METHODOLOGICAL APPROACHES TO EVALUATION OF INTRACELLULAR DISTRIBUTION OF LIPID AND AQUEOUS PHASE OF LIPOSOMES USING FLUORESCENT PROBES
Rinat A. Mukhamadiyarov, Igor G. Khaliulin, Evgeniya A. Sergeeva, Anna C. Tikhonova, Elena A. Velikanova, Olga D. Sidorova, Alexey S. Golovkin
Pages: 218-227 Published: 2 Jun 2014
Views: 2,259 Downloads: 1,123
Abstract: Creation of targeted liposomal delivery systems of biologically active substances to the tissues, cells and organelles is one of the topical problems of cellular medicine. These studies require solving the problem of detection of the precise localization of substances delivered with liposomes. Complexity of this problem is associated with invisibility of liposomes in the light microscope because of the small size, lack of colour and autoluminescence. A possible way of solving this problem is to visualize the liposomes by including in their composition fluorescent dyes. In the study with fluorescent or confocal microscope, colocalization of liposomal dyes and dyes for differential staining of intracellular organelles may serve as evidence of the presence of liposomes in this compartment. For these studies, an important prerequisite is an adequate choice of dyes for labeling liposomes and organelles. In this work, we investigate the methodological approaches of the study of intracellular localization of the liposomes in the cells using vital dyes for labeling liposomes and intracellular organelles. On specific examples, we demonstrate that the technology of the assessment of intracellular distribution of fluorescence using labeling of hydrophilic and hydrophobic parts of the liposomes and determination of the colocalization of these labels and organelle-specific fluoresce is effective and can be used in experiments aiming to create liposomal formulations for targeted drug delivery.