RAECs were cultured in 24‐well plate. Mitochondrial autophagosomes were detected according to the assay protocol (mitophagy detection kit, Dojindo, Japan). Mtphagy Dye accumulates in intact mitochondria, is immobilized on it with chemical bond and exhibits a weak fluorescence from the influence of surrounding condition. When mitophagy is induced, the damaged mitochondria fuse to lysosome and then, Mtphagy Dye emits a high fluorescence. After incubation of cells with 100 nmol/l Mtphagy Dye working solution at 37°C for 30 min., cells were treated with HG+Pal, HG+Pal+NaHS, HG+Pal+Mito‐TEMPO, HG+Pal+bafilomycin A1 and HG+ Pal+NaHS+bafilomycin A1 for 48 hrs. Bafilomycin A1 was used as control to identify mitochondrial autophagosome. Then, cells were incubated at 37°C for 30 min. with 1 μmol/l Lyso Dye working solution to observe the co‐localization of Mtphagy Dye and lysosome. The mitophagy phenomenon and the fusion of mitochondria with lysosome were observed by on a fluorescence microscope (Olympus, XSZ‐D2, Japan).
Xsz d2
Manufactured by Olympus
Sourced in Japan
The XSZ-D2 is a high-precision optical microscope designed for laboratory and research applications. It features a binocular viewing head with diopter adjustment and interpupillary distance control. The instrument is equipped with a LED illumination system and a range of objective lenses to provide clear, high-contrast images.
Automatically generated - may contain errors
Lab products found in correlation
C 7az, by Merck Group (6 mentions)
C 7az, by Olympus (3 mentions)
Mitophagy detection kit, by Dojindo Laboratories (2 mentions)
Mitotracker staining, by Beyotime (2 mentions)
Monodansylcadaverine mdc, by Solarbio (1 mentions)
Bodipy 558 568 c12, by Thermo Fisher Scientific (1 mentions)
Bodipy 493 503, by Thermo Fisher Scientific (1 mentions)
33 protocols using xsz d2
1
Mitophagy Induction and Lysosomal Fusion
2
Fluorescence Imaging of H2S in RAECs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The fluorescence intensity of H2S in RAECs was tested using 7‐azido‐4‐methylcoumarin (C‐7Az, Sigma‐Aldrich, USA), which has been proved to selectively respond to H2S. RAECs were incubated with 50 μM C‐7Az PBS for 30 min., followed by washing of the cells with PBS for three times. Visualization of the turn‐on fluorescence response of C‐7Az to H2S in RAECs was carried out using a fluorescence microscope (Olympus, XSZ‐D2, Japan). These results confirmed that excitation fluorescence imaging could be used to detect H2S through the triggered fluorescence response of C‐7Az.
3
Monodansylcadaverine-based Autophagy Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The A549 cells were seeded on coverslips overnight and, following the indicated irradiation treatments, 0.5 μM MDC was added to the cells, which were cultured for 1 h. The cells were then washed with phosphate-buffered saline (PBS) and fixed with a solution of 3.3% paraformaldehyde for 30 min. The coverslips were examined using fluorescence microscopy (Olympus XSZ-D2; Olympus Corporation, Tokyo, Japan).
4
Polysulfide Intracellular Monitoring using SSP4
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Intracellular production of polysulphide was monitored using a newly developed fluorescent probe, SSP4, with slight modifications.11 (link) Briefly, VSMCs were loaded with 50 μmol/L SSP4 in serum‐free DMEM containing 0.003% Cremophor EL for 15 minutes at 37℃ in the dark. After washing, SSP4 was detected using a fluorescence microscope (Olympus, XSZ‐D2).
5
Autophagic Vacuole Imaging with MDC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Monodansylcadaverine (MDC, Solarbio, China) has autofluorescence properties with an excitation wavelength at 365 nm, due to a dansyl group conjugated to cadaverine, a diamine-pentane. Under in vivo conditions, MDC accumulates as a selective fluorescent marker for autophagic vacuoles by interacting with membrane lipids that are highly concentrated in autophagic compartments. When MDC is incorporated into cells, the accumulation of this fluorescent reagent is observed in spherical compartments at the perinuclear region in spots distributed throughout the cytoplasm. Neonatal rat cardiomyocytes were incubated with 50 μM MDC in PBS at 37°C for 30 min. Autophagic vacuoles were analyzed using fluorescence microscopy (Olympus, XSZ-D2).
6
Detecting Polysulfides in Cardiomyocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Using the polysulfide-specific fluorescent probe SSP4 (Dojindo, Kumamoto, Kyushu, Japan), the neonatal rat cardiomyocytes were detected for polysulfides, SSP4 (50 μmol/L) probe in serum-free DMEM medium at 37 °C for 30 min. These were washed 3 times with PBS. Fluorescence microscopy detects fluorescence levels. Fluorescence microscopy (Olympus, XSZ-D2) was used to detect the fluorescence responses of polysulfide in the samples.
7
Intracellular Polysulfide Detection in C2C12 Myoblasts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The differentiated C2C12 myoblasts were inoculated in the 24‐well plate to detect the intracellular polysulphide level by fluorescent probe, SSP4, with slight modifications.13 Briefly, cells were loaded with SSP4 (50 μmol/L) in a serum‐free DMEM medium containing 0.003% Cremophor EL for 15 minutes at 37°C in the dark. After being washed, SSP4 was detected using the fluorescence microscope (Olympus, Shenzhen, China, XSZ‐D2).
8
BODIPY-Lipid Uptake in H9c2 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
H9c2 cells were incubated with 2 µmol/L BODIPY 558/568 C12 (Life technologies) and treated with different reagents after 48 hours. Cells were washed three times with DMEM. BODIPY 558/568 C12 fluorescence was determined using fluorescence microscope (Olympus, XSZ‐D2).
9
H2S Detection in Cardiac Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The fluorescence response of H2S in cardiac tissue and neonatal rat cardiomyocytes (NRCMs) was detected using the H2S-specific detection probe C-7Az (7-azido-4-methylcoumarin, Sigma-Aldrich). Frozen sections of db/db cardiac tissues or NRCMs were incubated with H2S-specific detection probe C-7Az (50 μmol/L) in phosphate-buffered saline (PBS) for 30 min at 37 °C. These were washed three times with PBS. Fluorescence microscopy (Olympus, XSZ-D2, Tokyo, Japan) was used to detect the fluorescence responses of H2S in the samples.
10
Visualizing Autophagic Vacuoles in Cardiac Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The frozen sections of mouse hearts and H9C2 cells were incubated with 50 μM MDC in PBS at 37 °C for 30 min. Autophagic vacuoles were analysed using confocal laser scanning and fluorescence microscopy (Olympus, XSZ-D2, Tokyo, Japan).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!