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Z-stack analysis and maximum intensity projections were performed using Volocity software (PerkinElmer, Waltham, MA, USA).
Deconvolution studies and 3D projections were performed using a DeltaVision system (Applied Precision, Issaquah, WA, USA) with an Olympus IX-71 microscope with × 100 objective/1.35 NA and × 60 objective/1.40 NA (Olympus, Tokyo, Japan) and filters set for DAPI, fluorescein isothiocyanate, and rhodamine provided by Applied Precision.
Deconvolution studies and 3D projections were performed using a DeltaVision system (Applied Precision, Issaquah, WA, USA) with an Olympus IX-71 microscope with × 100 objective/1.35 NA and × 60 objective/1.40 NA (Olympus, Shinjuku, Tokyo, Japan) and filters set for DAPI, fluorescein isothiocyanate and rhodamine provided by Applied Precision.
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Rotation-based forward and backward projections are performed using the PSF and attenuation factor [23].
Galerkin projection is performed using the extracted basis to yield a low dimensional I/O approximation of the original nonlinear model.
The regression step after projection is performed using computationally light models with low maintenance cost like Multiple Linear Regression and Locally Linear Regression with k-NN neighbourhoods.
The helical wheel projection was performed using the Gautier et al. ([2008]) algorithm present in the HeliQuest site (http://heliquest.ipmc.cnrs.fr) with a window size of 11 residues.
Tomographic reconstruction by Hamming-filtered back-projection was performed using NRecon software (Skyscan, Belgium).
Image deconvolution, projection, and colorization were performed using the SoftWoRx package and Photoshop CC 2014 (Adobe).
Indirect quantification of fluorescence levels, maximum intensity projections and kymograph construction were performed using ImageJ software [ 46].
Immediately after SPECT imaging was performed, CT images were acquired using the same coordinates as SPECT with 256 projections and 1024×1024 projection matrix size and a voltage of 60 kV; reconstructions were performed using filtered back-projections.
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