Transcript Celigo Key Features

Celigo is a multi-channel imaging cell cytometer designed for scientists performing bench-top experiments using suspension and non trypsinized adherent cells in micro-well plates.

Performing an assay using Celigo can be accomplished in a few easy steps. Additional features highlighted in this video include image autofocus, five-color acquisition, imaging and analysis of every cell in the well, instrument speed, exporting of data and images, and gating analysis.

First, we insert the plate into the Celigo. In this example, each well contains a varied population of A549 cells with expression of green or red fluorescing proteins.

Select well C4 to set up focus.

Find the focus location by using Celigo’s Image-based Auto-focus function, then apply an offset to obtain “Ideal cell counting focus”.

For the rest of the plate, each well is auto-focused using the same focus settings.

For each well of a 96-well plate, there is only one physical movement and one focus event.

Celigo system consists of scanning optics that capture bright field and fluorescent images.

Uniform illumination and focus are maintained all the way to the well edge, allowing us to count all the cells in each well.

Celigo is able to scan and perform image analysis concurrently.

Depending on the number of fluorescent channels the scanning time ranges from 5-12 minutes per 96-well plate.

Results for each well are displayed immediately in a plate map.

From the plate map click on well D5 to zoom in on the whole well image. We can view the overlay of bright field and GFP, a GFP only, bright field and RFP, or a RFP only images.

Export numerical data for the whole plate into Excel in csv format for data analysis and graphing.

Export high-resolution cell images from each well either as one whole image or as multiple sections.

Image format include tiff, jpg, and bitmap, allowing analysis by other software packages.

For population analysis, the gating function is used for histogram and 2D scatter plots.

In well D6, GFP and RFP mean fluorescence intensity for each cell is displayed in the scatter plot.

Cells belonging to a particular quadrant of the plot are circled in corresponding colors.

Same gating set up is then applied to other wells during analysis.

In summary, the Celigo instrument possesses an easy work flow, image auto focus, four color acquisition, imaging and analysis of every cell to the well edge, fast image scanning and analysis, ability to export of data and images, and perform population histogram and 2D gating analysis.

Additional Celigo videos can be found at www.nexcelom.com/Celigo

Free on-site technical seminars or demonstrations are provided for your lab.