Greetings Earthling,

Did you know...The mantis shrimp has SIXTEEN color-receptive cones which allow it to see a MASSIVE spectrum of color that human brains aren't even capable of processing (we only have 3 types of cones). Good thing the mantis shrimp will never get to design multicolor flow cytometry antibody panels... and good thing Boster has 6 helpful tips for humans to make multicolor panels right here! Click the shrimps for more FACS resources.

Mantis Shrimp

Troubleshooting Guides

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troubleshooting_box_image Troubleshooting guides

FACS antibodies multicolor panel design

  1. Always try to pair the brightest fluorochromes with the weakest expressing antigen. In case the expression level of an antigen is unknown, it is advisable to use brighter fluorochromes.
  2. Avoid spillover or spectral overlap between fluorochromes by spreading them as much as possible across the spectrum.
  3. Avoid fluorochromes that can be excited by more than one laser e.g. APC-Cy7.
  4. Always include suitable compensation controls (see our FACS resources for details) especially if points 3 and 4 above cannot be achieved.
  5. It is also advisable to use FMO controls (see our FACS resources for details) to gate differently stained sub-populations more accurately.
  6. Consider using online multicolor panel designers provided by FACS technology companies e.g. fluorofinder.com.

Example: A suitable 4-color panel to illustrate points 1-3. The color of the boxes correspond to the lasers that excite the respective fluorochromes.

These CD antibodies are available from Boster! Check recommended products below for validated FACS images...

example multicolor flow cytometry antibodies panel

Boster's Available Fluorophores (antibody conjugate labels):

Dyes Spectrum Graph

FACS troubleshooting ebook guide download PDF

*Note to educators: you are encouraged to share BosterBio's resources and PDFs on your class and lab websites, please cite or link to origin bosterbio.com

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