Operator Workflow - Counting Cells

Form

The input form ZEISS Cells is provided. The form is displayed in the report.

The default form is displayed.

To change the form, in the Form Selection tool, select the form, see Form Selection Tool.

Fill in relevant data according to your sample.

2D Multi-Channel Acquisition

Acquire images.

To acquire multi-channel images, open Smart Setup and add dyes and contrast methods according to your sample. The first channel should be the fluorescently-labeled nuclei. Smart Setup provides proposals setting the light source, camera and filter setting, see Configuring Multi-Channel Acquisition with Smart Setup.

Adjust the acquisition parameters, e.g. measure the exposure time, and then take a snap.

Image Segmentation

Generate one mask for all nuclei per cell.

Generate masks for nuclei, which are then counted, see Automatic Segmentation.

To separate neighboring nuclei use a separation tool, e.g. Watersheds.

If a large part of the image is selected initially, use background subtraction.

Region Filter

Results are refined based on area and circularity of the objects.

To refine the selection to limit the analysis to one object or nucleus per cell, use region filters for area and circularity, see Region Filter.

Measurement Data

Check the results based on the previously conducted image analysis. The results are displayed in a table.

To select an object, click a value in the table, and to check the size of a certain object, click the object in the Center Screen Area.

For more information, see Measurement Data Tool.

Report

Creates a report document, see Reports. You can print the report.

The report contains the following information:

• Form
• Counts and counts/mm² for each image and their mean
• Display of all images with the detected nuclei

For your documentation, you create a report.