Autoquant x3

Manufactured by Oxford Instruments

Sourced in Switzerland

AutoQuant X3 is a X-ray microscopy instrument designed for high-resolution imaging and analysis of a wide range of sample types. It provides non-destructive, three-dimensional visualization and quantification of internal structures.

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Lab products found in correlation

Imaris 8, by Oxford Instruments (2 mentions) Huygens professional, by Scientific Volume Imaging (1 mentions)

3 protocols using autoquant x3

Image Deconvolution for Microscopy

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After acquisition, 3D stacks were processed for deconvolution. This image processing technique was performed to restore the imaged objects usually degraded by blurring and noise, artifacts inherent to any image acquisition system. The degree of blurring of a single, sub-resolution point-like object was considered as a measure of the optical system's quality and the blurred 3D image of this single point light source was defined by the Point Spread Function (PSF) [11] (link). Therefore, to counteract the effects of this blurring and to optimize image quality and subsequent quantifications, all images were processed using image processing software package (Huygens Professional, Scientific Volume Imaging b.v., Hilversum, The Netherlands) or image deconvolution software (Auto-Quant X3, Bitplane AG, Zurich, Switzerland).
Once images were processed to maximize quality, the relevant parameters were measured using image visualization and analysis software (Imaris 8.3.4, Bitplane AG, Zurich, Switzerland; Fiji ImageJ, Bethesda, Maryland, USA; IllucidaFX, Los Angeles, USA). Full descriptions of methodology and parameters studied are given in the subsequent sections.

Cribaro G.P., Saavedra E., Romarate L., Mitxitorena I., Díaz L.R., Casanova P.V., Roig-Martínez M., Gallego J.M., Pérez‐Vallés A., & Barcia C. (2020). Three-Dimensional Vascular Microenvironment Landscape in Human Glioblastoma.

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Deconvolution and Image Analysis for Enhanced Microscopy

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After acquisition, 3D stacks were processed for deconvolution. This image processing technique was performed to restore the imaged objects usually degraded by blurring and noise, artifacts inherent to any image acquisition system. The degree of blurring of a single, sub-resolution point-like object was considered as a measure of the optical system’s quality and the blurred 3D image of this single point light source was defined by the Point Spread Function (PSF) [11 (link)]. Therefore, to counteract the effects of this blurring and to optimize image quality and subsequent quantifications, all images were processed using image processing software package (Huygens Professional, Scientific Volume Imaging b.v., Hilversum, The Netherlands) or image deconvolution software (AutoQuant X3, Bitplane AG, Zurich, Switzerland).
Once images were processed to maximize quality, the relevant parameters were measured using image visualization and analysis software (Imaris 8.3.4, Bitplane AG, Zurich, Switzerland; Fiji ImageJ, Bethesda, Maryland, USA; IllucidaFX, Los Angeles, USA). Full descriptions of methodology and parameters studied are given in the subsequent sections.

Cribaro G.P., Saavedra-López E., Romarate L., Mitxitorena I., Díaz L.R., Casanova P.V., Roig-Martínez M., Gallego J.M., Perez-Vallés A, & Barcia C. (2021). Three-dimensional vascular microenvironment landscape in human glioblastoma. Acta Neuropathologica Communications, 9, 24.

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Deconvolution and Colocalisation Analysis

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Confocal images were deconvolved using Huygens Professional image processing software package (Scientific Volume Imaging b.v., Hilversum, The Netherlands) or AutoQuant X3 image deconvolution software (Bitplane AG, Zurich, Switzerland). Once images were processed to maximize quality, the colocalisation of the fluorescent materials was measured in voxels using Imaris 8.1 image visualization and analysis software (Bitplane AG, Zurich, Switzerland). Then, the colocalising voxels between the different fluorophores were extracted in a new channel to visualize the colocalisation. 3D isosurfaces were generated with the adequate detail resolution threshold and shadowing. Finally, 3D rotations were generated, and clipping planes were applied to show the position of cellular structures of interest, including the relevant colocalising material.

Conway G.E., He Z., Hutanu A.L., Cribaro G.P., Manaloto E., Casey A., Traynor D., Milosavljevic V., Howe O., Barcia C., Murray J.T., Cullen P.J, & Curtin J.F. (2019). Cold Atmospheric Plasma induces accumulation of lysosomes and caspase-independent cell death in U373MG glioblastoma multiforme cells. Scientific Reports, 9, 12891.

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