Shear free and blotless cryo-TEM imaging: a new method for probing early evolution of nanostructures

Cryogenic transmission electron microscopy (cryo-TEM) is a powerful method to image native state morphologies of nanoscale soft and hard objects suspended in solvents. Sample preparation is a critical step toward producing images at length and time scales of interest. We demonstrate a nearly shear-free sample thinning method which simultaneously allows imaging of evolving nanostructures at subsecond time scales. This device breaks the trade-off between high shear and short time scales typical in current cryo-TEM sample preparation methods. We demonstrate the low-shear feature of the new method by imaging wormlike micelles, showing an interconnected network, in contrast to the traditional sample preparation method which shows aligned micelles at similar time points. The time resolution of this method is demonstrated by imaging morphologies of calcium carbonate (formed through the reaction of calcium chloride with sodium carbonate) at subsecond time scales, capturing its evolution from an amorphous to a crystalline state. The impact of hyperbranched polyglycerol additives on the amorphous to crystalline transition in calcium carbonate at short times is examined. Early images at low shear provide unique fundamental insights into mechanisms of nanostructure evolution, thus offering a new paradigm for research in materials sciences, soft matter, and biological sciences.