Health & Environmental Research Online (HERO)


Print Feedback Export to File
3038677 
Journal Article 
Selective labeling of lipid droplets in aldehyde fixed cell monolayers by lipophilic fluorochromes 
Stockert, JC; Abasolo, MI; Blazquez-Castro, A; Horobin, RW; Revilla, M; Lombardo, DM 
2010 
1 
Biotechnic & Histochemistry
ISSN: 1052-0295
EISSN: 1473-7760 
85 
5 
277-283 
English 
20560872 
WOS:000284272900001 
We evaluated a number of lipophilic dyes and fluorochromes, including oxazone and thiazone derivatives of oxazine and thiazine dyes, scintillator agents, a carotenoid and a metal-porphyrin complex for visualization of lipid droplets within aldehyde fixed cultured HeLa and BGC-1 cells. Observation under ultraviolet, blue or green exciting light revealed selective fluorescence of lipid droplets, particularly after treatment with aqueous solutions of Nile blue and brilliant cresyl blue oxazones, toluidine blue thiazone, or propylene glycol solutions of canthaxanthin, ethyl-BAO, and ZnTPyP. Mounting in water was required to maintain the fluorescence of lipids; the use of glycerol, Mowiol or Vectashield was not adequate. The effect of dye structure on staining intensity was assessed with the aid of numerical structure parameters modeling lipophilicity (HI and log P), overall size (MW) and the size of the conjugated system (conjugated bond number; CBN). The best stains for lipid droplets were relatively lipophilic (HI > 4.0, log P > 5.0), of small size overall (MW < 370), with small conjugated systems (CBN < 24), and not significantly amphiphilic. The two hydrophobic-hydrophilic parameters (the classic log P and the hydrophobic index, HI; values calculated by molecular modeling software) were highly correlated; however, HI was a more suitable hydrophobicity index for the dyes studied here. 
BBOT; brilliant cresyl blue oxazone; canthaxanthin; dm-POPOP; ethyl-BAO; fluorescent labels; lipid droplets; lipophilic dyes; log P; methylene blue thiazone; Nile red; oil red O; propylene glycol; Sudan III; Sudan IV; thionine thiazone; toluidine blue thiazone