Fine structural detection of calcium ions by photoconversion

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 combi...

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Bibliographic Details
Published in:European journal of histochemistry 2016-08, Vol.60 (3), p.2695-2695
Main Authors: Poletto, V, Galimberti, V, Guerra, G, Rosti, V, Moccia, F, Biggiogera, M
Format: Article
Language:English
Subjects:
Calcium - analysis
Calcium - metabolism
calcium ions
Calcium Signaling
Fluorescent Dyes - chemistry
Fura-2 - analogs & derivatives
Fura-2 - chemistry
HeLa Cells
Humans
Mag-Fura 2
Microscopy, Fluorescence - methods
Original Paper
photoconversion
Ultrastructural cytochemistry
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Summary: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.
ISSN:1121-760X
2038-8306
DOI:10.4081/ejh.2016.2695