"Triple Observation Method (TOM)" to discriminate optically autofluorescence from porphyrins versus that from copper-metallothioneins.
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)".
Department of Health and Environmental Science, Hokkaido Institute of Public Health, North 19 West 12, Sapporo 060-0819 Hokkaido, Japan. ken_naka@iph.pref.hokkaido.jp
Kenji
K
Nakayama
Yoshinobu
Y
Katoh
Mamoru
M
Tamura
D000818:Animals; D002648:Child; D003116:Color; D003300:Copper / Q000737:chemistry*; D006527:Hepatolenticular Degeneration / Q000378:metabolism; D006801:Humans; D008099:Liver / Q000378:metabolism; D008297:Male; D008668:Metallothionein / Q000737:chemistry* / Q000378:metabolism*; D008856:Microscopy, Fluorescence / Q000379:methods*; D055620:Optical Phenomena; D011166:Porphyrins / Q000737:chemistry* / Q000378:metabolism*; D051381:Rats; D013050:Spectrometry, Fluorescence
D011166:Porphyrins; D003300:Copper; D008668:Metallothionein