Life Science Optics

Splitting Emission Signals

In fluorescence microscopy, there are several options available for viewing multiple colors from a single sample. The primary benefit of splitting the emission signals using dichroic beamsplitters is the ability to view multiple colors simultaneously rather than sequentially. This technique of using dichroics to separate emission signals is standard in flow instruments but requires specific arrangements when using fluorescence microscopes.

When the emission signals are being collected with a non-pixelated detector, such as a photomultiplier tube (PMT), any dichroic with appropriate transmission and blocking characteristics is acceptable. For signals being imaged with a pixelated detector, like a CCD, there are significant flatness requirements to consider.

Semrock's Image Splitting Dichroic Beamsplitters efficiently separate multicolored emission signals while maintaining excellent image fidelity. These dichroic beamsplitters, conveniently designed for 45° angle of incidence, are available for many popular fluorophore combinations. Their wide reflection & transmission bands and superb flatness allow for maximum light capture while minimizing image aberrations. Block excitation light and minimize bleedthrough with our vast selection of bandpass and long-pass emission filters.


White Papers

Flatness of Dichroic Beamsplitters Affects Focus and Image Quality

Optical Filters for Laser-based Fluorescence Microscopes

Related Publications

Achieving the Best Alignment for Fluorescent Images from the August 2005 issue of BioPhotonics

Fluorescence Imaging: Optical filters optimize laser-based fluorescence imaging systems from the January 2010 issue of Laser Focus World