Separation by TLC of the glycerolipids from P. tricornutum. Lipids from Pt1 cells grown in a replete medium (10N10P) were extracted and resolved following the procedures described in “Materials and Methods.” The cross indicates the initial deposit. A, One-dimensional separation of nonpolar lipids (DAG and TAG) and free FA (FFA). Migration was performed in hexane:diethylether:acetic acid (70:30:1, v/v). B, Two-dimensional separation of polar (membrane) lipids. Migration was performed in chloroform:methanol:water (65:25:4, v/v) for the first dimension (arrow 1) and chloroform:acetone:methanol:acetic acid:water (50:20:10:10:5, v/v) for the second migration (arrow 2). Lipids were visualized under UV light, after spraying with 2% 8-anilino-1-naphthalenesulfonic acid in methanol, and scraped off the plate for analyses. Identification of the lipid in each spot was performed by MS2 analyses. The spot circled in white is an unknown compound with a structure that differs from a glycerolipid.