Sectioning of the paraffin-embedded kidney tissue after dewaxing and rehydrating as shown above were performed with a TUNEL assay following the manufacturer’s protocols (Roche Diagnostics, Mannheim, Germany) as described [41 (link)]. The images were visualized under a Ti2-A fluorescence microscope (NIKON, Tokyo, Japan).
Ti2 a fluorescence microscope
Manufactured by Nikon
Sourced in Japan
The Ti2-A fluorescence microscope is a laboratory equipment designed for high-performance fluorescence imaging. It features advanced optics and illumination systems to enable detailed observation and analysis of fluorescently labeled samples.
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6 protocols using ti2 a fluorescence microscope
1
TUNEL Assay on Kidney Sections
2
Immunofluorescence Analysis of NRF2 in Renal Cells
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Renal tubule epithelial cells were treated with liquiritigenin (50 µM) for 6 h, and then washed with phosphate-buffered saline (PBS, pH = 7.2). Immunofluorescent staining was performed as described [41 (link)]. The images were visualized under a Ti2-A fluorescence microscope (NIKON, Tokyo, Japan). The antibody used for immunofluorescence staining was anti-NRF2 antibody (1:100, Proteintech, Chicago, IL, USA). Quantitative data from at least 30 cells were counted per group from representative triplicate experiments.
3
H&E Staining of Mouse Kidney Tissue
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Four-micrometer mice kidney tissue sections were stained with an H&E reagent after dewaxing and rehydrating in the program as described [41 (link)]. Pathological section images were obtained using a Ti2-A fluorescence microscope (NIKON, Tokyo, Japan).
4
Mitochondrial Membrane Potential Measurement in Renal Cells
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Probe JC-1 staining was used to measure the mitochondrial membrane potential in renal tubule epithelial cells. Post-treatment, medium/JC-1 working solution (1:1) was added to the cell slides in the plate and incubated for 20 min. The staining solution was removed, and then the cells were gently washed twice with JC-1 staining buffer. The pictures were captured by a Ti2-A fluorescence microscope (NIKON, Tokyo, Japan) [43 (link)].
5
Immunohistochemical Detection of SIRT3
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Immunohistochemical staining was performed as described [43 (link)]. Paraffin-embedded kidney tissue sections were stained with anti-SIRT3 antibody (1:100, Abcam, Cambridge, UK) at 4 °C overnight. Graphs were obtained by using a Ti2-A fluorescence microscope (NIKON, Tokyo, Japan).
6
NRF2 Immunofluorescence Assay in Caco-2 and A549 Cells
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The Caco-2 and A549 cells were treated with liquiritigenin (50 μM) for 6 h, washed with phosphate-buffered saline (PBS, pH = 7.2), and fixed. Then, these cells were stained with anti-NRF2 antibody (1:100; Proteintech, Chicago, IL, USA) after blocking with 1% bovine serum albumin (Sigma-Aldrich, St. Louis, MO, USA) in PBS for 30 min at room temperature, followed by washing the cells four times with PBS. Finally, fluorescence-label secondary antibody was added, and the cells were embedded into gelatine solution. Immunofluorescence staining was performed using a Ti2-A fluorescence microscope (NIKON, Tokyo, Japan).
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