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Overview
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Overview

The capabilities of flow cytometry instruments in terms of available lasers, detectors and number of measurable parameters have expanded tremendously over the years. Concomitant with that, there is an exponential increase in the number of fluorochromes available for flow cytometry, extending the limits of possibilities for new discoveries. Conventional flow cytometry collects only a discrete portion of the emission spectrum as defined by a single filter per fluorochrome, whereas spectral flow cytometry collects the full spectrum. This increases the flexibility and the capability of spectral flow by enabling autofluorescence profiling and extraction, detection of more fluorochromes per laser line and increased panel size. Find out how spectral flow cytometry differs from conventional flow cytometry and how to design your panels for spectral flow cytometry.
 

Read the blog article on how spectral flow cytometry has changed the landscape of fluorochrome development for high-dimensional research.

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Spectral vs conventional flow cytometry comparison
 

FeatureSpectralConventional
Detector and fluorochrome ratioMore detectors than fluorochromes1:1
Individual fluorochrome contribution determinationUnmixingCompensation
Fluorochrome choiceDependent on laser configurationDependent on laser and filter configuration
Autofluorescence extractionYesNo
Resolution of highly similar fluorochromesYesNo
Multi-color panel capability40+ colors~28 colors


What remains the same?

 

  • Workflow (sample prep, staining, acquisition)
  • Need for accurate single-stain controls for unmixing
  • Panel design principles

Introduction to spectral flow cytometry

Watch the spectral flow cytometry videos below to explore various topics, such as the fundamentals of spectral and conventional flow cytometry, how you can use more fluorochromes per laser line using spectral flow cytometry, how to use spectral flow cytometry for increased panel size, and more. 

Fundamentals of spectral versus conventional flow cytometry



The video addresses the following questions:

  • What is spectral flow cytometry?
  • What are the similarities and differences between conventional and spectral flow cytometry?
  • How do you read spectral signatures?
  • What is unique about spectral flow cytometry?
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      Autofluorescence unmixing
       

      The video addresses the following questions:

      •  What is autofluorescence unmixing?
      • How can autofluorescence impact biological resolution?
      •  What are cell-specific autofluorescence signatures?
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          Fluorochrome choice flexibility
           

          The video addresses the following questions:

          • How to select fluorochromes in either conventional or spectral flow cytometry?
          • How does spectral flow cytometry support the use of more fluorochromes per laser line?
          • What are the advantages of using spectral flow cytometry for fluorescent protein detection?

           

           

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              Increased panel size
               

              The video addresses the following questions:

              • How does spectral flow cytometry enable resolution of highly similar fluorochromes?
              • What are the similarity index and guidelines for the use of highly similar fluorochromes?
              • What are the specifics behind a 40-color multicolor spectral flow cytometry panel?
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                  Panel Design
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