Oocytes after IVM for 42 h were selected randomly from the control group and melatonin group, and were incubated in pre-warmed maturation medium at 39 °C and 5% CO2 for 20 min. A Mito Tracker Green kit (Beyotime Institute of Biotechnology, Haimen, China) was used to label distribution of mitochondria of oocytes at 37 °C for 30 min. The labeled oocytes were checked using a fluorescence microscope (Olympus, BX60, Tokyo, Japan), and all oocytes were photographed using a digital camera (Nikon 990, Tokyo, Japan). There were two main distribution features of mitochondrial distribution patterns in oocyte: one was that labeled mitochondria were distributed evenly throughout ooplasm (homogeneous, Figure 1 a), which indicated that mitochondrial distribution was better in oocytes. Others were that the labeled mitochondria were distributed unevenly within ooplasm (heterogeneous; Figure 1 b,c) as described previously [19 (link)]. The abnormal distribution of mitochondria has negative effects on ATP distribution and embryogenesis [20 (link)]. The value of mitochondrial distribution was analyzed in a blind manner, and was determined as the number of oocytes with homogeneous mitochondria/total number of the oocytes × 100.
Mito tracker green kit
Manufactured by Beyotime
Sourced in China
The Mito-Tracker Green kit is a fluorescent probe designed for the detection and visualization of mitochondria in live cells. The kit provides a simple and efficient method for labeling mitochondria, allowing for the analysis of mitochondrial function and dynamics.
Automatically generated - may contain errors
Lab products found in correlation
Ros assay kit, by Beyotime (3 mentions)
Reverse transcription kit, by Accurate Biology (2 mentions)
Trizol, by Thermo Fisher Scientific (2 mentions)
Jc 1 kit, by Beyotime (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Apoptosis detection kit, by BD (2 mentions)
Chamq sybr qpcr master mix kit, by Vazyme (1 mentions)
Nrf2 inhibitor ml385, by MedChemExpress (1 mentions)
Fitc dextran, by Merck Group (1 mentions)
Genomic dna extraction kit, by Omega Bio-Tek (1 mentions)
Cell counting kit 8 cck 8, by Beyotime (1 mentions)
Ros detection kit, by Beyotime (1 mentions)
Atp assay kit, by Beyotime (1 mentions)
Mmp assay kit, by Beyotime (1 mentions)
Dmem high glucose medium, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle medium, by Thermo Fisher Scientific (1 mentions)
D8417, by Merck Group (1 mentions)
Image pro, by Media Cybernetics (1 mentions)
Fluoview fv1200, by Olympus (1 mentions)
Accuri c6, by BD (1 mentions)
7 protocols using mito tracker green kit
1
Evaluating Mitochondrial Distribution in IVM Oocytes
2
Nrf2-mediated Antioxidant Response Assay
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Sodium selenite, fluorescein isothiocyanate-dextran (FITC-dextran) with average molecular weight 4,000 and H2O2 were purchased from Sigma-Aldrich Co.. Nrf2 inhibitor (ML385) was purchased from MedChemExpress (Shanghai, People's Republic of China). All reagents for cell culture were purchased from Life Technologies (Thermo Fisher Scientific, Waltham, MA, USA). Mito-Tracker Green Kit, ATP Assay Kit, MMP Assay Kit, ROS detection Kit, and Cell Counting Kit-8 (CCK-8) were purchased from Beyotime Biotechnology (Shanghai, People's Republic of China). RIPA lysis buffer and bicinchoninic acid (BCA) protein assay kit were purchased from Solarbio Life Sciences Biotech Co. (Beijing, People's Republic of China). Nrf2, NQO-1, and HO-1 primary antibodies and corresponding secondary antibody were purchased from Abcam Biotechnology (Cambridge, MA, USA). Genomic DNA extraction Kit was purchased from OMEGA Bio-Tek (Norcross, GA, USA). ChamQTM SYBR® qPCR Master Mix Kit was purchased from Vazyme (NanJing, People's Republic of China).
3
Mitochondrial Function and Oxidative Stress Assay
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DMEM high-glucose medium and FBS were purchased from Gibco. The Mito-Tracker Green kit and JC-1 kit were purchased from Beyotime. The total RNA extraction reagent Trizol was purchased from Life Technologies. The rabbit anti-monoclonal antibody and HRP-labeled mouse anti-rabbit IgG were purchased from CST. The ROS assay kit was purchased from Beyotime, the apoptosis detection kit was purchased from BD, the reverse transcription kit and qPCR kit were purchased from Accurate Biology, and primers were synthesized by Accurate Biology according to the designs detailed in Table 1 .
4
Mitochondrial Dynamics in C2C12 Myoblasts
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Mouse C2C12 myoblast cell lines were obtained from the Shanghai Institute of Cell Biology, Chinese Academy of Sciences. Dulbecco’s Modified Eagle Medium, a high-glucose medium, and fetal bovine serum (FBS) were purchased from Gibco. The Mito-TrackerGreen kit and JC-1 kit were purchased from Beyotime. Total ribonucleic acid (RNA) extraction reagent TRIzol was purchased from Life Technologies. Antibodies against Drp-1 and myostatin were obtained from Cell Signaling Technology. The ROS assay kit was obtained from Beyotime, and the apoptosis detection kit was purchased from Becton, Dickinson, and Company. The reverse transcription kit and quantitative polymerase chain reaction (qPCR) kit were purchased from Accurate Biology.
5
Mitochondrial Content Quantification
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Mitochondrial content was assessed using the Mito-Tracker Green kit (C1048; Beyotime Institute of Biotechnology). After treatments were imposed as indicated in the Cell Culture and Treatments section, MAC-T cells were incubated with MitoTracker Green (200 nM) in serum-free medium at 37°C for 30 min, followed by fixing in 10% formaldehyde neutral buffer solution at room temperature for 30 min. Cells were then stained with DAPI (10 μg/mL; D8417, Sigma-Aldrich) at room temperature for 10 min. Cells were imaged using laser confocal microscopy (Fluoview FV1200, Olympus). Fluorescence images were analyzed using the Image-Pro image analysis software (Media Cybernetics) as described in the Mitochondrial Membrane Potential section. Mitochondrial content was quantified by mean density (i.e., mean staining intensity = integrated optical density sum/area sum).
6
Mitochondrial Dynamics in Cancer Cells
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A549 cells were treated with mPEG-SS-PGG-SS-IR780/PTXL MCs and mPEG-PGG-IR780/PTXL MCs with or without NIR. After 4 h, the cells were collected to evaluate the mitochondrial localization, mitochondrial membrane potential (MMP), ROS, and ATP contents by using a Mito-tracker green kit (Biyuntian, Hangzhou, China), an MMP assay kit with JC-1 (Elabscience, Wuhan, China), an ROS assay kit (Biyuntian, Hangzhou, China), and an ATP assay kit (Solaibao, Beijing, China), respectively.
7
Mitochondrial Quantification in Pancreatic Cells
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The mitochondrial quantification experiment was performed using a Mito-Tracker green kit (Biyuntian, Nanjing, China), following a standard procedure. The pancreatic single cells were first suspended with dye buffer at 106 cells/mL and incubated at 37 °C for about 30 min. Then, Mito-Tracker Green Dyeing buffer was removed by centrifugation at 500× g for 5 min. The cells were washed twice with PBS, and the cells were resuspended in fresh culture medium at 37 °C. Then the cell fluorescence was detected by Accuri C6 (BD). The fluorescence intensity represents the relative mitochondrial number.
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