Imaging SPR combined with stereoscopic 3D tracking to study barnacle cyprid–surface interactions

Abstract

Barnacle larvae (cyprids) explore surfaces to identify suitable settlement sites. This process is selective, and cyprids respond to numerous surface cues. To better understand the settlement process, it is desirable to simultaneously monitor both the surface exploration behavior and any close interactions with the surface. Stereoscopic 3D tracking of the cyprids provides quantitative access to surface exploration and pre-settlement rituals. Imaging surface plasmon resonance (SPR) reveals any interactions with the surfaces, such as surface inspection during bipedal walking and deposition of temporary adhesives. We report on a combination of both techniques to bring together information on swimming behavior in the vicinity of the interface and physical interactions of the cyprid with the surface. The technical requirements for the setup are described, and we applied the setup to cyprids of Balanus amphitrite. Initial data shows the applicability of the combined setup to correlate exploration and touchdown events on surfaces with different chemical termination.

Resistance of Zwitterionic Peptide Monolayers to Biofouling

“…The settlement assays using settlement stage larvae of B. amphitrite show significant variations between surface coatings (Kruskal–Wallis, p < 0.002), and a Dunn’s test comparing the peptides show that that settlement preferences are similar for all peptides, in both assays, except for peptide 3 , for which settlement is nearly zero, and instead similar to that of the mPEG surface (Figure ). (Note: in this assay, only mPEG was used as a negative control, based on previous knowledge about its suitability for this purpose, and which is also demonstrated by our results.) To ensure that the absence of settlement onto peptide 3 is not caused by toxicity, extended settlement tests were performed to ensure that cyprids were able to at all settle on these SAMs (Figure S4, Supporting Information).…”

Biochemistry of Barnacle Adhesion: An Updated Review

“…and physicochemical properties, e.g., surface textures (Berntsson et al, 2000;Aldred et al, 2010;Chaw et al, 2011), hydrophobicity (Chaw and Birch, 2009;Phang et al, 2009;Guo et al, 2014), surface energy, and charge (Petrone et al, 2011;Di Fino et al, 2014) were also found to influence cyprid surface exploration. Using more complex video-/tracking-based approaches, the dynamic cyprid surface exploration process under different conditions was also monitored, offering a real-time and quantitative method of understanding the settling behaviors of a cyprid by measuring its swimming velocity, step length and duration, body movements, footprint deposition, and so on (Marechal et al, 2004;Andersson et al, 2009;Chaw and Birch, 2009;Maleschlijski et al, 2012;Aldred et al, 2013bAldred et al, , 2018Maleshlijski et al, 2016).…”

Quantitative analysis of the complete larval settlement process confirms Crisp's model of surface selectivity by barnacles

“…Scale bars: (c) 6 mm, (d) 10 mm, (e) 10 mm, (f) 2 mm, (g) 2 mm. [14,[16][17][18]20,21]. A significant advantage of our system is that it enables us to track multiple cyprids, simultaneously, throughout the entire pelagic-benthic transition to settlement, providing detail about the sequence of behaviours as well as the ability to compare between experimental conditions.…”

“…B 285: 20171957 individuals and, thus, how useful a behavioural assay may be as an indicator of propensity to settle. Previous studies have been limited to interpretation of short snapshots of behaviour out of context [14,[16][17][18]20,21]. A significant advantage of our system is that it enables us to track multiple cyprids, simultaneously, throughout the entire pelagic-benthic transition to settlement, providing detail about the sequence of behaviours as well as the ability to compare between experimental conditions.…”