ABSTRACT
Bioluminescence from cells is so dim that bioluminescence microscopy is performed using an ultra low-light imaging camera. Although the image sensor of such cameras has been greatly improved over time, such improvements have not been made commercially available for microscopes until now. Here, we customized the optical system of a microscope for bioluminescence imaging. As a result, bioluminescence images of cells could be captured with a conventional objective lens and colour imaging camera. As bioluminescence microscopy requires no excitation light, it lacks the photo-toxicity associated with fluorescence imaging and permits the long-term, nonlethal observation of living cells. Thus, bioluminescence microscopy would be a powerful tool in cellular biology that complements fluorescence microscopy.
Subject(s)
Microscopy, Fluorescence/instrumentation , Microscopy, Fluorescence/methods , Cell Line, Tumor , Firefly Luciferin , Fluorescent Dyes , Humans , Luciferases, Firefly , Luminescent MeasurementsABSTRACT
A beam-deflection type laser scanning microscope for differential phase contrast images using the split-detector technique is developed and the images are obtained in real time (30 images/s). The special optical design allows transfer of the pupil information necessary to the imaging of the differential phase contrast images during beam deflection and the use of acousto-optic deflectors with the associated correction system enables real time imaging. Differential phase images of surface details of crystals are simply obtained without silver coating or etching, which are indispensable in a normal DIC. Biological specimens are also observed.
