Phase contrast enhancement in microscopy using spiral phase filtering

We demonstrate two methods based on Fourier plane filtering using (a) a fractional spiral phase plate (SPP) and (b) an off-axial SPP for phase contrast enhancement in optical microscopy. In comparison to previous works, a spatially incoherent LED is used in the Kohler illumination as the light source to illuminate the biological specimen. We demonstrate that both these methods can transform the phase specimen into a relief-like view even under such illumination. The degree and orientation of enhancement can be controlled by changing the phase structure of the filter. The SPP is displayed on a phase-only spatial light modulator, and can be integrated into the optical path of standard microscopes. (C) 2009 Elsevier B.V. All rights reserved.