Apoptosis was determined with a TUNEL kit (Boster, Wuhan, China) according to manufacturer’s protocol. A negative control was employed by replacing TUNEL reaction mixture with marking fluid without DNA terminal transferase. After counterstaining with hematoxylin, specimens were observed for cell apoptosis. With the use of a light microscope (Leica DM LB, Germany), apoptosis cells presented with yellow solid precipitations in the nucleus and normal cells presented with blue solid precipitations. Apoptosis rates in both groups were recorded and analyzed. LEC apoptosis number was counted by calculating apoptosis rates. The apoptosis rates were calculated by randomly choosing 3 fields in each stretched preparations under a microscope and 100 cells were counted in each field to calculate apoptosis rates.
Tunel kit
Manufactured by Boster Bio
Sourced in China
The TUNEL kit is a laboratory tool used for the detection and quantification of apoptosis, or programmed cell death, in biological samples. The kit utilizes the TUNEL (Terminal deoxynucleotidyl transferase dUTP Nick End Labeling) assay, which labels the DNA strand breaks that occur during the apoptotic process. This allows for the visualization and analysis of cells undergoing apoptosis.
Automatically generated - may contain errors
Lab products found in correlation
Imager a1, by Zeiss (1 mentions)
Hoechst 33342, by Merck Group (1 mentions)
Hematoxylin, by Wuhan Servicebio Technology (1 mentions)
Eclipse 80i, by Nikon (1 mentions)
3 3 diaminobenzidine (dab), by Solarbio (1 mentions)
Dm2000 led, by Leica (1 mentions)
Image pro plus 6, by Media Cybernetics (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Fluorescence microscope, by Olympus (1 mentions)
11 protocols using tunel kit
1
Quantifying Cellular Apoptosis via TUNEL
2
TUNEL Kit for Apoptosis Analysis
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A TUNEL kit (Boster, Wuhan, China) was used for DNA fragmentation analyses. All operations were conducted according to the manufacturer’s protocol. For each slide, five fields were randomly chosen from each section under the microscope. The apoptotic index was determined as the number of apoptotic cells per villus.
3
Analyzing Apoptosis in MIN6 Cells
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After different treatments, MIN6 cells were stained with Hoechst 33342 (Sigma-Aldrich) for 10 min at room temperature, washed with PBS, and then observed using a fluorescence microscope (Imager A1, Zeiss). All the cell nuclei were stained with a blue color. Cells with high blue fluorescence and fragmented nuclei were considered to be apoptotic, while weak blue signal indicated live cells. Images were obtained in random fields, and quantification of apoptotic cells was determined for at least 2000 cells in eight fields for each well. Apoptotic cells in primary islets were measured using a Tunel kit (Boster) and performed according to the manufacturer’s instructions.
4
Quantifying Apoptosis in Mouse Hippocampus
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Pretreated of 6-μm thick paraffin-embedded sections were processed as described for the immunohistochemistry procedure. Apoptosis was determined with the TUNEL kit (Boster, China) according to the manufacturer’s protocol. Section positions were selected as described above (Fig. 1 a). Images of TUNEL staining were captured using an Olympus microscope (DP80, Olympus, Tokyo, Japan) with a × 40 objective. Image-Pro Plus 6.0 software was used to count TUNEL-positive cells in each visual field. Three microscopic fields of the CA1 or CA3 region from each section, three sections from each mouse, and three mice from each group were used. All of the mouse sections used for the analysis were viewed in a blinded manner. We averaged the results for each group.
5
Apoptosis Detection via TUNEL Staining
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The fixed samples were embedded in paraffin, and a 4-μm section was mounted on glass slides and then dried for 12 h at 37 °C. The prepared samples were passed through xylene and ingredient ethanol solutions for deparaffinization, and then immunohistochemistry stained using a TUNEL kit (Boster, Wuhan, China), which is a marker of apoptosis, in accordance with the manufacturer’s protocol.
6
Apoptosis Evaluation in Tumor Tissues
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First, 4% methanol-fixed tumor tissues were sliced into 4 μm thick sections. A TUNEL kit (Boster Biological Technology Co., Ltd., Wuhan, Hubei, China) was utilized to evaluate the apoptosis of tumor tissues in accordance with the manufacturer’s protocols.
7
Myocardial Apoptosis Quantification
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The paraffin samples of the myocardium were prepared and tested by TUNEL kit (MK1025, Boster Biological Technology Co., Ltd., Hubei, China). Apoptotic cells were those with brown or brown-yellow particles. A total of five high-power visual fields were randomly taken from each section. ImageJ software was utilized to calculate the number of TUNEL-positive cells in each group. Apoptosis rate = (number of positive cells/total number of counted cells) × 100%.
8
TUNEL Staining for Apoptosis Detection
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TUNEL staining was performed according to the instructions of TUNEL kit (BOSTER, Wuhan, China). Mouse tumor tissue was taken for paraffin section. Sections were routinely dewaxed and then subjected to section digestion. The specimens were added with labeling buffer and labelled at 37°C for 2 h. Afterwards, blocking solution was added to block for 30 min at room temperature. Biotinylated anti-digoxigenin antibody was diluted with SABC (biohao, Wuhan, China) diluent, and after DAB (Solarbio, Beijing, China) staining, hematoxylin (Servicebio, Wuhan, China) was lightly counterstained. Dehydrated, transparent, sealed and observed under microscope (Eclipse 80i, Nikon).
9
Quantification of Skeletal Muscle Apoptosis
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Paraffin-embedded skeletal muscle sections were assessed with a TUNEL kit (MK1025 Boster, Wuhan, China) as directed by the manufacturer. A light microscope (Leica DM2000 LED, Japan) was utilized for imaging, followed by quantitation with Image J (NIH, USA) and ImagePro Plus 6.0 (Media Cybernetics, USA). Skeletal muscle apoptosis was quantitated by the apoptotic index (amount of apoptotic cells/total amounts of cells). Six randomly selected fields at 200× per sample.
10
Apoptosis Analysis of Rat Hippocampal Neurons
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Paraffin-embedded sections of rat hippocampi were obtained. TUNEL assay was performed using a TUNEL kit (Boster Biological Technology, Pleasanton, CA, USA) according to the manufacturer's protocol. The sections were sealed with natural gum. Apoptotic cells were observed under a light microscope (magnification, ×400) and counted in 6 fields of vision. The apoptosis index was expressed as the number of apoptotic cells within 1 mm2, and the apoptosis of rat hippocampal neurons was defined as the number of apoptotic neurons/total number of neurons ×100%.
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