Semrock

Why are these filter sets ideal for laser-based microscopy? Semrock’s laser fluorescence filter sets are crafted to take advantage of the superior resolution, higher sensitivity, and better image fidelity offered by today's state-of-the-art laser-based microscopes. They are optimized for the most popular lasers used in fluorescence imaging, including newer all-solid-state lasers that are rapidly replacing older gas-laser technology.

The excitation filters in these sets offer excellent transmission at the designated laser wavelengths, combined with low ripple passbands. The separation of the excitation and high-transmission emission filters is exceptionally small (less than 2.5% of the longest laser wavelength) to enable maximum capture of even weak signals close to the laser line. Matched BrightLine Laser Dichroic beamsplitters exhibit excellent reflection for all laser polarizations and transition rapidly to a wide and low-ripple fluorescence passband, with the flatness, ultra-low autofluorescence, and surface cosmetic properties needed for superior imaging performance.

Image registration performance: BrightLine Laser Fluorescence Filter Sets inherently provide excellent image registration performance – when interchanging these sets with one another, minimal pixel shift is observed. Note that the laser filter sets are not designed to exhibit “zero pixel shift” performance when interchanging with BrightLine ZERO™ filter sets. Images obtained with the laser filter sets do exhibit excellent image registration not only with one another, but also with images obtained when no fluorescence filter cube is present (e.g., DIC or other brightfield modes).

Applications include:

   • Any microscope or imaging system that benefits from premium beamsplitter spectral performance and/or flatness
   • Laser-scanning confocal microscopes (e.g. Olympus FluoView FV300, FV1000; Nikon Eclipse C1 Plus/si, A1)
   • Spinning-disk confocal microscopes (e.g. BD Biosciences CARV II; Olympus DSU)
   • Total-internal-reflection fluorescence (TIRF) microscopes (e.g. Olympus TIRF Illuminator, FV1000-EVA; Nikon TIRF)
   • Structured illumination microscopes (e.g. Qioptiq OptiGrid®)

Summary of available filter sets: The table below summarizes the most prominent lasers and fluorophores for which the current offerings of laser fluorescence filter sets have been optimized.

Prominent Laser Lines / Bands	Laser Description	Prominent Compatible Fluorophore(s)	Filter Set
375 ± 3 nm 405 ± 5 nm	GaN diode GaN diode	DAPI	LF405-A
473 ± 2 nm 488 +3/–2 nm 491 nm	Doubled DPSS^[1], Diode Ar-ion gas, Doubled OPSL^[2] Doubled DPSS^[1]	FITC, GFP	LF488-A
559 ± 5 nm 561.4 nm 568.2 nm	Diode Doubled DPSS^[1] Kr-ion gas	RFP’s (mCherry, HcRed, DsRed), Texas Red, TRITC, and Cy3	LF561-A
632.8 nm 635 +7/–0 nm 647.1 nm	HeNe gas AlGaInP diode, Kr-ion gas	Cy5	LF635-A

[1] DPSS = Diode-pumped solid-state laser
[2] OPSL = Optically pumped semiconductor laser