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Main difficulties encountered in implementation of techniques for apoptosis detection in animal tissues

dc.contributor.authorGarcia, C.
dc.contributor.authorCruz, R.
dc.contributor.authorMega, A.
dc.contributor.authorOrtiz, A. L.
dc.contributor.authorTeixeira de Lemos, E.
dc.contributor.authorFerreira, D.
dc.contributor.authorReis, F.
dc.contributor.authorVala, Helena
dc.date.accessioned2014-04-11T13:01:23Z
dc.date.available2014-04-11T13:01:23Z
dc.date.issued2013
dc.description.abstractIntroduction: The implementation of methodologies capable of identifying apoptotic cells is a valuable tool in many biomedical studies. Much of the knowledge acquired about the mechanisms regulating apoptosis comes from observations in isolated cells; however, the properties shown by these assays do not always apply to the study of tissue samples. For detection of apoptosis in tissues, many investigators recommend the TUNEL method, nonetheless, other histochemical methodologies are available, and their choice must be carefully evaluated, taking into account the morphologic alterations which they intend to detect. Objective: To share encountered difficulties in the implementation of three different methods for the detection of apoptotic cells in animal tissue. Methods: Animal tissues, fixed in 10% neutral-buffered formalin and embedded in paraffin were submitted to immunohistochemical techniques using the TUNEL method (In situ Cell Death Kit, Roche) and polyclonal Bax and Bcl-2 antibodies (Santa Cruz Biotechnology). Results: After a long period of tests and costs, in which unspecific reactions, as well as several artifacts were obtained, the persistence to overcome these obstacles made all methods effective in the detection of apoptosis. Observed differences in immunolabeling were related to the type of tissues tested, which will be documented in the present study. Conclusions: Immunohistochemical methods are an approach to consider in the detection of apoptosis in animal tissues. Sharing the challenging difficulties found in our work, might obviate many tests and costs in its implementation in other laboratory facilities.por
dc.description.sponsorshipFCT and CI&DETS (PEst-OE/CED/UI4016)por
dc.identifier.citationGarcia C, Cruz R, Mega A, Ortiz AL, Teixeira de Lemos E, Ferreira D, Reis F, Vala H (2013). Main difficulties encountered in implementation of techniques for apoptosis detection in animal tissues. XVIII Meeting of the Portuguese Society of Animal Pathology. International Meeting of Leporidae Pathology, University of Évora. Portugal, 9-10 Maio.por
dc.identifier.urihttp://hdl.handle.net/10400.19/2070
dc.language.isoengpor
dc.peerreviewedyespor
dc.subjectapoptosispor
dc.subjectapoptotic eventspor
dc.subjectdetection of apoptosis in tissuespor
dc.subjectImmunohistochemical methodspor
dc.subjectTUNEL methodpor
dc.subjectBaxpor
dc.subjectBcl-2por
dc.titleMain difficulties encountered in implementation of techniques for apoptosis detection in animal tissuespor
dc.typeconference object
dspace.entity.typePublication
oaire.citation.conferencePlaceLisboapor
oaire.citation.titleRevista Portuguesa de Ciências Veterinárias. 108 (585-586): 77-78.por
person.familyNameVala Correia
person.givenNameHelena Maria
person.identifier.ciencia-id7A1E-E85E-FFA4
person.identifier.orcid0000-0001-6829-4867
rcaap.rightsrestrictedAccesspor
rcaap.typeconferenceObjectpor
relation.isAuthorOfPublicationcdc3d2e2-df06-40ed-8900-1ecbc8a06c8a
relation.isAuthorOfPublication.latestForDiscoverycdc3d2e2-df06-40ed-8900-1ecbc8a06c8a

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