Las x version 3

The abundance and composition of MPs were investigated in a total of 40 samples of icefish (Neosalanx spp.) collected from several mass-market retailers in Calabria and Sicily regions in Italy. Samples were directly sourced as sales units: polyethylene (PE) plastic packages weighing between 250 – 1000 g of frozen, glazed and vacuumpacked icefish inside a paper case. As reported on the label, icefish of all the samples had been caught from the Taihu Lake (China), fished with trawls, gillnets or lift nets. Once collected, samples were brought to the laboratory and stored at -30 °C directly inside their original packaging until processing. The mouth size of 40 specimens randomly selected among the samples was observed with a stereomicroscope (Leica M205C, Germany) and measured Leica’s software (Leica LAS X version 3.0.14).

Filter of each sample and procedural blank were three times observed under a stereomicroscope (Leica M205C, Germany) by two different operators to record and categorized MPs and other anthropogenic items according to their shape, size and color (Hidalgo-Ruz et al., 2012 (link)). Stereomicroscope observation was performed outside the clean airflow cabinet; therefore, two blank filters were placed next to the samples during the analysis and checked for any occurred air-borne contamination. Any items found on the blank filters were morphologically compared with those detected in the filters of the samples. If too similar to each other, items found in the filter samples were not considered for the final count.
Based on their shape, the identified items were classified according to GESAMP (2019) in: i) “fragment” (MP-fr); ii) “foam” (MP-fo); iii) “film” (MP-fi); iv) “line” (MP-li); “pellet” (MP-pe) (Table 1). Leica’s software (Leica LAS X version 3.0.14) for the imaging analysis was used to measure the size of the items which were then classified into dimensional classes.

HuH-7 and HepG2 cells were plated at a density of 1.2×10⁵ cells/well in 12-well plates and allowed to adhere overnight. After ASHE treatment for 72 h, the cells were incubated with 75 nM DAPGreen (Dojindo Molecular Technologies, Inc.) and NucBlue Live ReadyProbes Reagent (Thermo Fisher Scientific, Inc.) at 37°C for 30 min. After washing with PBS, fluorescence imaging was performed using a Leica DMi8 fluorescence microscope (magnification, ×400) and LAS X version 3.3 (Leica Microsystems, Inc.).