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Re‐evaluation of differential phase contrast (DPC) in a scanning laser microscope using a split detector as an alternative to differential interference contrast (DIC) optics
Author(s) -
Amos W. B.,
Reichelt S.,
Cattermole D. M.,
Laufer J.
Publication year - 2003
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1046/j.1365-2818.2003.01189.x
Subject(s) - differential interference contrast microscopy , optics , materials science , microscope , confocal , bright field microscopy , detector , interference (communication) , optical microscope , laser , microscopy , interference microscopy , birefringence , contrast (vision) , photodiode , numerical aperture , aperture (computer memory) , scanning electron microscope , physics , computer science , telecommunications , wavelength , channel (broadcasting) , acoustics
Summary In this paper, differential phase imaging (DPC) with transmitted light is implemented by adding a suitable detection system to a standard commercially available scanning confocal microscope. DPC, a long‐established method in scanning optical microscopy, depends on detecting the intensity difference between opposite halves or quadrants of a split photodiode detector placed in an aperture plane. Here, DPC is compared with scanned differential interference contrast (DIC) using a variety of biological specimens and objective lenses of high numerical aperture. While DPC and DIC images are generally similar, DPC seems to have a greater depth of field. DPC has several advantages over DIC. These include low cost (no polarizing or strain‐free optics are required), absence of a double scanning spot, electronically variable direction of shading and the ability to image specimens in plastic dishes where birefringence prevents the use of DIC. DPC is also here found to need 20 times less laser power at the specimen than DIC.