Celigo FAQs
How many channels are available?
- 1 brightfield channel
- 3 fluorescent channels
- Blue: 385 nm LED (377/50 excitation, 447/60 emission filters)
- Green: 464 nm LED (483/32 excitation, 536/40 emission filters)
- Red: 528 nm LED (531/40 excitation, 629/56 emission filters)
- Fluorescent channels can be imaged individually or in combination
- Fluorescent channels can be combined with brightfield
What plate types are recommended for use on Celigo?
- Efficient imaging on Celigo requires plates which have:
- high optical quality for even illumination, imaging, and segmentation
- SBS-standard foot print with proper well definition to ensure full well processing
- We continue to expand our plate offerings. New plate formats and types are released continuously.
| Plate | Vendor | Catalog # |
| 1536-well | Corning | 3832 |
| 384-well | C-lect™ | 400101 |
| C-lect™ | 400102 | |
| Corning | 3712 | |
| 96-well | C-lect™ | 400103 |
| C-lect™ | 400104 | |
| Corning | 3603 | |
| 24-well | Corning | 3524 |
| Corning | 3338 | |
| PerkinElmer | 145060X | |
| 12-well | C-lect™ | 400402 |
| Corning | 3512 | |
| 6-well | C-lect™ | 400401 |
| Corning | 3516 | |
| Corning | 3471 |
What is the format of data files generated by Celigo?
- Numerical data files are formatted as comma separated value (csv) files
- Images may be generated in bmp, jpeg and tiff formats
- Data files may be exported in FCS or Multiple FCS for use in flow cytometry software such as FlowJo and FCS Express
What is the file size for one plate single channel compared to multiple channels?
- 1 channel datasets are approximately 1.5GB per plate. Accordingly, a 2 channel experiment would require ~3GB. This is generally independent of plate type as one 96 well contains 16 fields/well for a total of 1,536 images and one 384 well contains 4 fields/well for the same total of 1,536 images.
Can I analyze my data with any other scientific/statistical software?
- Numerical data is exported in a comma separated value (csv) format for analysis in programs like Excel, GraphPad Prizm and OpenOffice.org Calc.
- Image files may be analyzed with any software that accepts bmp, jpeg or tiff files (MetaMorph, Image-Pro Plus, CellProfiler).
- Data files may be exported in FCS or Multiple FCS for use in flow cytometry software such as FlowJo and FCS Express.
Can I analyze my data on another computer other than the Celigo computer?
- Cyntellect offers a Celigo Satellite Workstation that can be used for offline processing of Celigo data.
Can I connect the Celigo database to my server with network database functionality?
- No, the Celigo does not have network database functionality at this time.
What is the difference between manual and auto-registration of the hardware autofocus
- Manual registration- This method of registering the focus allows the user to quickly define the best focus for a given scan manually. The registration is not saved and therefore cannot be loaded for subsequent scans.
- Auto registration – The software automatically determines the focus registration using an image-based algorithm (which may be adjusted manually using an offset). Using this method the focus registration is stored and may be used for batch analysis.
What type of robots can be connected to the Celigo?
- Cyntellect offers an automation application programming interface (API) for the Celigo that enables automated plate loading, scanning and processing using a robotic arm such as the Plate-Crane or Hamilton Star.
How do I accurately segment cells in brightfield?
- Accurate segmentation is dependent on exposure and focus settings that give the best contrast.
- In general, cells with larger amounts of cytoplasm, such as HeLa’s segment better using a "bright focus" and those that are flatter with less cytoplasm (epithelial cells) may require a "dark focus". Please see the Celigo Users Guide for details on each focus type.
What is the difference in the 3 segmentation algorithms used for Expression Analysis?
- Independent: Each channel is segmented and processed separately. Subpopulations of cells may be present in one channel and absent in another, therefore, data is only acquired for cells in channels where they emit a positive signal. Negative cells are not included in the analysis.
- Mask: Data is acquired in all channels for cells detected and segmented in the “Mask” channel only. If the entire population possesses the “Mask” marker, all cells will be included in the analysis.
- Merge: Each channel is segmented separately and processed separately. The objects overlays are then merged to create a single mask. Cells segmented in different channels are processed with this merged single mask to include all cells in the analysis even if they have signal in only one channel.
Can I analyze sub-cellular structures?
- The high resolution of Celigo allows for accurate analysis of the majority cell based assays.
- At a 1um/pixel resolution, sub-cellular structures, including nuclei and cytoplasm, can be readily analyzed. However, some small organelles may not be resolved accurately on Celigo.
Can I analyze and count colonies?
- Whole-well imaging on Celigo allows colonies to be efficiently imaged using a wide range of plates.
- Currently, cells growing in colonies can be segmented and analyzed to evaluate viability, expression levels, etc (see marker expression).
- At this time, with the exception of sphere cultures, whole colonies can not segmented and counted on Celigo. However this is currently in development.
Can I analyze small organisms?
- The large imaging area of Celigo is ideal for imaging small organisms; however, the analytical software is not currently available.
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