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Stain Index Analysis

With FCS Express, you can easily create your own Stain Index to help you optimize your experiment for the best possible results. Background: In flow cytometry, experiment and panel design have become integral. As flow cytometry technology has evolved and more fluorochromes become available to researchers, a method to help optimize experimental design was needed. The Stain Index is a normalized signal to background metric that was first presented by David Parks of Stanford University at the International Society for Analytical Cytology annual meeting in Montpelier, France in 2004. The Stain Index was also featured in publications by Martin Bigos et al in 2004, then by Holden Maecker et al in 2008, with other modifications by other publishers in the following years. The Stain Index is a method to help determine the relative intensity of a fluorochrome on a given instrument and is calculated for a fluorescent parameter by subtracting the Median of the negative population from the Median of the positive population, then dividing that quantity by the variance of the negative population, multiplied by 2. Stain Index=(Median pos-Median neg)/(SD neg*2) By determining the relative brightness of a series of fluorochromes, a flow cytometry user can simplify the process by which they design an experiment and use this metric to match fluorochromes with a higher Stain Index to antigens that are of a lower density or a lower affinity to maximize resolution sensitivity. 00:00 Start 00:58 Outline of the Webinar 01:52 What is the Stain Index? 06:31 Calculating the Stain Index using Integrated Spreadsheets 16:52 Why are antibody titrations important? 18:30 Using the Universal Antibody Titration Template 36:20 Calculating the Stain Index using Custom Tokens 44:14 Summary 45:34 Additional Resources