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  Three-dimensional spectral imaging by Hadamard transform spectroscopy in a programmable array microscope

Hanley, Q. S., Verveer, P. J., Arndt-Jovin, D. J., & Jovin, T. M. (2000). Three-dimensional spectral imaging by Hadamard transform spectroscopy in a programmable array microscope. Journal of Microscopy, 197, 5-14.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0012-F866-F Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0027-E7CD-6
Genre: Journal Article

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 Creators:
Hanley, Q. S.1, Author              
Verveer, P. J.1, Author              
Arndt-Jovin, D. J.1, Author              
Jovin, T. M.1, Author              
Affiliations:
1Department of Molecular Biology, MPI for biophysical chemistry, Max Planck Society, ou_578628              

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Free keywords: 3-D imaging; deconvolution; Image reconstruction; Imaging spectroscopy
 Abstract: We report the acquisition and deconvolution of three-dimensional spectrally resolved images in a programmable array microscope implementing a Hadamard transform fluorescence spectroscopy system with adjustable spectral resolution. A stack of 16 two-dimensional spectral images was collected at 400 nm intervals along the optical axis. The specimen consisted of a polytene chromosome spread from Drosophila melanogaster doubly labelled for the Polyhomeotic protein by indirect immunofluorescence labelling with Alexa594 and for DNA with YOYO-1. The resulting four-dimensional data set consisted of the xyz spatial dimensions (898 x 255 x 16) with a 26-point spectrum at each spatial location. The total exposure time to the sample was 34 min. The system requires the acquisition of multiple images, and thus works best with fluorophores that are resistant to photobleaching. Image deconvolution reduced the amount of out-of-focus blur by up to a factor of 8, resulting in a dramatic improvement in the visualization of the chromosome backbone and localization of the specific Polyhomeotic domains.

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Language(s): eng - English
 Dates: 2005-08-172000
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: -
 Identifiers: eDoc: 226074
Other: 23123
 Degree: -

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Title: Journal of Microscopy
Source Genre: Journal
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Publ. Info: -
Pages: - Volume / Issue: 197 Sequence Number: - Start / End Page: 5 - 14 Identifier: -