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Sci Rep. 2014 Jun 5;4:5153. doi: 10.1038/srep05153.

Reassignment of scattered emission photons in multifocal multiphoton microscopy.

Author information

1
1] Massachusetts Institute of Technology, Department of Mechanical Engineering, Cambridge, MA 02139 [2].
2
1] Singapore-MIT Alliance for Research and Technology (SMART), BioSyM, Singapore 138602 [2].
3
Pohang University of Science and Technology, Department of Mechanical Engineering, Pohang 790-784, KOREA.
4
Massachusetts Institute of Technology, Picower Institute for Learning and Memory, Cambridge, MA 02139.
5
1] Institute of Bioengineering and Nanotechnology, A*Star, Singapore 138669 [2] Singapore-MIT Alliance, Computation and System Biology, Singapore 117576.
6
1] Singapore-MIT Alliance for Research and Technology (SMART), BioSyM, Singapore 138602 [2] Institute of Bioengineering and Nanotechnology, A*Star, Singapore 138669 [3] National University of Singapore, School of Medicine, Singapore 119077.
7
1] Massachusetts Institute of Technology, Picower Institute for Learning and Memory, Cambridge, MA 02139 [2] Massachusetts Institute of Technology, Departments of Biology, and Brain and Cognitive Sciences, Cambridge, MA 02139.
8
1] Massachusetts Institute of Technology, Department of Mechanical Engineering, Cambridge, MA 02139 [2] Singapore-MIT Alliance for Research and Technology (SMART), BioSyM, Singapore 138602 [3] Massachusetts Institute of Technology, Department of Biomedical Engineering, Cambridge, MA 02139.

Abstract

Multifocal multiphoton microscopy (MMM) achieves fast imaging by simultaneously scanning multiple foci across different regions of specimen. The use of imaging detectors in MMM, such as CCD or CMOS, results in degradation of image signal-to-noise-ratio (SNR) due to the scattering of emitted photons. SNR can be partly recovered using multianode photomultiplier tubes (MAPMT). In this design, however, emission photons scattered to neighbor anodes are encoded by the foci scan location resulting in ghost images. The crosstalk between different anodes is currently measured a priori, which is cumbersome as it depends specimen properties. Here, we present the photon reassignment method for MMM, established based on the maximum likelihood (ML) estimation, for quantification of crosstalk between the anodes of MAPMT without a priori measurement. The method provides the reassignment of the photons generated by the ghost images to the original spatial location thus increases the SNR of the final reconstructed image.

PMID:
24898470
PMCID:
PMC4046171
DOI:
10.1038/srep05153
[Indexed for MEDLINE]
Free PMC Article

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