18 BD FACSCalibur Instructions for Use
Optics Components and Theory of Operation
The standard BD FACSCalibur system includes a 15-mW, 488-nm, argon-ion
laser and detectors for three fluorescent parameters. The FL4 option provides a
second 635-nm, red-diode laser and an additional detector. Laser light is focused
onto the flow cell by a focusing lens. As fluorescently tagged particles intercept
the laser light in the flow cell, scattered and fluorescent light provide information
about particle size, shape, granularity, and fluorescence intensity.
Forward scatter (FSC) signal is collected by the FSC diode. Side scatter (SSC) and
fluorescent signals are collected by a fluorescence collection lens and spectrally
split by a collection of dichroic mirrors (DMs) and filters. See page 220 for
wavelength characteristics of the dichroic mirrors and filters.
Figure 1-6 BD FACSCalibur optics system (standard)
SSC
FL1
FL2
FL3
488/10
530/30
90/10 beam splitter
DM 560SP
DM 640LP
585/42
650LP
FSC diode
488/10
focusing lens
collection lens
fluorescence
488-nm
blue laser
flow cell
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