun, Ikemori Fumikazu, Hata Mitsuhiko, Furuuchi Masami
  • Journal Title: Environmental Pollution Volume: 247 Pages: 238-247
  • DOI: 10.1016/j.envpol.2019.01.001
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Polycyclic aromatic hydrocarbons and their nitro-derivatives from indoor biomass fueled cooking in two rural areas of Thailand: a case study. (2017)
  • Author(s): Orakij, W., Chetiyanukornkul, T., Kasahara, C., Boongla, Y., Chuesaard, T., Furuuchi, M., Hata, M., Tang, N., Hayakawa, K., Toriba, A.
  • Journal Title: Air Qual. Atmos. Health Volume: 印刷中 Issue: 6 Pages: 747-761
  • DOI: 10.1007/s11869-017-0467-y
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] An Investigation of the Time Dependency for the Staining Process of Microbes: the Behavior of Microbes in relation to Bio-aerosol Online Monitoring (2017)
  • Author(s): Suthida Piriyakarnsakul , Kaoru Takarada, Mitsuhiko Hata, Masami Furuuchi
  • Journal Title: 日本海域研究 Volume: 49 Pages: 1-10
  • NAID: 120006471377
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] オンライン計測を目的とした微生物気中染色法の検討 (2018)
  • Author(s): ピリヤカンサクン スティダー, 寶田 馨, 那須正夫, 畑 光彦, 古内正美
  • Organizer: 第35回エアロゾル科学・技術研究討論会
• [Presentation] Investigation into Time Dependency of Staining Process of Microbes for Bio-aerosol Online Monitoring (2017)
  • Author(s): Suthida Piriyakarnsakul , Kaoru Takarada, Mitsuhiko Hata, Masami Furuuchi
  • Organizer: 3rd International Symposium to Promote Joint Usage/Research Center “International Collaboration Research Base for Reaction of Atmosphere-Marine-Ecosystem Caused by Aerosol”
  • Int'l Joint Research