. Author manuscript; available in PMC: 2020 Jul 18.

Published in final edited form as: Methods Enzymol. 2019 Jul 18;628:191–221. doi: 10.1016/bs.mie.2019.06.016

Table 3.

Typical components that might be used to construct a fluorescence detection pathway. Example specifications are designed for fluorescein-tagged compounds with an excitation wavelength near 488 nm and emission wavelength near 525 nm.

Component name	Selection considerations	Example specifications

Excitation Laser	Single wavelength, low power, continuous wave	Wavelength-488 nm
Excitation Laser	Single wavelength, low power, continuous wave	Power-20 mW

Single mode fiber optic	Consistent with laser wavelength	Operating wavelength-405–532 nm

Dichroic mirror	Reflect excitation light and pass emission light	Transmission band-520–800 nm
		Reflection band-380–490 nm
		Cut-on wavelength-505 nm

Microscope objective	High NA and compatible with filter cube	Magnification-40x
Microscope objective	High NA and compatible with filter cube	NA-0.65

Multi-mode fiber optic	Consistent with emission wavelength and large core diameter	Core diameter-25 ± 3 μm
Multi-mode fiber optic	Consistent with emission wavelength and large core diameter	Wavelength range-400–550 nm

Photo multiplier tube (PMT)	Large dynamic range and high sensitivity	Wavelength range-300–720 nm
Photo multiplier tube (PMT)	Large dynamic range and high sensitivity	Sensitivity-176 mA/W

Fiber optic couplers	Adjustable in three dimensions and low hysteresis

Collimator	Couples directly to fiber optic

Filter cube	Compatible with dichroic, couplers, objective, and collimator