Visualizing endocytotic pathways at transmission electron microscopy via diaminobenzidine photo-oxidation by a fluorescent cell-membrane dye

Abstract

The endocytotic pathway involves a complex, dynamic and interacting system of intracellular compartments. PKH26 is a fluorescent dye specific for long-lasting cell membrane labelling which has been successfully used for investigating cell internalization processes, at either flow cytometry or fluorescence microscopy. In the present work, diaminobenzidine photo-oxidation was tested as a procedure to detect PKH26 dye at transmission electron microscopy. Our results demonstrated that DAB-photo-oxidation is a suitable technique to specifically visualise this fluorescent dye at the ultrastructural level: the distribution of the granular dark reaction product perfectly matches the pattern of the fluorescence staining, and the electron density of the fine precipitates makes the signal evident and precisely detectable on the different subcellular compartments involved in the plasma membrane internalization routes.

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Author Biographies

S. Grecchi, University of Verona
Department of Neurological and Movement Sciences, Anatomy and Histology Section
M. Malatesta, University of Verona
Department of Neurological and Movement Sciences, Anatomy and Histology Section
Published
2014-10-22
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Section
Technical Notes
Keywords:
Endocytosis, diaminobenzidine photo-oxidation, transmission electron microscopy.
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How to Cite
Grecchi, S., & Malatesta, M. (2014). Visualizing endocytotic pathways at transmission electron microscopy via diaminobenzidine photo-oxidation by a fluorescent cell-membrane dye. European Journal of Histochemistry, 58(4). https://doi.org/10.4081/ejh.2014.2449