Simple liquid-chromatographic method for Nile Red quantification in cell culture in spite of photobleaching.
Nile Red fluorescent marker is widely-used for different purposes, such as staining cell structures and for the visualization and localization of colloidal drug carriers. However, when fluorescence-dependent imaging or quantification is performed, the risk of inexact results is increased due to photobleaching. The proposed, simple quantification method of using an HPLC-UV-Vis system allows the determination of Nile Red even in photobleached samples. The intra- and inter-assay accuracies for all analytes were found to be within 94.9 and 100.8%, respectively, of target values. When samples underwent photobleaching by laser, UV-Vis detection varied at around 99+/-5%, whereas fluorescence decreased down to 86%. Such results show this method to be interesting for approaches where quantification should be performed after analysis such as fluorescent imaging.
Lamprecht
Alf
A
INSERM ERIT-M 0104, Université d'Angers, 10 rue André Boquel, 49100 Angers, France.
Benoit
Jean-Pierre
JP
D002138:Calibration; D018929:Cell Culture Techniques; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D010078:Oxazines / Q000032:analysis*; D010777:Photochemistry; D013056:Spectrophotometry, Ultraviolet
D010078:Oxazines; C044808:nile red