Introduction to FCS Express 5 Image Cytometry
Contents
A Flow Cytometry environment for Image Cytomtery Data
FCS Express 5 Image Cytometry is a new version of FCS Express that contains exciting and unique tools for working with Image Cytometry data.
This tutorial is designed to provide an overview of many powerful new tools that allow you to work with image data in much the same way as flow cytometry data. Now you can combine population based analysis and single cell interrogation of your image cytometry results in one software package!
This three section tutorial is designed to take you through prepared examples demonstrating the use of FCS Express 5 Image Cytometry.
This tutorial closely follows the "Introduction to FCS Express 5 Image Cytometry" video on the recorded webinars page of the De Novo Software website. It is recommended you watch the video PRIOR to starting the tutorial. You will better understand the overview and biology behind the data by doing so.
In the course of the tutorial you will load and review image datasets, establish and modify gating strategies and view individual cells in cell galleries, use the High Content Add-on to optimize and view a 96-well plate nuclear translocation assay. The High Content Add on enables review of multi-parametric results and images from an entire plate.
A brief example of Batch analysis and FCS Express reporting tools will also be provided. This tutorial will not expand on the details of many common features and tools in FCS Express 5 for which there are existing detailed tutorials. When relevant, tutorials designed to drill down and teach those commonly used functions will be referenced.
In the course of this multi-part tutorial, we will specifically:
| • | Demonstrate loading image cytometry data and working with images |
| • | Review gated populations in data grid with cell image galleries |
| • | Review analysis and reporting tools for image cytometry |
| • | Optimize the assay settings for a nuclear translocation assay |
| • | Use the "High Content Analysis" module to work with an image-based assay in a 96-well plate |
| • | Review batch analysis and reporting option for image cytometry data |
Due to the large size of image data, we have only worked with a several example images for each part of the tutorial. The images are embedded in the example layouts. We do not provide the image data from an entire 96-well plates in our Tutorial Sample Data Archive for examples using the High Content Add on. If you would like to move beyond this introductory tutorial and work with data from the whole study please use our step by step instructions in the CellProfiler Export Tutorial.