The morphological changes of apoptotic cells were observed by Hoechst 33258 staining. The cells were placed in 6-well plates at 8 × 104 cells/mL, exposed to the drug(s) for 48 h, and washed twice with ice-cold PBS. Cells were fixed with fixative consisting of methanol and glacial acetic acid (3:1, v/v) for 15 min, followed by Hoechst 33258 staining (10 mg/L) (Beijing Solarbio Science & Technology Co., Ltd., Beijing, China) for 10 min at room temperature and away from the light, as described previously (Soriano et al., 2013 (link)). Apoptosis morphology was captured using a fluorescence microscope (MIC00266; Zeiss, Germany) and analyzed by Image-Pro Plus Premier (version 9.1.4; Media Cybernetics Inc., Rockville, MD, United States).
Image pro plus premier
Manufactured by Media Cybernetics
Sourced in Germany, China
Image-Pro Plus Premier is a comprehensive software package designed for advanced image analysis and processing. It offers a wide range of tools and features for researchers, scientists, and professionals working with digital images.
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Lab products found in correlation
2 protocols using image pro plus premier
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Hoechst 33258 Staining of Apoptotic Cells
2
Apoptosis Assay of T24 Cells
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The T24 cells were seeded in 6-well culture dishes (2×105 cells/well). Following treatment with ISL at 0, 30, 50 and 70 µg/ml at 37°C for 24 h, the cells were fixed with Carnoy's fixative consisting of methanol and glacial acetic acid (3:1, v/v), washed twice with PBS, stained with Hoechst 33258 (5 mg/ml) for 5 min in the dark, as described previously (28 (link)), and washed extensively three times with PBS. Nuclear staining was examined using a fluorescence microscope (Nikon LH-M100CB; Jirui Co., Ltd., Suzhou, China) and images were captured using Image-Pro Plus Premier (version 9.1.4; Media Cybernetics, Inc., Rockville, MD, USA). Cells that exhibited decreased nuclear size, chromatin condensation, intense fluorescence and nuclear fragmentation were considered apoptotic.
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