Abstract
Despite
nanoparticles being used in many different products and applications, the
effects and fate in the environment are still not well understood. Uptake of
nanoparticles into cells has been shown in vitro and in vivo. However, it is
challenging to find suitable methods to identify uptake and determine
localization on a whole organism level. Furthermore, methods used to identify nanoparticle
uptake have been associated with artefacts induced by sample preparation
including staining methods for electron microscopy. This study used Fluorescent Light Sheet
Microscopy (FLSM) to determine uptake and localization of fluorescent labelled
nanoparticles in living whole organisms with minimal sample preparation. Two
strains of D. rerio (wildtype AB and transparent Casper) were exposed to
50 nm PEG coated gold nanoparticles (Au NP) synthesized with 1% of a fluorescent
probe (FITC). The fish were exposed to the particles through the diet or the
water phase in a series of separate experiments. In the dietary exposure
experiments Artemia salina were exposed to 1 mg Au/L for 24h before
being fed to D. rerio. For exposure through the water phase 1 mg Au/L
was added directly to aquaria holding the fish and non-exposed A. salina was
used for feeding. Imaging of D. rerio was done after 1, 3 and 7d of
uptake and again after 1 and 3d of depuration with FLSM for both dietary and
aqueous exposures. Though the FLSM proved to be excellent for in vivo detection
of NP, it was found that the opaqueness of wildtype AB zebrafish hindered deep penetration
of the light thus yielding less clear images compared to the Casper strain. A
time dependent increase in fluorescence was observed in the gut region of the
fish after dietary exposure. No fluorescence was observed outside the gut
channel indicating no or limited transfer of AuNP outside the gut region. Fish
exposed through the water phase showed adherence of AuNP to fins and especially
gills. Strong fluorescence signal was also observed in guts of fish due to
passive intake or drinking of exposed water. Fluorescence was also detectable
in parts of the head which could indicate uptake to the brain through the olfactory
tract. During the depuration phase the fluorescent signal decreased with time
for both exposure scenarios. These results suggest that the route of uptake is
pivotal for localization and transport of nanoparticles in zebrafish.
Furthermore, the study shows the suitability for whole imaging of living
organisms using FLSM.
Original language | English |
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Title of host publication | SETAC Europe 25th Annual Meeting : Abstract Book |
Number of pages | 1 |
Place of Publication | Barcelona, Spain |
Publisher | SETAC |
Publication date | 2015 |
Publication status | Published - 2015 |
Event | SETAC Europe 25th Annual Meeting - Barcelona, Spain Duration: 3 May 2015 → 7 May 2015 Conference number: 25 http://barcelona.setac.eu/?contentid=767 |
Conference
Conference | SETAC Europe 25th Annual Meeting |
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Number | 25 |
Country/Territory | Spain |
City | Barcelona |
Period | 03/05/2015 → 07/05/2015 |
Internet address |