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13 July 2004Reconstruction of objects with a limited number of non-zero components in fluorescence microscopy
A reconstruction algorithm is developed that uses
specific a-priori knowledge to produce higher
resolution images than standard approaches.
Deconvolution is an important image
reconstruction tool in fluorescence microscopy.
This is especially true for modern interferometric
instruments (such as I5M and 4Pi systems), as
they may have complicated oscillatory point
spread functions. Current methods are designed
to work on an arbitrary object - i.e. it is assumed
that there is no available a-priori knowledge of
the object (with the possible exception of a non-
negative condition on the fluorophore-emission
intensities). In situations where there is a-priori
knowledge of the object, it may be possible to use
this information to produce a higher quality
reconstruction of the object. A useful a-priori
condition is investigated here.
It is assumed that the object can be represented
by the sum of not more than L basis functions. The
simplest example of this is when the basis
functions are impulses - this leads to an object of
L or less non-zero points on a background of
zeros. This a-priori condition can be applied
directly; applied to a limited region of the object;
applied in one dimension (for an object with a
layered structure such as lipid bilayers); or
applied in two dimensions (for an object with a
filamentary structure such as actin fibers.) A
reconstruction algorithm is described and applied
to some illustrative simulated examples. The
results are found for several fluorescence
microscopy methodologies and compared to the
results produced by standard deconvolution
methods.
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Brynmor J. Davis, William C. Karl, Anna K. Swan, Bennett B. Goldberg, M. Selim Unlu, Marcia B. Goldberg M.D., "Reconstruction of objects with a limited number of non-zero components in fluorescence microscopy," Proc. SPIE 5324, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XI, (13 July 2004); https://doi.org/10.1117/12.527800