title: Beyond the transect: an alternative microchemical imaging method for fine scale analysis of trace elements in fish otoliths during early life.
abstract: Microchemical analysis of otolith (calcified 'ear stones' used for balance and orientation) of fishes is an important tool for studying their environmental history and management. However, the spatial resolution achieved is often too coarse to examine short-term events occurring in early life. Current methods rely on single points or transects across the otolith surface, which may provide a limited view of elemental distributions, a matter that has not previously been investigated. Imaging by laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) permits microchemical analyses of short-term events in early life with high (<10 μm) resolution, two-dimensional (2D) visualization of elemental distributions. To demonstrate the potential of this method, we mapped the concentrations of Sr and Ba, two key trace elements, in a small number of juvenile otoliths of neon damselfish (Pomacentrus coelestis) using an 8 μm beam diameter (laser fluence of 13.8 ± 3.5 Jcm(-2)). Quantification was performed using the established method by Longerich et al. (1996), which is applied to 2D imaging of a biological matrix here for the first time. Accuracy of >97% was achieved using a multi-point non matrix-matched calibration of National Institute of Standards and Technology (NIST) 610 and 612 (trace elements in glass) using Longerich's calculation method against the matrix-matched standard FEBS-1 (powdered red snapper [Lutjanus campechanus] otolith). The spatial resolution achieved in the otolith corresponded to a time period of 2 ± 1 days during the larval phase, and 4 ± 1 days during the post-settlement juvenile phase. This method has the potential to improve interpretations of early life-history events at scales corresponding to specific events. While the images showed gradients in Sr and Ba across the larval settlement zone more clearly than single transects, the method proved sample homogeneity throughout the structure; demonstrating that 2D scanning has no significant advantage over line scans.
authors.affiliation: Elemental Bio-imaging Facility, University of Technology, Sydney, New South Wales, Australia; CCFS ARC Centre of Excellence, Department of Earth and Planetary Sciences, Macquarie University, Sydney, Australia.
authors.forename: Nicole
authors.initials: N
authors.lastname: McGowan
authors.affiliation: Fish Ecology Laboratory, School of the Environment, University of Technology, Sydney, New South Wales, Australia.
authors.forename: Ashley M
authors.initials: AM
authors.lastname: Fowler
authors.affiliation: Fish Ecology Laboratory, School of the Environment, University of Technology, Sydney, New South Wales, Australia.
authors.forename: Kerryn
authors.initials: K
authors.lastname: Parkinson
authors.affiliation: Elemental Bio-imaging Facility, University of Technology, Sydney, New South Wales, Australia.
authors.forename: David P
authors.initials: DP
authors.lastname: Bishop
authors.affiliation: Florey Department of Neuroscience and Mental Health, University of Melbourne, Victoria, Australia.
authors.forename: Katherine
authors.initials: K
authors.lastname: Ganio
authors.affiliation: Elemental Bio-imaging Facility, University of Technology, Sydney, New South Wales, Australia.
authors.forename: Philip A
authors.initials: PA
authors.lastname: Doble
authors.affiliation: Fish Ecology Laboratory, School of the Environment, University of Technology, Sydney, New South Wales, Australia.
authors.forename: David J
authors.initials: DJ
authors.lastname: Booth
authors.affiliation: Elemental Bio-imaging Facility, University of Technology, Sydney, New South Wales, Australia; Florey Department of Neuroscience and Mental Health, University of Melbourne, Victoria, Australia. Electronic address: dominic.hare@uts.edu.au.
authors.forename: Dominic J
authors.initials: DJ
authors.lastname: Hare
mesh_terms: D000818:Animals; D004784:Environmental Monitoring / Q000379:methods*; D005399:Fishes / Q000378:metabolism*; D013058:Mass Spectrometry; D010037:Otolithic Membrane / Q000737:chemistry / Q000378:metabolism*; D014131:Trace Elements / Q000032:analysis* / Q000378:metabolism; D014874:Water Pollutants, Chemical / Q000032:analysis / Q000378:metabolism*
chemical_list: D014131:Trace Elements; D014874:Water Pollutants, Chemical
keywords: Barium; Elemental imaging; LA-ICP-MS; Otolith; Reef fish; Strontium