Focal pair merging for contrast enhancement of single particles.
A common technique in transmission electron microscopy is the collection of a focal pair, in which the first, close to focus image contains higher resolution information at lower contrast, and the second, far from focus image has high contrast but less reliable high-resolution information. Typically these second micrographs are used for purposes of particle selection, orientation estimate or micrograph evaluation. We introduce a technique for merging the information from both images, including signal to noise ratio weighting, contrast transfer function correction, and optional Weiner filtration. This produces a composite image with reduced contrast transfer function artifacts and optimized contrast. This technique is useful in numerous cases where low-contrast images are produced, such as small particles, proteins solubilized in detergent or projects with high-resolution goals when the first image is taken very close to focus.
Ludtke
Steven J
SJ
National Center for Macromolecular Imaging, Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX 77030, USA.
Chiu
Wah
W
D000465:Algorithms; D018834:Chaperonin 60 / Q000737:chemistry; D020285:Cryoelectron Microscopy / Q000379:methods*; D064429:Fatty Acid Synthases / Q000737:chemistry; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D008962:Models, Theoretical; D011506:Proteins / Q000737:chemistry
D018834:Chaperonin 60; D011506:Proteins; D064429:Fatty Acid Synthases