Spheroid = single cell made in 3D
> Analysis available using JuLI Stage Software
Organoid = making real body parts (using matrix gel)
> Only monitoring
Note: Activation dongle must be inserted at all times to activate Spheroid STAT SW.
*If installing in PCs other than the one provided, codec and DirectX must be installed as well.
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Must use Corning U-bottom plate (Lot. 7007) and make sure to select 96 Well Plate (Spheroid/Scratcher) vessel option. |
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Well calibration: 1. Go to Setup tab and locate the edge of one of the well. 2. Place the cover of the 96 well plate. 3. Adjust focus to make clear distinction between the edge and the background for easier calibration. 4. Click Vessel > Calibration. 5. Proceed with calibration. |
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| When focusing spheroid on brightfield channel, set D-phase value to 3 to make the edge clear. | |
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Adjust fluorescence channel with D-phase value of 5. *Adjust with pseudo color to prevent over-exposure. |
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For spheroid time-lapse, 1. Select auto focus. 2. Click default then Add Position. 3. Select all applicable channels. *If only using brightfield, quick mode can be used for faster result. |
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*Prior to real test, pre-test is recommended to make sure all spheroids are in the center of each well.
Pre-test (only to see position of each spheroid): 1. Use brightfield channel and set focus. 2. Select All > Default > Add Position. 3. Run time-lapse. |
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Spheroid STAT Software |
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1. Set parameters. 2. The most two important settings to adjust are -Fill holes level : to fill in any holes found in spheroid -Shrink level : to remove any included debris around the spheroid |
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Other parameters: -Circularity : the closer to 10, the more round the spheroid will be adjusted. -Background level : the higher the value, the brighter the background is. -Smooth level : smoothing the edge of the spheroid to make it more round. |
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Use the default gating option for fluorescence channel.
*Use the background level if default gating option does not provide an optimal result. *If both cases do not work, adjust image in edit tab
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Diameter, area, volume, circularity and parameter of spheroid are available.
Adjust parameter values for any well with off result.
Use image overaly to add scale bar, values, etc. and save the image. |