In this paper, we demonstrate that it is possible to observe many aspects of critical phenomena in supported lipid bilayers using atomic force microscopy (AFM) with the aid of stable and precise temperature control. The regions of criticality were determined by accurately measuring and calculating phase diagrams for the 2 phase L d -L o region, and tracking how it moves with temperature, then increasing the sampling density around the estimated critical regions. Compositional fluctuations were observed above the critical temperature (T c ) and characterised using a spatial correlation function. From this analysis, the phase transition was found to be most closely described by the 2D Ising model, showing it is a critical transition. Below T c roughening of the domain boundaries occurred due to the reduction in line tension close to the critical point. Smaller scale density fluctuations were also detected just below T c . At T c , we believe we have observed fluctuations on length scales greater than 10 mm. The region of critically fluctuating 10-100 nm nanodomains has been found to extend a considerable distance above T c to temperatures within the biological range, and seem to be an ideal candidate for the actual structure of lipid rafts in cell membranes. Although evidence for this idea has recently emerged, this is the first direct evidence for nanoscale domains in the critical region.
Breaking the resolution limitUnder specific conditions; that is, with an atomically sharp tip and rapidly decaying tip-sample interaction forces, atomic resolution is attainable on flat incompressible materials such as mica by conventional AFM imaging 18 . Achieving and maintaining such conditions on biological samples, which are not only soft and dynamic, but also immersed in liquid at room temperature, is not possible. Typically, the tip geometry from the apex up to the height of the objects being imaged is much larger than the separation distance between the features of
Ribonuclease-A (RNase-A) encapsulated
PbS quantum dots (RNase-A@PbS
Qdots) which emit in the second near-infrared biological window (NIR-II, ca. 1000–1400 nm) are rapidly synthesized under microwave
heating. Photoluminescence (PL) spectra of the Qdots can be tuned
across the entire NIR-II range by simply controlling synthesis temperature.
The size and morphology of the Qdots are examined by transmission
electron microscopy (TEM), atomic force microscopy (AFM), and dynamic
light scattering (DLS). Quantum yield (Φf) measurement
confirms that the prepared Qdots are one of the brightest water-soluble
NIR-II emitters for in vivo imaging. Their high Φf (∼17.3%) and peak emission at ∼1300 nm ensure
deep optical penetration to muscle tissues (up to 1.5 cm) and excellent
imaging contrast at an extremely low threshold dose of ∼5.2
pmol (∼1 μg) per mouse. Importantly, this protein coated
Qdot displays no signs of toxicity toward model neuron, normal, and
cancer cells in vitro. In addition, the animal’s
metabolism results in thorough elimination of intravenously injected
Qdots from the body within several days via the reticuloendothelial
system (RES), which minimizes potential long-term toxicity in vivo from possible release of lead content. With a combination
of attractive properties of high brightness, robust photostability,
and excellent biocompatibility, this new NIR-II emitting Qdot is highly
promising in accurate disease screening and diagnostic applications.
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