Total Fluorescence Yield

Total fluorescence yield is an X-ray absorption spectroscopy detection mode that measures the combined intensity of fluorescence emitted after a sample absorbs incident X-rays. It matters because the signal provides an element-selective measure of absorption while integrating emission from all relevant fluorescence lines. When an X-ray ejects a core electron, the resulting vacancy is filled by an electron from a higher energy level, releasing energy as characteristic fluorescence; a detector records these photons to generate an absorption spectrum. In chemistry, total fluorescence yield supports XANES and EXAFS studies of oxidation state, coordination environment, and elemental composition, especially in bulk or dilute materials.

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Education

JoVE Core - Biology

ATP Yield

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2019

Cellular respiration produces 30 - 32 ATP per glucose molecule. Although most of the ATP results from oxidative phosphorylation and the electron transport chain (ETC), 4 ATP are gained beforehand (2 from glycolysis and 2 from the citric acid cycle). The ETC is embedded in the inner mitochondrial membrane and is comprised of four main protein complexes and an ATP synthase. NADH and FADH2 pass electrons to these complexes, which pump protons into the intermembrane space. This distribution of...

Total Internal Reflection Fluorescence Microscopy

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2023

Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.

Research

JoVE EoE - Neuroimaging

Cell Imaging Using Total Internal Reflection Fluorescence Microscopy

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2025

Source: Daniele, F., et. al TIRFM and pH-sensitive GFP-probes to Evaluate Neurotransmitter Vesicle Dynamics in SH-SY5Y Neuroblastoma Cells: Cell Imaging and Data Analysis. J. Vis. Exp. (2015).This video demonstrates the imaging of human neuroblastoma cells using a total internal reflection fluorescence (TIRF) microscope to visualize fluorophore-tagged synaptic vesicles. The cells are first focused in epifluorescence mode. Adjustments are then made to achieve total internal reflection,...

Total Internal Reflection Fluorescence Microscopy for Visualization of Exocytic Events

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2025

Source: Winkle, C. C., et al. Utilizing Combined Methodologies to Define the Role of Plasma Membrane Delivery During Axon Branching and Neuronal Morphogenesis. J. Vis. Exp. (2016)This video demonstrates total internal reflection fluorescence (TIRF) microscopy for visualizing exocytic events in cortical neurons. A pH-sensitive green fluorescent protein (GFP) marker fluoresces upon vesicle fusion, with selective TIRF excitation capturing membrane-proximal exocytosis in time-lapse imaging.

Reaction Yield

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2020

The theoretical yield of a reaction is the amount of product estimated to form based on the stoichiometry of the balanced chemical equation. The theoretical yield assumes the complete conversion of the limiting reactant into the desired product. The amount of product that is obtained by performing the reaction is called the actual yield, and it may be less than or (very rarely) equal to the theoretical yield. Percent Yield In the case of chemical reactions, the actual yield of the product is...

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