What’s Your Objective?
Although optimizing the optical throughput of a fluorescence microscope requires attention to many factors including light source output, light source coupling to the microscope, optical filtering and camera quantum efficiency, objective selection remains most important of all.
For example, here is fluorescence image of a brine shrimp imaged with a 10X 0.5 NA objective. The two plots below the image are digital representations of the image data, showing the effect of switching to a 10X 0.3 NA objective with all other image acquisition parameters unchanged.
- Specimen: Eosin-stained brine shrimp
- SPECTRA X light engine®, single band exciter 470/24 (CWL/FWHM), 10% maximum output intensity. Semrock FF506-Di03 dichroic and FF01-545/55 emitter.
- Nikon Ti microscope, 10X, 0.5 NA objective OR 10X 0.3 NA objective, Hamamatsu ORCA-Flash4.0, 50 ms exposure.
Learn more about our SPECTRA X light engine used in the example above.
Download the PDF of Light BYTES: June 2016.