Treated cell smears were naturally dried and fixed at room temperature for 30 min using 4% paraformaldehyde. After washing with PBS three times, samples were immersed in the sealing solution (3% H2O2, methyl) and blocked at room temperature for 10min. Then, samples were immersed in cell membrane permeabilization solution (0.1% Triton X‐100) for 2 min at 37℃. Next, samples were stained with TUNEL Apoptosis Detection Kit (Roche, Germany) and counterstained with DAPI. Finally, a fluorescence microscope system (Carl Zeiss, Germany) was utilized to capture the images.
Fluorescence microscope system
Manufactured by Zeiss
Sourced in Germany
The Fluorescence Microscope System from Zeiss is a specialized optical instrument designed to capture and analyze the fluorescence properties of samples. It utilizes a focused beam of light to excite fluorescent molecules within the specimen, which then emit light at a different wavelength. This allows researchers to visualize and study the distribution, interactions, and dynamics of specific labeled components within the sample.
Automatically generated - may contain errors
Lab products found in correlation
Tunel apoptosis detection kit, by Roche (2 mentions)
Tunel kit, by Roche (2 mentions)
Lucia software, by Nikon (1 mentions)
Biotin 16 dutp, by Roche (1 mentions)
Anti lamp1, by BD (1 mentions)
Anti synaptophysin, by Merck Group (1 mentions)
Anti calbindin d 28k, by Merck Group (1 mentions)
Anti glur1, by Merck Group (1 mentions)
Anti cathepsin b, by Merck Group (1 mentions)
C2 point scanning confocal microscope, by Nikon (1 mentions)
6e10 antibody, by Fortrea (1 mentions)
Nis elements ar software, by Nikon (1 mentions)
8 protocols using fluorescence microscope system
1
Apoptosis Detection in Cell Smears
2
TUNEL Assay for RCC Apoptosis
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RCC cell apoptosis was examined using terminal deoxynucleotidyl transferase-mediated dUTP nick-end-labeling assay (TUNEL) kit (Roche) according to the manufacturer's instruction. After TUNEL staining, images were captured by a Fluorescence Microscope System (Carl Zeiss).
3
CCK-8 Assay for Proliferation Analysis
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For the CCK-8 assay, CRC cells were suspended in 96-well plates at 1 × 106 cells/well, 20 μL of the thymidine analog5-ethynyl-29-deoxyuridine (EdU) solution (Becton-Dickinson, USA) was co-incubated with cells, the results were recorded by using the fluorescence microscope system (Carl Zeiss, Germany). The morphological change of CRC cells treated with anlotinib was observed with NIS microscope imaging software (Nikon, Japan). EdU-positive staining cells be counted in five diverse areas of each well, and we implemented three independent experiments in triplicate.
4
Photothermal Therapy of Tumor-Bearing Mice
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Balb/c nude mice (5 weeks old) were obtained from the Laboratory Animal Center of Nanjing Medical University (Nanjing, China). The animal procedures were in accordance with the guidelines of the Institutional Animal Care and Use Committee of Nanjing University. EMT-6 cells (1×107 cells/mL, 100 μL) were inoculated subcutaneously into the back of a nude mouse. Once the volume approached 100 mm3, the mouse was anaesthetized with pentobarbital sodium (i.p., 60 mg/kg) and intravenously injected with 100 μL of the NPs (10 mg/mL). Twelve hours later, the mouse was treated with an 808 nm laser in situ (0.8 W/cm2) for 10 mins. During the treatment, temperature at the tumor was recorded with a thermal imager and the luminescence of NPs was detected with a cooled visible light CCD (Sony ICX694, Japan) through a 500–550 nm bandpass filter. At 6 hrs after the thermal therapy, the treated mouse was imaged with the CCD camera, and then the tumor tissues and organs of the mouse were excised, fixed in 10% paraformaldehyde solution and subsequently processed routinely into paraffin. The mouse for control was treated as above but no injection and irradiation. All the sliced tissues were stained with H&E and examined by a Zeiss fluorescence microscope system.
