For the disease state to be characterized efficiently, choosing the correct fluorochromes for multiplexing cell phenotype markers and function in a single well is vital to resolving complicated cellular research questions.
Designing Multicolor Experiments is Simple
Know the instrument
- What are the number and type of lasers available?
- What detector or filter combinations are available for the signal to be captured?
This identifies what detection and labeling reagents are possible on users’ systems and how they will perform.
Know the detection reagents
- What are the emission spectra?
- What laser wavelength excites them?
- How bright are they? What is their stain index?
- Does spillover exist between adjacent channels?
- Beware that tandem and other dyes could degrade from exposure to temperature changes, light, or fixation.
These data will enable users to identify which fluorochrome best matches the marker of interest or antigen.
Know the biology
- The brightest fluorochromes could be reserved for the lowest expressed antigens and vice versa.
- Users can avoid spillover of highly expressed antigens into detectors that need high sensitivity.
Users have sufficient data to answer their experimental questions by improving fluorochromes and dyes.