Cells were seeded on 35mm glass bottom dishes (MatTek Corporation) and incubated with Liperfluo (10 μM) (Dojindo Molecular Technology, Inc.) or diaminorhodamine-4M (DAR-4AM) (5 μM) for 30 minutes at 37 °C. Cells were washed with Phosphate-Buffered Saline (PBS), the media replaced and the dish inserted into a closed, thermo-controlled (37ºC) stage top incubator (Tokai Hit Co., Shizuoka-ken, Japan) atop the motorized stage of an inverted Nikon TiE fluorescent microscope (Nikon Inc.) equipped with a 60X oil immersion optic (Nikon, CFI PlanFluor, NA 1.43) and NIS Elements Software. Liperfluo or DAR-4AM was excited using a Lumencor diode-pumped light engine (SpectraX, Lumencor Inc.) and detected using an ORCA-Flash 4.0 sCMOS camera (HAMAMATSU Corporation) and excitation and emission filters from Chroma Technology Corp. For time-lapse experiments, data was collected every 5 minutes for 3 hours, on approximately 10-20 cells per stage position, with 10-15 stage positions in each of 3 separate experiments per condition. Data were analyzed using NIS Elements (Nikon Inc).
Liperfluo
Manufactured by Lumencor
Liperfluo is a fluorometric detection system used to measure lipid content in cells and tissues. It provides a quantitative analysis of lipid levels through fluorescent labeling and detection.
Automatically generated - may contain errors
Lab products found in correlation
Nis element, by Nikon (2 mentions)
Liperfluo, by Dojindo Laboratories (2 mentions)
Spectra x, by Lumencor (2 mentions)
35 mm glass bottom dishes, by MatTek (1 mentions)
Tie fluorescent microscope, by Nikon (1 mentions)
Nis elements software, by Nikon (1 mentions)
Orca flash 4.0 scmos camera, by Hamamatsu Photonics (1 mentions)
Glass bottom tissue culture dishes, by MatTek (1 mentions)
Stage top incubator, by Tokai Hit (1 mentions)
2 protocols using liperfluo
1
Live-cell Fluorescent Imaging of Cellular Processes
2
Visualizing Lipid Peroxidation in Cells
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To visualize lipid peroxidation, cells were seeded in glass-bottom tissue culture dishes (35-mm, MatTek Corp) and prestained with MitoTracker® Deep Red FM (50 nM) and Liperfluo (10 μM, Dojindo Molecular Technologies Inc.,) for 30 min and then, RSL3 (0.5 μM), and/or Fer-1 (2 μM) were added, and the cells were incubated for an additional 1 h. The cells were washed twice with PBS, and then the dish was inserted into a closed, thermo-controlled (37 °C) stage top incubator (Tokai Hit Co.) above the motorized stage of an inverted Nikon TiE fluorescent microscope equipped with a 60x optic (Nikon, CFI Plan Fluor, NA 1.4). Liperfluo was excited using a diode-pumped light engine (SPECTRA X, Lumencor) and detected using an ORCA-Flash 4.0 sCMOS camera (Hamamatsu) and excitation and emission filters from Chroma. Data were collected on approximately 10–20 cells per stage position, with 10–15 stage positions per condition. Data was collected and analyzed using NIS Elements (Nikon Inc. Melville, NY).
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