All padded samples were dried at 100°C for 3 min, cured at 150°C for 3 min and finally rinsed with warm water (40°C) for 1 min.
After removal from the sweat pad, the sweat samples were frozen at −20°C in 18 ml batches (one per experiment) to avoid multiple thaws.
Exponentially growing cells were placed on agarose pads, and fresh samples were prepared every 15 min for all microscopy experiments.
Epididymal fat pads and blood samples were harvested immediately after euthanization, weighed, flash frozen in liquid nitrogen, and stored at −80 °C.
The monoexponential decays were analysed by using Graph Pad Prism 5. Samples were held in a 10×10 nm or 10×4 nm quartz Hellma cuvette and cutoff filters were used to avoid second-order diffraction effects.
The pads with oral fluid samples were transported to the laboratory at the Instituto Nacional de Saúde every day.
The weird hardcore pads and samples are almost Terminator-ish, but the structure holds it all together into something more functional.
Plasma concentrations of glucose were determined using diagnostic kits (ABX, Shefford, UK) on a Cobas Mira Plus automatic analyser (ABX .Liver, white adipose tissue (retroperitoneal fat pad) and skeletal muscle (soleus) samples were obtained from each of the 15 rats.
In the beginning of the session, the cotton pads were fixed, and saliva samples were obtained at this point (baseline), after the first break (30 min later, t1), after the third bicycling set (60 min later, t2), and at the end of the session (90 min later, t3).
The samples were sent in padded envelopes by regular mail to SVA where they usually arrived within 24 hours of sampling.
