Methods in Molecular Biology (2020) 1980: 47–53
DOI 10.1007/7651_2017_101
© Springer Science+Business Media New York 2017
Published online: 08 December 2017
Microalgae Lipid Staining with Fluorescent BODIPY Dye
Judita Koreiviene ˙
Abstract
The staining of lipids in algae cells with BODIPY dyes is much less studied compared to Nile red; therefore, a
complex of issues concerning staining details and fluorescence measurements still should be clarified for the
species that vary in cell wall complexity. Nevertheless, some general guidelines could be given, and a
preliminary protocol of the method is provided based on the existing data. The semiquantification of
lipid could be reliable if the staining protocol will be developed and adapted for particular microalgae
species.
Keywords Biofuel, Dye, Epifluorescence, Lipid staining, Oil, Screening
1 Introduction
Evaluation of the stained lipids in the cells by measuring fluorescence
intensity seems to be one of the most promising screening techniques. Nile red (NR) dye has been used most often to evaluate lipids in
microalgae [1–4]; however, the main disadvantages of these dyes are
that the accuracy and precision of staining differ depending on
strain/species of algae (cell wall structure and composition), growth
conditions, and development stage and that the dye is highly affected
by polarity of the solvent [3, 5]. As a potential alternative to NR, the
lipophilic fluorescent BODIPY dye has been recently used as a stain to
monitor algal oil storage within viable cells [3]. BODIPY dyes are very
promising for lipid determination due to their exceptional spectroscopic properties such as high molar extinction coefficient, sharp
emission bands, high fluorescent quantum yields, relative insensitivity to changes in pH and polarity, and good photochemical stability
[5–10]. BODIPY 505/515 was used to label a wide range of lipids such
as fatty acids, phospholipids, cholesterol, cholesteryl esters, and ceramides [11]. Compared to NR, BODIPY proved to have much better
and equal penetration through the different membranes and thus
more effectively staining lipids in microalgae that showed resistance
to photobleaching, maintaining its fluorescence longer than 30 min
[3, 5]. BODIPY 505/515 permeation is very fast due to its high
oil/water partition coefficient that allows dye to cross cell and organelle membranes easily [12, 13], and the attainment of a fluorescence
47
DOI 10.1007/7651_2017_101
© Springer Science+Business Media New York 2017
Published online: 08 December 2017
Microalgae Lipid Staining with Fluorescent BODIPY Dye
Judita Koreiviene ˙
Abstract
The staining of lipids in algae cells with BODIPY dyes is much less studied compared to Nile red; therefore, a
complex of issues concerning staining details and fluorescence measurements still should be clarified for the
species that vary in cell wall complexity. Nevertheless, some general guidelines could be given, and a
preliminary protocol of the method is provided based on the existing data. The semiquantification of
lipid could be reliable if the staining protocol will be developed and adapted for particular microalgae
species.
Keywords Biofuel, Dye, Epifluorescence, Lipid staining, Oil, Screening
1 Introduction
Evaluation of the stained lipids in the cells by measuring fluorescence
intensity seems to be one of the most promising screening techniques. Nile red (NR) dye has been used most often to evaluate lipids in
microalgae [1–4]; however, the main disadvantages of these dyes are
that the accuracy and precision of staining differ depending on
strain/species of algae (cell wall structure and composition), growth
conditions, and development stage and that the dye is highly affected
by polarity of the solvent [3, 5]. As a potential alternative to NR, the
lipophilic fluorescent BODIPY dye has been recently used as a stain to
monitor algal oil storage within viable cells [3]. BODIPY dyes are very
promising for lipid determination due to their exceptional spectroscopic properties such as high molar extinction coefficient, sharp
emission bands, high fluorescent quantum yields, relative insensitivity to changes in pH and polarity, and good photochemical stability
[5–10]. BODIPY 505/515 was used to label a wide range of lipids such
as fatty acids, phospholipids, cholesterol, cholesteryl esters, and ceramides [11]. Compared to NR, BODIPY proved to have much better
and equal penetration through the different membranes and thus
more effectively staining lipids in microalgae that showed resistance
to photobleaching, maintaining its fluorescence longer than 30 min
[3, 5]. BODIPY 505/515 permeation is very fast due to its high
oil/water partition coefficient that allows dye to cross cell and organelle membranes easily [12, 13], and the attainment of a fluorescence
47