The difficulties identifying the underlying mechanisms in biomineralization are compounded by the lack of imaging techniques which simultaneously provide the desired chemical and elemental sensitivity to both identify sparse nanoscale organics and inorganic crystal structure across a large field of view. Here, we combine nanoscale secondary ion mass spectroscopy (nano-SIMS) and infrared scattering-scanning near-field optical microscopy (IR s-SNOM) to gain simultaneous elemental and chemical nano-images across 10’s of micrometer scales. We map the loss of order within the biomineralized material to gain insight in its formation processes whereby amorphous phases assemble into well-ordered macrostructures.
Extreme ultraviolet (EUV) lasers possess unique properties for ablation and ionization at the nanoscale (≤100 nm) due to their short wavelength, high absorptivity in most materials (i.e., 10’s of nanometers), and efficient photoionization in the laser-created plasmas. When coupled with a mass spectrometer, an EUV laser can be used to analyze and map chemical information in three dimensions with nanoscale spatial resolution. We have previously built an EUV time-of-flight mass spectrometer (EUV TOF) that achieved ~80 nm lateral and ~20 nm depth resolution when mapping the chemical content in organic and inorganic solids. Here, we present results from a recent study that extends EUV TOF’s high resolution capabilities to the analysis of an isotopically heterogenous uranium fuel pellet that was made by blending two isotopically distinct starting materials. We show that EUV TOF can map 235U/238U heterogeneity at the 100 nm scale, revealing micron to submicron heterogeneity. For comparison, nanoscale secondary ionization mass spectrometry (NanoSIMS) maps a similar distribution of U heterogeneity on a similar subsample at the same spatial scale. We also show that EUV TOF can measure the isotope ratio in a silver sample using single shot spectra. These results position EUV TOF as a promising technique for performing isotopic analyses at the nanoscale, finding applications in nuclear forensics, geology, and biology as well as in the semiconductor industry.
Several biotechnologies are currently available to quantify how cells allocate resources between growth and carbon storage, such as mass spectrometry. However, such biotechnologies require considerable amounts of cellular biomass to achieve adequate signal-to-noise ratio. In this way, existing biotechnologies inevitably operate in a ‘population averaging’ mode and, as such, they cannot unmask how cells allocate resources between growth and storage in a high-throughput fashion with single-cell, or subcellular resolution. This methodological limitation inhibits our fundamental understanding of the mechanisms underlying resource allocations between different cellular metabolic objectives. In turn, this knowledge gap also pertains to systems biology effects, such as cellular noise and the resulting cell-to-cell phenotypic heterogeneity, which could potentially lead to the emergence of distinct cellular subpopulations even in clonal cultures exposed to identical growth conditions. To address this knowledge gap, we applied a high-throughput quantitative phase imaging strategy. Using this strategy, we quantified the optical-phase of light transmitted through the cell cytosol and a specific cytosolic organelle, namely the lipid droplet (LD). With the aid of correlative secondary ion mass spectrometry (NanoSIMS) and transmission electron microscopy (TEM), we determined the protein content of different cytosolic organelles, thus enabling the conversion of the optical phase signal to the corresponding dry density and dry mass. The high-throughput imaging approach required only 2 μL of culture, yielding more than 1,000 single, live cell observations per tested experimental condition, with no further processing requirements, such as staining or chemical fixation.
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