Mouse tissue sections were prepared and stained with TO-PRO-3 Iodide (Invitrogen) to visualize nuclei and avoid interference with the fluorescence spectrum of transferred cell trackers. Single-fluorochrome controls were utilized to ensure no cross-bleeding was present in between fluorescent channels. Images were acquired at 12-bit depth, 1024×1024 pixel size, at 400× and 630× magnifications utilizing either the SP5 Tandem Scanner Spectral 2-photon confocal microscope or the SP8 3D 3-color STED laser scanning confocal microscope with time gating (Leica). Each region of interest (ROI) was 144.74 μm/1024 pixels wide, corresponding to an average absolute resolution size of 0.28 μm, based on Nyquist sampling. Regions of interest, containing all three transferred cell populations, were selected for acquisition. Raw images were stored in manufacturer-specified .lif format. Lif files were converted to multi-channel .tif images and used as input for DCNN analysis.
Sp8 3d 3 color sted laser scanning confocal microscope
Manufactured by Leica
The Leica SP8 3D 3-color STED laser scanning confocal microscope is a high-performance imaging system designed for advanced microscopy applications. It features a combination of confocal and STED (Stimulated Emission Depletion) technologies, enabling high-resolution, three-color imaging of samples.
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Lab products found in correlation
2 protocols using sp8 3d 3 color sted laser scanning confocal microscope
1
Multicolor Confocal Imaging of Transferred Cells
2
Multicolor Confocal Imaging of Transferred Cells
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Mouse tissue sections were prepared and stained with TO-PRO-3 Iodide (Invitrogen) to visualize nuclei and avoid interference with the fluorescence spectrum of transferred cell trackers. Single-fluorochrome controls were utilized to ensure no cross-bleeding was present in between fluorescent channels. Images were acquired at 12-bit depth, 1024×1024 pixel size, at 400× and 630× magnifications utilizing either the SP5 Tandem Scanner Spectral 2-photon confocal microscope or the SP8 3D 3-color STED laser scanning confocal microscope with time gating (Leica). Each region of interest (ROI) was 144.74 μm/1024 pixels wide, corresponding to an average absolute resolution size of 0.28 μm, based on Nyquist sampling. Regions of interest, containing all three transferred cell populations, were selected for acquisition. Raw images were stored in manufacturer-specified .lif format. Lif files were converted to multi-channel .tif images and used as input for DCNN analysis.
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