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Methods of gene transfer to tissues are still to be optimized.
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Tumor oxygenation studies at microvascular levels can provide understanding of oxygen transport on scales at which oxygen transfer to tissue occurs.
Red blood cells were lysed with RBC lysis solution (Sigma), followed by repeated washing of extracted PBMCs with phosphate buffered saline and transfer to tissue culture plates pre-coated with poly-L-lysine (Sigma).
In the absence of effect studies on freshwater species, one can only speculate on potential sensitive endpoints: Ingested plastic fragments may most likely affect the metabolism (starvation due to decreased energy intake) and induce inflammation (when transferring to tissues).
However, it is also evident that elemental concentrations acquired in one season could be transferred to tissues grown in another season, highlighting the need to consider the carry-over effect of elemental concentrations between distinct oceanic areas.
Fresh and in vitro cultured trigeminal ganglia were transferred to Tissue-Tek (Sakura Finetek, USA), snap frozen on dry ice and stored at −80°C until further processing.
The excised tissues were washed thrice with PBS containing double strength antibiotics and the epithelial tissues were cut into multiple bits using sterile blade and the bits were transferred to tissue culture plates followed by incubation at 37 °C and 5%% CO2 for 5 min.
Embryos were transferred to tissue culture flasks and submerged in thermostatically controlled water baths to maintain a constant temperature of 28°C.
After two weeks, EBs were collected, transferred to tissue culture plates where they re-adhered, and ultimately passaged in fibroblast medium.
The stained tissues were transferred to tissue cassettes and paraffin blocks were prepared using standard methods.
After 9 days EBs were transferred to tissue culture dishes to attach.
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