Wide-field fluorescence sectioning with hybrid speckle and uniform-illumination microscopy.
We describe a method of obtaining optical sectioning with a standard wide-field fluorescence microscope. The method involves acquiring two images, one with nonuniform illumination (in our case, speckle) and another with uniform illumination (in our case, randomized speckle). An evaluation of the local contrast in the speckle-illumination image provides an optically sectioned image with low resolution. This is complemented with high-resolution information obtained from the uniform-illumination image. A fusion of both images leads to a full resolution image that is optically sectioned across all spatial frequencies. This hybrid illumination method is fast, robust, and generalizable to a variety of illumination and imaging configurations.
Lim
Daryl
D
Department of Biomedical Engineering, Boston University, Boston, MA 02215, USA.
Chu
Kengyeh K
KK
Mertz
Jerome
J
D003287:Contrast Media / Q000494:pharmacology; D004867:Equipment Design; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000494:pharmacology; D007091:Image Processing, Computer-Assisted / Q000379:methods; D008027:Light; D008856:Microscopy, Fluorescence / Q000379:methods*; D055095:Optics and Photonics; D011058:Pollen / Q000737:chemistry
D003287:Contrast Media; D005456:Fluorescent Dyes