BODIPY 505/515

Procedure for BODIPY 505/515 staining of Aurantiochytrium sp. ^[1]

A 5 mM BODIPY 505/515 stock was prepared by dissolving in dimethyl sulfoxide and stored in the dark.

For efficient staining of the intracellular lipid bodies in Aurantiochytrium sp., an aliquot of 0.2% DMSO (vol/vol) was added to the microalgal suspension. 2 μL of BODIPY 505/515 solution was added into the l ml of algal-DMSO suspension and gently shaken for 1 min. Samples were then incubated in darkness for 5–10 min at room temperature. Then the samples were directly used for FACS and microscopic analyses

Procedure for BODIPY 505/515 staining of HepG2 and MCF7 cells. ^[2]

HepG2 and MCF7 cells were washed once with PBS and fixed in 4% paraformaldehyde for 15 min. Cells were washed three times with PBS and incubated with 0.2 μM BODIPY 505/515 for 30 min. The cells were then counterstained with 1.5 μg/mL DAPI for 5 min. Cells were examined and photographed using the fluorescence microscopy.

This protocol only provides a guideline, and should be modified according to your specific needs

References:

[1]. Velmurugan N, Sathishkumar Y, Yim S S, et al. Study of cellular development and intracellular lipid bodies accumulation in the thraustochytrid Aurantiochytrium sp. KRS101[J]. Bioresource technology, 2014, 161: 149-154.

[2]. Jhu J W, Yan J B, Lin Z H, et al. SREBP1-Induced Glutamine Synthetase Triggers a Feedforward Loop to Upregulate SREBP1 through Sp1 O-GlcNAcylation and Augments Lipid Droplet Formation in Cancer Cells[J]. International journal of molecular sciences, 2021, 22(18): 9814.