<i>In situ vs ex situ</i> : comparing the structure of PNIPAM microgels at the air/water and air/solid interfaces

For studying the structure of microgel particles at the air/water interface, specular and off-specular X-ray reflectivity (XRR/OSR) allows <i>in situ</i> measurements without any labelling techniques. Herein we investigate the vertical and lateral structure of poly(<i>N</i>-isopropylacrylamide) (PNIPAM) microgels (MGs) at the air/water interface and the effect of Langmuir-Blodgett (LB) transfer onto solid substrates. The initial <i>ex situ</i> atomic force microscopy (AFM) scans of LB-transferred MGs at the air/solid interface reveal strong lateral 2D hexagonal ordering across a broad range of lateral surface pressures. Notably, for the first time, these results were confirmed by OSR, demonstrating the existence of the long-range hexagonal ordering at low and intermediate surface pressures. For <i>in situ</i> conditions and upon uniaxial compression at the air/water interface, the MG lattice constant decreases non-monotonically. This indicates the formation of domains at low pressures that approach each other and only compress when the surface isotherm reaches a plateau. Comparing the results of <i>in situ</i> and <i>ex situ</i> measurements, our study demonstrates a clear transfer effect during the LB-deposition on the lateral ordering of the MGs: the distance between the particles decreased during LB-transfer, and at high pressures (<i>Π</i> > 17 mN m^-1) a second distance occurs, indicating small domains with hexagonal internal ordering. The novel surface characterisation approaches debuted here highlight the use of both XRR and OSR to probe the vertical and lateral structure of adsorbed MGs, offering <i>in situ</i>, non-invasive insights without the need for doping or transfer-induced artefacts.