The observation of interference patterns in double-slit experiments with massive particles is generally regarded as the ultimate demonstration of the quantum nature of these objects. Such matter-wave interference has been observed for electrons 1 , neutrons 2 , atoms 3,4 and molecules [5][6][7] and it differs from classical wave-physics in that it can even be observed when single particles arrive at the detector one by one. The build-up of such patterns in experiments with electrons has been described as the "most beautiful experiment in physics" [8][9][10][11] . Here we show how a combination of nanofabrication and nanoimaging methods allows us to record the full two-dimensional build-up of quantum diffraction patterns in real-time for phthalocyanine molecules PcH2 and their tailored derivatives F24PcH2 with a mass of 1298 amu. A laser-controlled micro-evaporation source was used to produce a beam of molecules with the required intensity and coherence and the gratings were machined in 10-nm thick silicon nitride membranes to reduce the effect of van der Waals forces. Wide-field fluorescence microscopy was used to detect the position of each molecule with an accuracy of 10 nm and to reveal the build-up of a deterministic ensemble interference pattern from stochastically arriving and internally hot single molecules.When Richard Feynman described the double-slit experiment with electrons as "at the heart of quantum 2 physics" 12 he was emphasizing the fundamentally non-classical nature of the superposition principle which allows the quantum wave function associated with a massive object to be widely delocalized, while the object itself is always observed as a well-localized particle. Several recent experiments contributed to a further sharpening of the discussion by demonstrating the stochastic build-up of interferograms 11,13 , by implementing double-slit diffraction in the time-domain 14,15 , even down to the attosecond level 16 , and by identifying a single molecule as the smallest double-slit for electron interference 17,18 that enables fundamental decoherence studies 19 . The extension of far-field diffraction 20 to large molecules requires a sufficiently intense and coherent beam of slow and neutral molecules, a nanosized diffraction grating and a detector with both a spatial accuracy of a few nanometers and a molecule specific detection efficiency of close to 100 %. Our present experiment solves all these tasks simultaneously, using advanced micro-preparation, nanodiffraction and nanoimaging technologies. It thus exposes the quantum wave-particle duality in a particularly clear way and opens the way to new studies with ever larger molecules in an ongoing exploration of the quantumclassical borderline.Our setup is shown in Figure 1. It is divided into three parts: the beam preparation, coherent manipulation and detection. We need to prepare the molecules such that each of them interferes with itself and that all of them lead to similar interference patterns on the screen. Since the transverse and longitudina...