C2C12 myotubes were washed with PBS and fixed with 4% paraformaldehyde for 15 min. LipidTOX neutral lipid stain (Thermo Fisher Scientific) and DAPI were added. Cells are incubated 30 min at room temperature before imaging. Labeling patterns were observed using the Zeiss LSM 710 confocal microscope.
Lipidtox neutral lipid stain
Manufactured by Thermo Fisher Scientific
Sourced in United States
LipidTOX neutral lipid stain is a fluorescent dye used to detect and visualize neutral lipids in cells. It binds to neutral lipid droplets, allowing for their detection and quantification using fluorescence microscopy or flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 710 confocal microscope, by Zeiss (3 mentions)
Hoechst 33342, by Thermo Fisher Scientific (2 mentions)
Hoechst staining, by Thermo Fisher Scientific (2 mentions)
Lipidtox staining, by Thermo Fisher Scientific (2 mentions)
μclear, by Greiner (1 mentions)
Dm2500, by Leica (1 mentions)
Anti myosin heavy chain antibody, by Abcam (1 mentions)
Retiga q image digital still camera, by Nikon (1 mentions)
Act 1, by Nikon (1 mentions)
Confocal laser scanning microscopy, by Leica (1 mentions)
8 protocols using lipidtox neutral lipid stain
1
Visualizing Lipid Droplets in C2C12 Myotubes
2
Quantification of Lipid Accumulation
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Cells were plated in clear bottom 96-well plates (μClear, Greiner) and treated with FFAs or BSA as described in the legend of the figure, and after 12 h, the cells were fixed in 4% neutral-buffered formalin, for 30 min, at room temperature. The fixative was removed, and cells were washed twice and then added PBS solution containing LipidTOX Neutral Lipid Stain (Thermo Fisher Scientific). The cells were incubated for 30 min at room temperature, the staining solution removed, and a solution containing Hoechst 33342 (1 μg/ml) (Thermo Fisher Scientific) was added for 20 min, at room temperature. The solution was removed, and the cells were washed twice in PBS, and the fluorescence signal was acquired at 577/609 nm (Ex/Em) for LipidTOX and 350/461 nm (Ex/Em) for Hoechst 33342. The lipid accumulation was the ratio between the lipidTOX and Hoechst 33342 fluorescence and expressed as a percentage related to control.
3
Lipid Staining of Frozen Eyelid Sections
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Frozen eyelid sections were fixed in 4% paraformaldehyde for 10 minutes, rinsed in PBS for 5 minutes, diluted the LipidTOX neutral lipid stain (H34475; Thermofisher, Waltham, MA, USA) 1:200 in buffer and incubated the sections at room temperature for at least 30 minutes, followed by counterstaining with DAPI. Sections were then evaluated and imaged with a microscope (DM2500; Leica Microsystems, Wetzlar, Germany).
4
Urethral Muscle Lipid Staining Protocol
5
Visualizing Lipids in Skin Organoids
6
Lipid Modulation by Pannexin-1 and FAM3A
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Cells infected with Ad-PANX1 or Ad-FAM3A in the presence of free fatty acids (FFAs, 0.2 mmol/L oleic acid and 0.4 mmol/L palmitic acid) and PBN (200 μmol/L) or suramin (50 μmol/L) for 24 h (Ad-GFP as control treatment). Cells were fixed with 4% paraformaldehyde for 15 min, then stained with LipidTOX™ neutral lipid stain (1:1000; #H34476, Invitrogen, USA). After nuclear staining with DAPI, the cells on coverslips were visualized and quantified with Leica Confocal Laser Scanning Microscopy.
7
Lipid Quantification in FFA-Treated Hepatocytes
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HepG2 cells plated on coverslips were treated with FFAs (0.1 mmol/L oleic acid and 0.2 mmol/L palmitic acid) in the absence or presence of drugs. Cells were washed with PBS twice after 24-h treatment and fixed with 4% paraformaldehyde for 10 min. We diluted the LipidTOX neutral lipid stain (Invitrogen) 1:1,000 in buffer to make a 1× solution. For every coverslip, 100 μL solution is required for 30 min at room temperature. After nuclear staining with DAPI, coverslips were mounted on glass slides using 50% glycerol in PBS. Mounted coverslips were imaged and cells were visualized by fluorescence microscopy using a confocal laser scanning microscope. For quantitative determination of triglyceride (TG) content in cells, treated hepatocytes were lysed in lysis buffer (TG determination kit, cat. no. E1013; Applygen Technologies, Beijing, China), and TG content was measured according to the manual of the kit. The TG content (μmol) was normalized to the protein content in the same sample (μmol/mg protein).
8
Visualizing Lipids in Skin Organoids
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LipidTOX Staining (Invitrogen) was performed to visualize sebaceous glands and lipid-rich adipocytes. Cryosections of skin organoids were incubated with LipidTOX neutral lipid stain diluted at a ratio of 1:200 in 1X PBS for 30 min at RT, followed by Hoechst staining (1:2000; Invitrogen) for 1 min at RT to visualize nuclei.
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