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dc.contributor.authorSt John, James A
dc.contributor.authorKey, Brian
dc.date.accessioned2017-05-03T15:27:01Z
dc.date.available2017-05-03T15:27:01Z
dc.date.issued2012
dc.date.modified2014-08-28T22:16:46Z
dc.identifier.issn1567-2379
dc.identifier.doi10.1007/s10735-012-9462-7
dc.identifier.urihttp://hdl.handle.net/10072/47945
dc.description.abstractThe field of axon guidance is taking advantage of the powerful genetic and imaging tools that are now available to visualise growth behaviour in living cells, both in vivo and in real time. We have developed a method to visualise individual neurons within the living zebrafish embryo which provides exceptional cellular resolution of growth cones and their filopodia. We generated a DNA construct in which the HuC promoter drives expression of eGFP. Injection of the plasmid into single cell fertilised zebrafish egg resulted in mosaic expression of eGFP in neurons throughout the developing embryo. By manipulating the concentration of injected plasmid, it was possible to optimise the numbers of neurons that expressed the construct so that individual growth cones could be easily visualised. We then used time-lapse high magnification widefield epifluorescence microscopy to visualise the growth cones as they were exploring their environment. Growth cones both near the surface of the embryo as well as deep within the developing brain of embryos at 20 hours post fertilisation were clearly imaged. With time-lapse sequence imaging with intervals between frames as frequent as one second there was minimal loss of fluorescence intensity and the dynamic nature of the growth cones became evident. This method therefore provides high magnification, high resolution time-lapse imaging of living neurons in vivo and by use of widefield epifluorescence rather than confocal it is a relatively inexpensive microscopy method.
dc.description.peerreviewedYes
dc.description.publicationstatusYes
dc.format.extent717603 bytes
dc.format.mimetypeapplication/pdf
dc.languageEnglish
dc.language.isoeng
dc.publisherSpringer
dc.publisher.placeNetherlands
dc.relation.ispartofstudentpublicationN
dc.relation.ispartofpagefrom615
dc.relation.ispartofpageto623
dc.relation.ispartofissue6
dc.relation.ispartofjournalJournal of Molecular Histology
dc.relation.ispartofvolume43
dc.rights.retentionY
dc.subject.fieldofresearchBiochemistry and cell biology
dc.subject.fieldofresearchCardiovascular medicine and haematology
dc.subject.fieldofresearchClinical sciences
dc.subject.fieldofresearchCentral nervous system
dc.subject.fieldofresearchPeripheral nervous system
dc.subject.fieldofresearchcode3101
dc.subject.fieldofresearchcode3201
dc.subject.fieldofresearchcode3202
dc.subject.fieldofresearchcode320903
dc.subject.fieldofresearchcode320906
dc.titleHuC-eGFP mosaic labelling of neurons in zebrafish enables in vivo live cell imaging of growth cones
dc.typeJournal article
dc.type.descriptionC1 - Articles
dc.type.codeC - Journal Articles
gro.facultyGriffith Sciences, Griffith Institute for Drug Discovery
gro.rights.copyright© 2012 Springer Netherlands. This is an electronic version of an article published in Journal of Molecular Histology, December 2012, Volume 43, Issue 6, pp 615-623. Journal of Molecular Histology is available online at: http://link.springer.com/ with the open URL of your article.
gro.date.issued2012
gro.hasfulltextFull Text
gro.griffith.authorSt John, James A.
gro.griffith.authorEkberg, Jenny A.


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