Bioimaging Nucleic-Acid Aptamers with Different Specificities in Human Glioblastoma Tissues Highlights Tumoral Heterogeneity.

Nucleic-acid aptamers are of strong interest for diagnosis and therapy. Compared with antibodies, they are smaller, stable upon variations in temperature, easy to modify, and have higher tissue-penetration abilities. However, they have been little described as detection probes in histology studies of human tissue sections. In this study, we performed fluorescence imaging with two aptamers targeting cell-surface receptors EGFR and integrin α5ß1, both involved in the aggressiveness of glioblastoma. The aptamers' cell-binding specificities were confirmed using confocal imaging. The affinities of aptamers for glioblastoma cells expressing these receptors were in the 100-300 nM range. The two aptamers were then used to detect EGFR and integrin α5ß1 in human glioblastoma tissues and compared with antibody labeling. Our aptafluorescence assays proved to be able to very easily reveal, in a one-step process, not only inter-tumoral glioblastoma heterogeneity (differences observed at the population level) but also intra-tumoral heterogeneity (differences among cells within individual tumors) when aptamers with different specificities were used simultaneously in multiplexing labeling experiments. The discussion also addresses the strengths and limitations of nucleic-acid aptamers for biomarker detection in histology.

Histofluorescence of monoamines in newt forelimb regenerates.

Newt forelimb regenerates were studied at various stages of development using the histofluorescent method of Falck and Hillarp. A green formaldehyde-induced fluorescence was found in nerve fibres, large dendritic cells, skin gland cells and skin gland cell secretions. To ascertain the nature of the fluorescent material, animals were submitted to treatments with L-dopa, nialamide, benserazide and reserpine, used separately or in combination and administered before cutting off the regenerates. The modifications of the fluorescence after the various treatments confirmed the monoaminic nature of the fluorophores. Catecholaminic fibres were numerous in tissues of fast-growing stages while in dedifferentiated cell areas as well as in prochondral cell condensations and in cartilage they were completely absent. Fluorescent dendritic cells that have never been described before in regenerating limbs were observed and, from their localisation and cytological appearance, classed as promelanophores (or melanoblasts).