Fine Structural Detection of Calcium Ions by Photoconversion
| Autor: | Marco Biggiogera, Germano Guerra, Vittorio Rosti, V. Galimberti, Valentina Poletto, Francesco Moccia |
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| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Materials science Histology Fura-2 Mag-Fura 2 Biophysics chemistry.chemical_element Calcium Photochemistry Ion 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Calcium ions Photoconversion Ultrastructural cytochemistry Cell Biology photoconversion Molecule Humans Irradiation Calcium Signaling lcsh:QH301-705.5 Fluorescent Dyes calcium ions Original Paper 030104 developmental biology Membrane lcsh:Biology (General) chemistry Microscopy Fluorescence Transmission electron microscopy 030220 oncology & carcinogenesis Excited state HeLa Cells |
| Zdroj: | European Journal of Histochemistry : EJH European Journal of Histochemistry, Vol 60, Iss 3 (2016) |
| ISSN: | 2038-8306 1121-760X |
| Popis: | We propose a tool for a rapid high-resolution detection of calcium ions which can be used in parallel with other techniques. We have applied a new approach by photo-oxidation of diaminobenzidine in presence of the emission of an excited fluorochrome specific for calcium detection. This method combines the selectivity of available fluorophores to the high spatial resolution offered by transmission electron microscopy to detect even fluorescing molecules even when present in low amounts in membrane-bounded organelles. We show in this paper that Mag-Fura 2 photoconversion via diaminobenzidine oxidation is an efficient way for localizing Ca2+ ions at EM level, is easily carried out and reproducible, and can be obtained on a good amount of cells, since the exposition in our conditions is not limited to the direct irradiation of the sample via an objective but obtained with a germicide lamp. The end product is sufficiently electron dense to be detected clearly when present in sufficient amount within a membrane boundary. |
| Databáze: | OpenAIRE |
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