5
TUNEL Apoptosis Assay for Kidney Cells
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Apoptosis of the PTEC cells or frozen kidney sections was examined using a TUNEL kit (Roche, Germany) according to the manufacturer’s instructions. Briefly, the samples were fixed with 4% PFA, permeabilized with 0.5% Triton X-100 in PBS, and blocked with 5% bovine serum album (BSA). Then, the cells or tissue sections (5 μm) were stained with the TUNEL Apoptosis Detection Kit (Roche, Germany) and counterstained with DAPI. After staining, the images were captured by a fluorescence microscope system (Carl Zeiss AG, Germany).
6
Histological and ROS Analysis of Murine Kidney
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For the morphometric analyses, transverse tissue sections (5 μm) of formalin-fixed and paraffin-embedded murine kidney tissues were stained with a standard hematoxylin and eosin (H&E) procedure. LUCIA software (Nikon) was used to capture the images.
Dihydroethidium (DHE, Cat No. S0063, Beyotime, China) was used to examine reactive oxygen species (ROS). Briefly, transverse tissue sections (5 μm) were subjected to DHE staining according to the manufacturer’s instructions, and the images were captured by a fluorescence microscope system (Carl Zeiss AG, Germany).
Dihydroethidium (DHE, Cat No. S0063, Beyotime, China) was used to examine reactive oxygen species (ROS). Briefly, transverse tissue sections (5 μm) were subjected to DHE staining according to the manufacturer’s instructions, and the images were captured by a fluorescence microscope system (Carl Zeiss AG, Germany).
7
Chromosome Characterization of Ae. biuncialis Introgression Lines
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Root tips of germinating seeds from the Mv9kr1ph1b_K × Ae. biuncialis amphiploids containing chromatin introgressed from Ae. biuncialis accessions MvGB380, MvGB1714, MvGB1733, MvGB1987, and MvGB1723 were used for chromosome preparation as described by Lukaszewski et al. (2004) (link). Genomic in situ hybridization (GISH) experiment was done as described by Molnár et al. (2009) (link). Briefly, total genomic DNAs of Ae. umbellulata (UU) and Ae. comosa (MM), the diploid progenitors of Ae. biuncialis, were labeled with biotin (biotin-16-dUTP; Roche) and digoxigenin (digoxigenin-11-dUTP; Roche) by random priming and used as U- and M-genome probes, respectively. Unlabeled wheat genomic DNA was used as blocking DNA at a ratio of 30:1. Digoxigenin and biotin signals were detected using anti-digoxigenin-rhodamine Fab fragments and Alexa Fluor488, respectively. The slides were evaluated using the Zeiss fluorescence microscope system as described for the meiotic chromosome pairing analysis.
8
Immunohistochemical Analysis of Alzheimer's Markers
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Fixed tissue was cryoprotected and serially sectioned at a thickness of 10–20 μm. The tissue sections were immunolabeled using 6E10 antibody (Covance), anti-calbindin D28k (Sigma-Aldrich), anti-GluR1 (from Millipore or developed as described [88 (link)]), anti-LAMP1 (BD Pharmingen; San Jose, California, USA), anti-cathepsin B (Millipore), anti-synaptophysin (Millipore), and antibodies that selectively label Aβ38 and Aβ42 (Covance). Immunofluorescence analyses used appropriate Invitrogen secondary antibodies (Thermo Fisher Scientific; Waltham, Massachusetts, USA), and images were captured with a Zeiss fluorescence microscope system (Carl Zeiss, Inc.; Thornwood, New York, USA) and with a Nikon C2 point-scanning confocal microscope with NIS-Elements AR software (Nikon Instruments; Melville, New York, USA). Other images were produced via avidin–biotin–peroxidase protocols (Vector Laboratories; Burlingame, California, USA) that used 3,3′-diaminobenzidine as the chromogen. In each case, treatment groups were immunostained together and analyzed under the same instrument settings. Equally spaced coronal sections along the rostral–caudal axis of the hippocampus were used to determine the average immunoreactivity intensity and area of staining across four different view-fields.
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