Comparison of image deconvolution methods to improve image quality of 3D cell cultures acquired using HCS/HTS equipment

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dc.contributor.authorASSIS, JOAO V.A. de
dc.contributor.authorVIEIRA, DANIEL P.
dc.coverageInternacional
dc.creator.eventoLATIN-AMERICAN CONGRESS OF ARTIFICIAL ORGANS AND BIOMATERIALS, 12th
dc.date.accessioned2024-03-14T15:36:44Z
dc.date.available2024-03-14T15:36:44Z
dc.date.eventoDecember 12-15, 2023
dc.description.abstractIntroduction and objective: Accurate imaging analysis of 3D in vitro cell cultures became critical to evaluation of morphological and some physiological aspects of cells in these aggregates. Fluorescence microscopy protocols allow analysis of a plethora of molecular markers, DNA content and integrity and assessment of morphological changes upon different conditions. Light diffraction by plastic (or glass) substrate, and through sample and mounting/culture media distorts events, and thus images must require deconvolution methods to proper visualization and analysis. The work used 2 different algorithms and 3 iteration numbers to deconvolve images of spheroids acquired in large scale using HTS/HCS equipment. Methodology: Murine fibroblasts (NIH/3T3) were seeded on non-adhesive 96-well plates in RPMI 1640 medium. After 72 hours in culture, formed spheroids received pan-nuclear (Hoescht 33342) and nucleus of dead cells (SYTOX® Green) staining and imaged using HCS/HTS equipment (INCell 2500 HS, Cytiva). PSF files were generated using fluorescent beads in same cultured medium. Using DeconvolutionLab2 [1], a Fiji [2] plugin, spheroid images were deconvolved using the Tikhonov-Miller or Richardson-Lucy algorithms, with 10, 20 or 30 iterations. Orthogonal views of deconvolved stacks were analysed to find best results. Results and discussion: Both algorithms rendered similar results, with good reproducibility and resolution of light distortions. No real advantages were perceived in 30-iterations experiments, although 20 iterations rendered best images, regarding shape of nuclei (must be as spherical as possible) and drastic reduction of light refraction between slices. Conclusions: The work found that both methods were very easy to apply to stacks, with similar results. 20 iterations must be the better option to deconvolve spheroid images, uniting efficiency and saving computational resources. Richardson-Lucy can be the method of choice, as produced slightly better results.
dc.event.siglaCOLAOB
dc.format.extent215-215
dc.identifier.citationASSIS, JOAO V.A. de; VIEIRA, DANIEL P. Comparison of image deconvolution methods to improve image quality of 3D cell cultures acquired using HCS/HTS equipment. In: LATIN-AMERICAN CONGRESS OF ARTIFICIAL ORGANS AND BIOMATERIALS, 12th, December 12-15, 2023, Mar del Plata, Argentina. <b>Abstract...</b> p. 215-215. Disponível em: https://repositorio.ipen.br/handle/123456789/47929.
dc.identifier.orcidhttps://orcid.org/0000-0002-0007-534X
dc.identifier.urihttps://repositorio.ipen.br/handle/123456789/47929
dc.local.eventoMar del Plata, Argentina
dc.rightsopenAccess
dc.titleComparison of image deconvolution methods to improve image quality of 3D cell cultures acquired using HCS/HTS equipment
dc.typeResumo de eventos científicos
dspace.entity.typePublication
ipen.autorJOAO VICTOR ALMEIDA DE ASSIS
ipen.autorDANIEL PEREZ VIEIRA
ipen.codigoautor15846
ipen.codigoautor3158
ipen.contributor.ipenauthorJOAO VICTOR ALMEIDA DE ASSIS
ipen.contributor.ipenauthorDANIEL PEREZ VIEIRA
ipen.event.datapadronizada2023
ipen.identifier.ipendoc30277
ipen.notas.internasAbstract
ipen.type.genreResumo
relation.isAuthorOfPublication0ef49894-e71c-48f1-be08-2f29de9b4c1b
relation.isAuthorOfPublicationc2352608-be9c-4a73-be8c-571f10bb53d2
relation.isAuthorOfPublication.latestForDiscovery0ef49894-e71c-48f1-be08-2f29de9b4c1b
sigepi.autor.atividadeASSIS, JOAO V. A. DE:15846:810:N
sigepi.autor.atividadeVIEIRA, D.P.:3158:810:N
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