SNU-C5RR cells (1×105 cells/mL) were cultured in chamber slides and maintained for 24 h. 38 μM Shikonin was added, and cells were incubated for 24 h. The slides were then prepared following the manufacturer’s instructions for the DeadEnd Colorimetric TUNEL system (Promega Corporation, Madison, WI, USA). Faramount aqueous mounting medium (Agilent Technologies, Inc., Santa Clara, CA, USA) was used to mount stained cells on microscope slides. The microscopic images were captured via confocal fluorescent microscopy using an appropriate Laser Scanning Microscope 5 PASCAL program (Carl Zeiss AG, Oberkochen, Germany).
Laser scanning microscope 5 pascal program
Manufactured by Zeiss
Sourced in Germany
The Laser Scanning Microscope 5 PASCAL program is a precision imaging tool that uses a focused laser beam to scan and capture high-resolution images of microscopic samples. It provides accurate and detailed visualization of the structure and properties of the observed specimens.
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2 protocols using laser scanning microscope 5 pascal program
1
Shikonin-Induced Cell Apoptosis Assay
2
Evaluation of Intracellular ROS Generation
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Cells were seeded in 6 well plates at a density of 3 × 105 cells/ml. After 24 h of incubation at 37°C, the cells were treated for 24 h with 140 μM 5-FU. Cells were treated with 20 μM dihydrorhodamine (DHR) 123, and the DHR123 fluorescence was detected using a flow cytometer (Becton Dickinson, Mountain View, CA, USA) and analyzed using Cell Quest software. For image analysis of the generation of intracellular ROS, cells were seeded on a coverslip-loaded 6-well plate at a density of 2 × 105 cells/ml. After addition of 20 μM DHR123 to each well and incubation for an additional 30 min at 37°C, plates were washed with PBS, and stained cells were mounted onto a microscope slide in mounting medium (DAKO, Carpinteria, CA, USA). Microscopic images were analyzed using the Laser Scanning Microscope 5 PASCAL program (Carl Zeiss, Jena, Germany) on a confocal microscope.
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