The BD LSR Fortessa is an advanced flow cytometer that uses multiple lasers and detection systems to analyze cells on various parameters, such as size, complexity, and fluorescence. When the BD LSR Fortessa is equipped with three lasers (blue, red, and violet), the system can simultaneously detect multiple fluorochromes (colors), allowing for detailed analyses with multiple markers in a single sample.

Here’s how the three lasers work:

1. Blue (B) – Wavelength: 488 nm

   • This is the most commonly used laser for flow cytometry. It excites fluorochromes such as:
     • FITC (Fluorescein isothiocyanate), PE (Phycoerythrin), PerCP, eFluor 450, etc.
     • The blue laser is ideal for fluorochromes that respond well to this wavelength.

2. Red (R) – Wavelength: 640 nm

   • The red laser excites fluorochromes such as:
     • APC (Allophycocyanin), Alexa Fluor 647, APC-Cy7, and Violet-conjugated fluorochromes.
     • It provides access to longer wavelengths that are required for certain fluorochromes that cannot be efficiently excited by the blue or violet lasers.

3. Violet (V) – Wavelength: 405 nm

   • The violet laser is used to excite fluorochromes such as:
     • Pacific Blue, V450, DAPI, V500, and other fluorochromes that respond to shorter wavelengths.
     • This laser is particularly useful for fluorochromes that cannot be efficiently excited by the blue or red lasers.

Advantages of 3 lasers in a BD LSR Fortessa:

With three lasers, you can use a broad range of fluorochromes simultaneously for multiparameter analysis. This enables the analysis of multiple cell markers in a single sample, which is ideal for applications such as:

• Immunophenotyping: Identifying different cell types based on marker expression.
• Cellular proliferation analysis: Measuring cell division and activation of specific signaling pathways.
• Apoptosis analysis: Studying cellular death processes using different fluorochromes for apoptosis markers.

By combining these three lasers, the BD LSR Fortessa can simultaneously collect data on various cellular parameters and marker expression with a high degree of precision, providing comprehensive and detailed analyses.