The protocol for adherent mammalian cell lines was optimized for Stellaris FISH probes. Hybridization was performed overnight and no anti-fade was used for imaging. The sequence of the CAL Fluor Red 590 tagged probes targeting the CYP19A1 mRNA can be provided upon request. Samples were imaged using a Nikon Ti-E scanning laser confocal inverted microscope (A1) with 60x oil objective in tandem with Nikon NIS-Elements imaging software. Excitation was by 561.5 nm diode-pumped solid state. Detection was via 595-50 nm filter. Optical sections were captured at 0.300 μm intervals and a resolution of 256 by 256 pixels and zoom factor of 6.8, resulting in a voxel size of 0.0047 μm3 (0.1243 μm by 0.1243 μm by 0.3 μm). Four times averaging was used to reduce photon and camera noise. An automated spot count algorithm determined the number of mRNA 32 (link). For the analysis we included 30 positive control cells to better define an mRNA spot.
Ti e scanning laser confocal inverted microscope a1
Manufactured by Nikon
The Ti-E scanning laser confocal inverted microscope (A1) is a highly advanced imaging system designed for precise and detailed microscopic analysis. It utilizes a laser-scanning technique to capture high-resolution, three-dimensional images of samples. The core function of this equipment is to provide researchers and scientists with a powerful tool for conducting detailed, non-invasive examinations of various specimens.
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Lab products found in correlation
3 protocols using ti e scanning laser confocal inverted microscope a1
1
Optimized Adherent Cell Imaging Protocol for Stellaris FISH
2
Confocal Microscopy Imaging Optimization
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Samples were imaged using a Nikon Ti-E scanning-laser confocal inverted microscope (A1) with a 60× oil objective in combination with Nikon NIS-Elements imaging software. Excitation was by 561.5-nm diode-pumped solid-state and 402.1-nm diode lasers. Detection was via 595-50 nm and 450-50 nm band-pass filters. Optical sections were captured at 0.300-μm intervals and a resolution of 256 by 256 pixels and a zoom factor of 6.8, resulting in a voxel size of 0.0047 μm3 (0.1243 μm × 0.1243 μm × 0.3 μm). Averaging was used four times to reduce photon and camera noise.
3
Optimized Adherent Cell Imaging Protocol for Stellaris FISH
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The protocol for adherent mammalian cell lines was optimized for Stellaris FISH probes. Hybridization was performed overnight and no anti-fade was used for imaging. The sequence of the CAL Fluor Red 590 tagged probes targeting the CYP19A1 mRNA can be provided upon request. Samples were imaged using a Nikon Ti-E scanning laser confocal inverted microscope (A1) with 60x oil objective in tandem with Nikon NIS-Elements imaging software. Excitation was by 561.5 nm diode-pumped solid state. Detection was via 595-50 nm filter. Optical sections were captured at 0.300 μm intervals and a resolution of 256 by 256 pixels and zoom factor of 6.8, resulting in a voxel size of 0.0047 μm3 (0.1243 μm by 0.1243 μm by 0.3 μm). Four times averaging was used to reduce photon and camera noise. An automated spot count algorithm determined the number of mRNA 32 (link). For the analysis we included 30 positive control cells to better define an mRNA spot.
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