Nano-carriers have the potential to be an enormous game-changer in medicinal drug delivery systems. The polymeric nano-carriers used in this study are a product of the self-assembly of amphiphilic block copolymers, a complicated process which must be understood completely to finely tune the desired morphology for drug delivery. The goal of this thesis is to gain a better understanding of the self-assembly process of amphiphilic block copolymers. Specifically, it will focus on the ’opaque phase’ observed for poly(1,2-butadiene)-b-poly(ethylene oxide) (PBd-PEO) block copolymers, which seems to occur in the early stages of the self-assembly process. A nano-precipitation method has been developed at the TU Delft, which induces selfassembly and brings forward the opaque phase. The used block copolymer has a hydrophobic PBd block and a hydrophilic PEO block. This block copolymer dissolves well in acetone, but upon water (H2O) addition, it starts to self-assemble into spherical aggregates, useful for drug delivery. At small volumes of H2O, the opaque phase appears and disappears as more H2O is added. In this thesis, multiple samples have been prepared with the so-called Inverse Nanoprecipitation method and different experimental parameters among which the volume percentage of H2O present in the sample, have been varied. The samples have been studied using Visual Inspection, Dynamic Light Scattering and Spin Echo Small Angle Neutron Scattering. The experiments show that the time intervals between H2O addition do not affect the formation of aggregates, but rather the ‘when’ of adding the H2O. If this is added to the acetone before the block copolymer is dissolved, it affects the self-assembly process. A visual experiment showed that the opaque phase occurred 1.2±0.1 vol% H2O earlier than in previous research, which might be a result of the lower room temperature during this thesis. Another significant result might be that the addition of acetone-D6 or D2O affects the self-assembly process, which must be considered for future SESANS measurements. Lastly, during the opaque phase a strong temperature sensitivity is observed (which was already found in previous research at TU Delft, by E. Remmelts and further researched by R. Baaijens), high light scattering intensities are detected with DLS and for SESANS measurements the scattered neutron intensities were low. These observations all strongly point to a theory called ‘pre-micellization’, which gives a better understanding of the opaque phase.