Conventional and spectral flow cytometry are based on the same fundamental principles and even use some common hardware. However, spectral flow cytometry is a more advanced technique and offers several advantages over the conventional technique.

Here’s a look at the differences between conventional and spectral flow cytometry.

Conventional flow cytometry:

• Employs mirrors and optical filters to reflect, block, or transmit a photon based on its wavelength.
• Uses a one detector – one fluorophore setup where each detector is assigned to a specific fluorophore.
• Detectors used are individual photodiode (PD) or photomultiplier tube (PMT).
• Only a fraction of the fluorophore emission spectra is captured and recorded.
• Is capable of detecting signals from specific fluorophore over defined wavelengths.
• Fluorophores with similar emission spectra are different to distinguish.
• Autofluorescence can hamper data analysis, particularly in larger and more granular cells. 

Spectral flow cytometry:

• Employs dispersive optics such as spectrographs or prisms to disperse photons according to wavelength across an array of detectors.
• All detectors in the cytometer capture a signature of the full emission spectra of each fluorophore.
• Uses a linear array of detectors to measure the continuous spectrum generated by dispersive elements. Common detectors used are arrays of charge coupled devices (CCDs) and multianode PMTs
• Spectral flow cytometers are capable of detecting multi-parametric panels with more than 30 different fluorophores.
• Is capable of collecting the entire spectral profile of fluorophores from multiple lasers. 
• Fluorophores with similar emission spectra can be easily distinguished.
• Autofluorescence is treated as another color and can be removed through spectral unmixing and analysis, allowing for clearer identification of low-expressing targets and dim fluorescence signals.