The visualization data (8 movies) presented in this article are related to the research article entitled "Freezing and glass transitions upon cooling and warming and ice/freeze-concentration-solution morphology of emulsified aqueous citric acid" (A. Bogdan, M.J. Molina, H. Tenhu, 2016) [1]. The movies recorded in-situ with optical cryo-miscroscopy (OC-M) demonstrate for the first time freezing processes that occur during the cooling and subsequent warming of emulsified micrometer-scaled aqueous citric acid (CA) drops. The movies are made publicly available to enable critical or extended analyzes.

Calorimetric and optical cryo-microscope measurements of 10-64 wt % citric acid (CA) solutions subjected to moderate (3 K/min) and slow (0.5 and 0.1 K/min) cooling/warming rates and also to quenching/moderate warming between 320 and 133 K are presented. Depending on solution concentration and cooling rate, the obtained thermograms show one freezing event and from one to three liquid-glass transitions upon cooling and from one to six liquid-glass and reverse glass-liquid transitions, one or two freezing events, and one melting event upon warming of frozen/glassy CA/H2O. The multiple freezing events and glass transitions pertain to the mother CA/H2O solution itself and two freeze-concentrated solution regions, FCS1 and FCS2, of different concentrations. The FCS1 and FCS2 (or FCS22) are formed during the freezing of CA/H2O upon cooling and/or during the freezing upon warming of partly glassy or entirely glassy mother CA/H2O. The formation of two FCS1 and FCS22 regions during the freezing upon warming to our best knowledge has never been reported before. Using an optical cryo-microscope, we are able to observe the formation of a continuous ice framework (IF) and its morphology and reciprocal distribution of IF/(FCS1 + FCS2). Our results provide a new look at the freezing and glass transition behavior of aqueous solutions and can be used for the optimization of lyophilization and freezing of foods and biopharmaceutical formulations, among many other applications where freezing plays a crucial role.

The freezing of aqueous solutions and reciprocal distribution of ice and a freeze-concentrated solution (FCS) are poorly understood in spite of their importance in fields ranging from biotechnology and life sciences to geophysics and climate change. Using an optical cryo-microscope and differential scanning calorimetry, we demonstrate that upon cooling of citric acid and sucrose solutions a fast freezing process results in a continuous ice framework (IF) and two freeze-concentrated solution regions of different concentrations, FCS1 and FCS2. The FCS1 is maximally freeze-concentrated and interweaves with IF. The less concentrated FCS2 envelops the entire IF/FCS1. We find that upon further cooling, the FCS1 transforms to glass, whereas the slow freezing of FCS2 continues until it is terminated by a FCS2-glass transition. We observe the resumed slow freezing of FCS2 upon subsequent warming. The net thermal effect of the resumed freezing and a reverse glass-FCS1 transition produces the Ttr2-transition which before has only been observed upon warming of frozen hydrocarbon solutions and which nature has remained misunderstood for decades.