Background fluorescence includes any signal detected beyond what is generated by the fluorochromes being measured. The three major sources of background fluorescence include autofluorescence, spectral overlap, and undesirable antibody binding. Because false signals...
The field of flow cytometry continues to progress toward the goal of collecting and analyzing more data. Here we explore two recent advancements: spectral analyzer platforms to collect more data, and machine learning algorithms to analyze larger datasets. Machine...
Pathologists and life science researchers are increasingly performing multiplexed assays on formalin-fixed, paraffin-embedded samples. This allows for the collection of more data from a single tissue specimen, far beyond the typical single-color immunohistochemistry...
Compensation is the process of correcting for spillover when one fluorophore is detected in multiple channels. It is required for most experiments of four or more colors to identify the correct signal that should be measured in each channel. This quick list can help...
When designing a flow cytometry experiment, it is important to consider the relative brightness of each fluorescent label on your specific instrument. Generally, it is best to assign brighter fluorochromes to weakly expressed markers, and dimmer labels to strongly...
Flow cytometry can be performed directly using a fluorescently labeled primary antibody, or indirectly using an unconjugated primary antibody with a labeled secondary. Most researchers prefer direct detection due to its simplicity and ease of use. However, certain...
