We present LIVE-PAINT, a new approach to super-resolution fluorescent imaging inside live cells. In LIVE-PAINT only a short peptide sequence is fused to the protein being studied, unlike conventional super-resolution methods, which rely on directly fusing the biomolecule of interest to a large fluorescent protein, organic fluorophore, or oligonucleotide. LIVE-PAINT works by observing the blinking of localized fluorescence as this peptide is reversibly bound by a protein that is fused to a fluorescent protein. We have demonstrated the effectiveness of LIVE-PAINT by imaging a number of different proteins inside live S. cerevisiae. Not only is LIVE-PAINT widely applicable, easily implemented, and the modifications minimally perturbing, but we also anticipate it will extend data acquisition times compared to those previously possible with methods that involve direct fusion to a fluorescent protein.
The first measurements of long wavelength (k y ρ s < 0.3) electron temperature fluctuations in Alcator C-Mod made with a new Correlation Electron Cyclotron Emission (CECE) diagnostic support a long-standing hypothesis regarding the confinement transition from Linear Ohmic Confinement (LOC) to Saturated Ohmic Confinement (SOC). Electron temperature fluctuations decrease significantly (∼40%) crossing from LOC to SOC, consistent with a change from Trapped Electron Mode (TEM) turbulence domination to Ion Temperature Gradient (ITG) turbulence as the density is increased. Linear stability analysis performed with the GYRO code [Candy and Waltz 2003 J. Comput. Phys. 186 545] shows that TEMs are dominant for long wavelength turbulence in the LOC regime and ITG modes are dominant in the SOC regime at the radial location (ρ ∼ 0.8) where the changes in electron temperature fluctuations are measured. In contrast, deeper in the core (ρ < 0.8), linear stability analysis indicates that ITG modes remain dominant across the LOC/SOC transition. This radial variation suggests that the robust global changes in confinement of energy and momentum occurring across the LOC/SOC transition are correlated to local changes in the dominant turbulent mode near the edge.
A correlation electron cyclotron emission (CECE) diagnostic has been installed in Alcator C-Mod. In order to measure electron temperature fluctuations, this diagnostic uses a spectral decorrelation technique. Constraints obtained with nonlinear gyrokinetic simulations guided the design of the optical system and receiver. The CECE diagnostic is designed to measure temperature fluctuations which have k(θ) ≤ 4.8 cm(-1) (k(θ)ρ(s) < 0.5) using a well-focused beam pattern. Because the CECE diagnostic is a dedicated turbulence diagnostic, the optical system is also flexible, which allows for various collimating lenses and antenna to be used. The system overview and the demonstration of its operability as designed are presented in this paper.
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