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Updated: Jun 24, 2025

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells
Published on: June 30, 2018
Self-referencing for quasi shot-noise-limited widefield transient microscopy
Abstract:
Many applications of ultrafast and nonlinear optical microscopy require the measurement of small differential signals over large fields-of-view. Widefield configurations drastically reduce the acquisition time; however, they suffer from the low frame rates of two-dimensional detectors, which limit the modulation frequency, making the measurement sensitive to excess laser noise. Here we introduce a self-referenced detection configuration for widefield differential imaging. Employing regions of the field of view with no differential signal as references, we cancel probe fluctuations and increase the signal-to-noise ratio by an order of magnitude reaching noise levels only a few percent above the shot noise limit. We anticipate broad applicability of our method to transient absorption, stimulated Raman scattering and photothermal-infrared microscopies.
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