Abstract
Fluorescence microscopy (FM) has recently been applied to the detection of airborne asbestos fibers that can cause asbestosis, mesothelioma and lung cancer. In our previous studies, we discovered that the E. coli protein DksA specifically binds to the most commonly used type of asbestos, chrysotile. We also demonstrated that fluorescent-labeled DksA enabled far more specific and sensitive detection of airborne asbestos fibers than conventional phase contrast microscopy (PCM). However, the actual diameter of the thinnest asbestos fibers visualized under the FM platform was unclear, as their dimensions were below the resolution of optical microscopy. Here, we used correlative microscopy (scanning electron microscopy [SEM] in combination with FM) to measure the actual diameters of asbestos fibers visualized under the FM platform with fluorescent-labeled DksA as a probe. Our analysis revealed that FM offers sufficient sensitivity to detect chrysotile fibrils as thin as 30–35 nm. We therefore conclude that as an analytical method, FM has the potential to detect all countable asbestos fibers in air samples, thus approaching the sensitivity of SEM. By visualizing thin asbestos fibers at approximately tenfold lower magnifications, FM enables markedly more rapid counting of fibers than SEM. Thus, fluorescence microscopy represents an advanced analytical tool for asbestos detection and monitoring.
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References
Mossman BT, Bignon J, Corn M, Seaton A, Gee JB (1990) Asbestos: scientific developments and implications for public policy. Science 247(4940):294–301
Perkins RL, Harvey BW (1993) Test method: Method for the determination of asbestos in bulk building materials. U. S. Environmental Protection Agency (EPA), Washington, DC
Mossman BT, Kamp DW, Weitzman SA (1996) Mechanisms of carcinogenesis and clinical features of asbestos-associated cancers. Cancer Invest 14(5):466–480
Theakston F (2000) In: Theakston F (ed) Air quality guidelines for Europe 2nd edn. World Health Organization, Regional Office for Europe, Copenhagen, pp 128–135
National Institute of Occupational Safety and Health (NIOSH) (1994) Asbestos and other fibers by PCM: Method 7400, in NIOSH Manual of Analytical Methods, 2nd issue, Washington, DC
National Institute of Occupational Safety and Health (NIOSH) (2010) Draft NIOSH Current Intelligence Bulletin: Asbestos Fibers and Other Elongate Mineral Particles: State of the Science and Roadmap for Research. NIOSH, Washington, DC
Stayner L, Kuempel E, Gilbert S, Hein M, Dement J (2008) An epidemiological study of the role of chrysotile asbestos fibre dimensions in determining respiratory disease risk in exposed workers. Occup Environ Med 65(9):613–619
Lippmann M (1988) Asbestos exposure indices. Environ Res 46(1):86–106
Steinmeyer R, Noskov A, Krasel C, Weber I, Dees C, Harms GS (2005) Improved fluorescent proteins for single-molecule research in molecular tracking and co-localization. J Fluoresc 15(5):707–721
Kuroda A, Nishimura T, Ishida T, Hirota R, Nomura K (2008) Detection of chrysotile asbestos by using a chrysotile-binding protein. Biotechnol Bioeng 99(2):285–289
Ishida T, Alexandrov M, Nishimura T, Minakawa K, Hirota R, Sekiguchi K, Kohyama N, Kuroda A (2010) Selective detection of airborne asbestos fibers using protein-based fluorescent probes. Environ Sci Technol 44(2):755–759
Robinson JM, Takizawa T, Pombo A, Cook PR (2001) Correlative fluorescence and electron microscopy on ultrathin cryosections: Bridging the resolution gap. J Histochem Cytochem 49(7):803–808
Atkinson AW, Gettings RB, Rickards AL (1970) Estimation of fibril lengths in chrysotile asbestos fibres. Nature 226(5249):937–938
Japanese Ministry of the Environment (2010) Asbestos Monitoring Manual, 4th rev, Tokyo (in Japanese), www.env.go.jp/air/asbestos/monitoring_manu/rev4_full.pdf
International Organization for Standardization (ISO) (2002) Ambient air - Determination of numerical concentration of inorganic fibrous particles—Scanning electron microscopy method (ISO 14966). ISO copyright office, Geneva
Kohyama N, Tanaka I, Tomita M, Kudo M, Shinohara Y (1997) Preparation and characteristics of standard reference samples of fibrous minerals for biological experiments. Ind Health 35(3):415–432
Acknowledgements
This work was supported by the Environment Research and Technology Development Fund (C-1101) of the Ministry of the Environment, Japan, and the Development of Systems and Technology for Advanced Measurement and Analysis Program of the Japan Science and Technology Agency. We thank Mr. Shigeaki Tachibana (SII Nanotechnology Inc.) and Mr. Kazunori Sugai and Mr. Shinji Iguchi (Carl Zeiss MicroImaging Co., Ltd) for technical assistance with correlative microscopic imaging.
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Takenori Ishida and Maxym Alexandrov contributed equally to this work.
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Ishida, T., Alexandrov, M., Nishimura, T. et al. Evaluation of Sensitivity of Fluorescence-Based Asbestos Detection by Correlative Microscopy. J Fluoresc 22, 357–363 (2012). https://doi.org/10.1007/s10895-011-0967-3
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DOI: https://doi.org/10.1007/s10895-011-0967-3