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BD FACSAria II cell sorter

Fluidics Sample to Sort Collection

Optics

Multicolor Controls and Analysis Services & Tools

Innovative optics design increases multicolor detection and efficiency.

Innovative designs for both the excitation optics and collection optics reduce excitation losses and dramatically improve collection efficiency.

The optical design allows for the use of lower powered, air cooled, and solid state lasers for efficiency without sacrificing sensitivity or resolution. No special power or cooling is needed.

Excitation optics

The excitation optics consist of lasers, fiber optic cables, beam-shaping prisms, and an achromatic focusing lens. Fiber optics precisely direct the laser light into beam-shaping prisms that transmit the laser light into a focusing lens. The lens focuses the laser light into the gel-coupled cuvette flow cell. Since the optical pathway and the sample core stream are fixed, alignment is constant from day to day.

Collection optics

The collection optics are set up in octagon- and trigon-shaped arrays that maximize signal detection from each laser. This is accomplished by transmitting the highest wavelengths to the first photomultiplier tube (PMT), and reflecting lower wavelengths to the next PMT through a series of longpass dichroic mirrors. Bandpass filters in front of each PMT allow fine-tuning of the spectral wavelengths that need to be collected.

This design is based on the principle that light reflection is more efficient than light transmission. Emitted light travels to each PMT detector via reflection and is transmitted through only two pieces of glass to reach the detector. This greatly increases sensitivity and resolution for each color in a multicolor panel.

Platform comparison using CD4

Comparison of whole blood stained with single-color CD4 FITC, CD4 APC, and CD4 Pacific Blue™ and run on both the BD FACSAria II and BD FACSCanto™ II systems. The BD FACSAria II was set for high-speed sorting (70 psi and 90 kHz) and both instruments were set up using BD Cytometer Setup and Tracking software.

Gel-coupled cuvette flow cell and fluorescence objective lens assembly

The cuvette is gel coupled to the fluorescence objective lens to transmit the greatest amount of emitted light from the interrogation point to the collection optics. The lens focuses fluorescent light emitted at each of the laser interrogation points onto individual collection fibers. These fibers transfer the emitted light to the collection optics.

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