Abstract
We propose a conclusive difference observed between the excitation conditions required to observe porphyrins and copper-metallothioneins in cells and/or tissues using an ordinary fluorescence microscope. We have emphasized the importance of examining the spectral properties of the emissions to avoid any serious mistakes such as confusing porphyrins with copper-metallothioneins in the liver and kidneys. However, microspectrophotometry is not a conventional method for either histochemical, cytochemical, or pathological studies because microspectrophotometers are both expensive and difficult to operate. Therefore, we demonstrate a simple comparative method using ordinary excitation filter arrangements. When using our technique, it becomes possible to optically discriminate more accurately between the autofluorescence properties arising from porphyrins and those arising from copper-metallothioneins. We would like to name our simple technique “Triple Observation Method (TOM)”.
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Acknowledgements
This study was supported by grants-in-aid for Scientific Research from the Japan Society for the Promotion of Science (19590658). We are indebted to Drs. M. Kubota (Department of Pediatrics, Hokkaido University Graduate School of Medicine), K. Jin (Hokkaido Institute of Public Heath) and T. Okui (School of Veterinary Medicine, Rakuno Gakuen University) for their support during this study. We are grateful to our colleagues, especially Mrs. F. Takenaka and H. Mikami for their helpful animal management.
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Nakayama, K., Katoh, Y. & Tamura, M. “Triple Observation Method (TOM)” to Discriminate Optically Autofluorescence from Porphyrins Versus that from Copper-Metallothioneins. J Fluoresc 21, 835–839 (2011). https://doi.org/10.1007/s10895-010-0773-3
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DOI: https://doi.org/10.1007/s10895-010-0773-3