SBA-15 is an hexagonal mesoporous material which is synthesized with nonionic poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) block copolymers (Pluronics, EOyPOxEOy), templates. Pore diameters in the range of 2-30 nm can be obtained with a relatively thick silica wall (up to 6 nm). This material possesses both large, uniform, and ordered channels, along with a complementary net of micropores which provides connectivity between the ordered channels through the silica. This study focuses on the investigation of the formation mechanism of SBA-15 with emphasis on the PEO interactions with the silica and the initiation of the micropores. This was achieved using in situ X-band EPR spectroscopy in combination with electron spin-echo envelope modulation (ESEEM) experiments. The paramagnetic centers were introduced as spin-labeled Pluronic L62 (EO6PO30EO6) where nitroxides replace the OH groups at the end of the polypropylene oxide (PEO) blocks (L62-NO). Initially, the acidic reaction conditions were adjusted to prevent the decomposition of the nitroxide radical, while still producing highly ordered SBA-15. Then, the locations of the nitroxides of L62-NO within the micelles of Pluronic P123 (y = 20, x = 70) and L64 (y = 13, x = 30) were determined through three-pulse ESEEM experiments on solutions prepared in D2O. In these experiments, the H-2 modulation induced by D2O was compared with that of a series of small spin-probes with known hydrophilic and hydrophobic characters that were introduced into the micelles. The NO group of L62-NO was found to be close to the core-corona interface in both types of Pluronics. The temporal evolution of the EPR spectrum during the reaction showed that for SBA-15 made with P123 the most significant changes in the L62-NO spectrum occur within the first 100 min. Furthermore, X-ray diffraction measurements of dried materials showed that the hexagonal structure of SBA-15 is also created within the first 2 h. A partitioning of the L62-NO between the precursors of the mesopores and micropores of the SBA-15 structure takes place at the very early stages of the reaction, and a continuous depletion of water within the corona-core interface was observed. In the final product obtained without a thermal stage, the majority of the PEO chains are located in the micropores. The extent of the PEO chains located within the silica micropores depends on the thermal stage temperature and on the Si/P123 molar ratio. In the L64 synthesis, practically all of the NO groups of L62-NO are located within the silica network and experience a single environment.