High resolution imaging of subcellular structures was performed in cells loaded with H2DCFDA expressing either endoplasmic reticulum-targeted or mitochondria-targeted red fluorescent protein (RFP) after a 15 minute incubation with 60 µM LPC 18:1 or PBS (vehicle). Images were acquired with an array confocal laser scanning microscope, built on an inverse, fully automatic microscope equipped with VoxCell Scan (VisiTech) and a 100× objective (Plan-Fluor 100×/1.45 Oil, Zeiss). H2DCFDA was illuminated at 488 nm (120 mW diode laser, Visitron Systems) and emission was collected at 535 nm (ET535/30 m, Chroma Technology Corp.). RFP was excited with 561 nm laser light (50 mW, VSLaserModul, Visitron Systems) and fluorescence was recorded at 630 nm (630/75, Chroma Technology Corp.). Emitted light was acquired with a CCD camera (CoolSNAP-HQ, Photometrics). Background correction and image overlay were performed using the MetaMorph 7.7.0.0 software.
Et535 30m
Manufactured by Chroma Technology
The ET535/30m is a bandpass filter designed for use in optical and imaging systems. It has a center wavelength of 535 nm and a bandwidth of 30 nm, allowing it to isolate a specific range of wavelengths from the visible light spectrum.
Automatically generated - may contain errors
Lab products found in correlation
Axiocam mrc, by Zeiss (2 mentions)
Plan fluor 100 1.45 oil, by Zeiss (1 mentions)
Observer z1, by Zeiss (1 mentions)
Axiovision software, by Zeiss (1 mentions)
Et500 20x, by Chroma Technology (1 mentions)
Phenol red, by Merck Group (1 mentions)
Fibronectin, by Merck Group (1 mentions)
M dish, by Ibidi (1 mentions)
Orca flash 4.0 camera, by Olympus (1 mentions)
Spectra x light engine, by Lumencor (1 mentions)
Lambda 10 3, by Sutter Instruments (1 mentions)
Dmi6000, by Leica (1 mentions)
3 protocols using et535 30m
1
Subcellular Imaging of LPC-Induced Oxidative Stress
2
Fluorescence Microscopy Imaging Protocol
3
Time-lapse and FRET Imaging Protocols
Check if the same lab product or an alternative is used in the 5 most similar protocols
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For phase-contrast time-lapse imaging assays, cells were seeded in 12-well tissue culture plates and imaged in CO 2 -independent L15 medium without phenol red (Life Technologies) supplemented with 10% FCS, at 203 using an inverted video-microscope (Olympus IX83) equipped with an ORCA flash 4.0 camera and controlled by CellSens software.
For FRET imaging assays, HeLa cells were seeded on glass-bottom dishes (m-Dish, IBIDI), precoated with fibronectin (Sigma-Aldrich) at 1 mg/cm 2 and imaged in L15 without phenol red supplemented with 1% FCS. Imaging was performed using an inverted microscope (Leica DMI6000) controlled by Metamorph software and equipped with adaptive focus control, fast emission filter wheel (lambda 10-3, Sutter), electron multiplying charge coupled device (EMCCD) camera (Evolve 512; Photometrics), HCX PL APO 403/NA 1.30 oil immersion lens, and light-emitting diode (LED)-based illumination system (spectra X-light engine, Lumencor). Filters used were ET480/40m and ET535/30m and double band CFP/YFP dichroic mirror 51017bs from Chroma (Chroma Technology). Quantifications were performed using ImageJ software (Gavet and Pines, 2010a) . Intensity-modulated display (IMD) representations were performed using Metamorph software.
For FRET imaging assays, HeLa cells were seeded on glass-bottom dishes (m-Dish, IBIDI), precoated with fibronectin (Sigma-Aldrich) at 1 mg/cm 2 and imaged in L15 without phenol red supplemented with 1% FCS. Imaging was performed using an inverted microscope (Leica DMI6000) controlled by Metamorph software and equipped with adaptive focus control, fast emission filter wheel (lambda 10-3, Sutter), electron multiplying charge coupled device (EMCCD) camera (Evolve 512; Photometrics), HCX PL APO 403/NA 1.30 oil immersion lens, and light-emitting diode (LED)-based illumination system (spectra X-light engine, Lumencor). Filters used were ET480/40m and ET535/30m and double band CFP/YFP dichroic mirror 51017bs from Chroma (Chroma Technology). Quantifications were performed using ImageJ software (Gavet and Pines, 2010a) . Intensity-modulated display (IMD) representations were performed using Metamorph software.
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