Intracellular Monitoring of AS1411 Aptamer by Time-Resolved Microspectrofluorimetry and Fluorescence Imaging

• Published in: Journal of fluorescence Vol. 25; no. 5; pp. 1245 - 1250
• Main Authors: Kočišová, Eva, Praus, Petr, Bok, Jiří, Bonneau, Stéphanie, Sureau, Franck
• Format: Journal Article
• Language: English
• Published: New York Springer US 01.09.2015; Springer Verlag
• Subjects: Analytical Chemistry; Aptamers, Nucleotide - metabolism; Base Sequence; Biochemistry; Bioengineering; Biological and Medical Physics; Biological Physics; Biomedical and Life Sciences; Biomedicine; Biophysics; Biotechnology; Cell Line, Tumor; Humans; Imaging; Intracellular Space - genetics; Intracellular Space - metabolism; Life Sciences; Microscopy, Fluorescence - methods; Oligodeoxyribonucleotides - genetics; Oligodeoxyribonucleotides - metabolism; Original Article; Physics; Spectrometry, Fluorescence - methods; Time Factors; Fluorescence microscopy imaging; Aptamer; AS1411 aptamer; Phase-modulation lifetime measurement; Intracellular fluorescence; intracellular fluorescence; phase modulation fluorescence lifetime 2; aptamer; micro imaging
• ISSN: 1053-0509; 1573-4994
• DOI: 10.1007/s10895-015-1612-3

Time-resolved microspectrofluorimetry and fluorescence microscopy imaging—two complementary fluorescence techniques—provide important information about the intracellular distribution, level of uptake and binding/interactions inside living cell of the labeled molecule of interest. They were employed to monitor the “fate” of AS1411 aptamer labeled by ATTO 425 in human living cells. Confocal microspectrofluorimeter adapted for time-resolved intracellular fluorescence measurements by using a phase-modulation principle with homodyne data acquisition was employed to obtain emission spectra and to determine fluorescence lifetimes in U-87 MG tumor brain cells and Hs68 non–tumor foreskin cells. Acquired spectra from both the intracellular space and the reference solutions were treated to observe the aptamer localization and its interaction with biological structures inside the living cell. The emission spectra and the maximum emission wavelengths coming from the cells are practically identical, however significant lifetime lengthening was observed for tumor cell line in comparison to non–tumor one.