Scepter software pro

Fatty-acid-based membrane lipidome analyses were performed by the Lipidomic Laboratory of Lipinutragen (Bologna, Italy). Blood samples (0.5 mL) collected in vacutainer tubes with ethylenediaminetetraacetic acid (EDTA) were treated according to the ISO17025 certified procedure (accredited Lab. #1836L) by robotic equipment and processed as described in previous studies [21 (link),22 (link),36 (link),46 (link)]. Briefly, the mature cell fraction was isolated based on the higher density of the aged cells with control of diameter controlled by cell counter (Scepter 2.0 with Scepter Software Pro, EMD Millipore, Darmstadt, Germany) [47 (link)]. After phospholipid extraction, derivatization to fatty acid methyl esters (FAME) was performed, transforming membrane glycerophospholipids (mainly phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidyl inositol, and plasmalogens) to examine up to 80% of the RBC membrane lipidome [48 (link)]. Fatty acids analysis was performed by gas chromatography (GC), and percentages are given as % relative quantitative (% rel. quant.), as previously described [21 (link),22 (link),36 (link),46 (link)], comparing with the benchmark of the interval values of each fatty acid and index [15 (link)].

Cellular concentration and size distribution of a log-phase culture were determined by using Scepter Handheld Automated Cell Counter (Merck Millipore), with 40 μm Scepter Cell Counter Sensors (Merck Millipore). 1 ml of yeast culture was sampled, sonicated, and diluted 1:100 in a final volume of 500 μl of PBS(1×). The diluted cells were vortexed and processed at the Cell Counter. Measured values were then exported with Scepter Software Pro (Merck Millipore). In Fig 5E, cellular concentrations are normalized with respect to the concentration at time 0.

Cell number and cell viability were measured using the automatic cell counter ADAM (NanoEnTek Inc., Seoul, Korea) according to the manufacturer’s recommendations. Cell diameters were measured using the Scepter 2.0 Cell Counter and Scepter Software Pro (Merck KGaA, Burlington, MA, USA) according to the manufacturer’s recommendations.