Kaluza Linked Gates Tutorial with Dr. David Onion
Linked Gate feature is a useful tool in Kaluza Analysis Software that helps ensure consistency when placing gates that you want to be identical on multiple bivariate plots or indeed on histograms.
Boolean gating allows us to create gates based on a logical selection of other individual gates and with the use of operators such as AND, NOT, and OR. You might want to create a simple combination of 2 gates, right up to a complex selection of multiple gates including or excluding populations as required.
Exploring batch analysis with Kaluza Analysis Software. In a nutshell batch analysis allows you to use any protocol file, composite, or Kaluza analysis file as a template to perform analysis on as many files as you like. This would become really useful if you were compiling reports, perhaps you’re writing a paper and you wanted to get all your data together and make sure it is analyzed in exactly the same way.
Flow cytometry is an excellent way to measure DNA content because of its high sensitivity high precision acquisition of cell by cell total fluorescence, and of course DNA content analysis was the original fluorescent application of flow cytometry looking for an aneuploidy in cancer specimens. However, now-a-days the most common reason for measuring DNA content is because it gives us a very nice way to profile the position of cells within the cell cycle. Let's explore the cell cycle analysis plot in Kaluza Analysis Software together with Dr. David Onion.
The Composite feature in Kaluza Analysis will allow you to visualize multiple files in the same plot or report sheet, and enable you to create an overlay plot. Overlay plots are a powerful visual to present your data, allowing you to highlight small differences or demonstrate data sets to be very similar.
Radar and Tree plots allow you in Kaluza Analysis Software to visualize your data. Both of these plot types address the problem of understanding and visualizing the multi-dimensional data which is becoming ever more important as we begin to utilize more and more parameters as standard in our flow cytometry. They are particularly useful in dissecting multi-parameter phenotyping data and they can also help us to understand the relationship between multiple functional markers such as cytokines or phosphorylation events.
Consistency and accuracy of data is key to flow cytometry and so we often want to ensure this through some sort of quality control procedure. It might be routine QC of the machines through manufacture recommend controls or standards or you may wish to assess the reproducibility of a new staining panel that you are developing and so run a series of repeats on identical samples. Either way a Quality Control report gives you a quick way to analyze the data and spot any problems and outliers through the use of Levey-Jennings plots.