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Related Concept Videos

Diffusion01:12

Diffusion

196.6K
Diffusion is the passive movement of substances down their concentration gradients—requiring no expenditure of cellular energy. Substances, such as molecules or ions, diffuse from an area of high concentration to an area of low concentration in the cytosol or across membranes. Eventually, the concentration will even out, with the substance moving randomly but causing no net change in concentration. Such a state is called dynamic equilibrium, which is essential for maintaining overall...
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Drug Absorption Mechanism: Passive Membrane Transport01:23

Drug Absorption Mechanism: Passive Membrane Transport

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Passive transport is a method of drug absorption where small, lipid-soluble drugs can move across the cell membrane. This movement happens along the concentration gradient, which is a natural flow from higher to lower concentration areas. The speed at which the drug moves is directly related to its lipid–water partition coefficient. This means that the more a drug dissolves in lipids, the faster it diffuses or spreads throughout the body. It is important to note that most drugs are either...
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Passive Diffusion: Overview and Kinetics01:17

Passive Diffusion: Overview and Kinetics

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Passive diffusion is a critical process that allows small lipophilic drugs to cross the cell membrane along a concentration gradient. This mechanism's efficiency depends on four primary factors: the membrane's surface area, the drug's lipid-water partition coefficient, the concentration gradient, and the membrane's thickness.
When administered orally, drugs establish a substantial concentration gradient between the gastrointestinal (GI) lumen and the bloodstream, expediting...
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The Significance of Membrane Transport01:44

The Significance of Membrane Transport

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The transport of solutes across the cell membrane is essential for metabolic processes, like maintaining cell size and volume, generating the action potential, exchanging nutrients and gases, etc. Membrane transport can be either passive or active. It can be simple diffusion, facilitated, or mediated transport aided by transport proteins such as transporters and channels.
Transporters facilitate either an active or passive movement of solutes. They can allow a single-molecule transport down its...
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Mechanisms of Drug Absorption: Paracellular, Transcellular, and Vesicular Transport01:23

Mechanisms of Drug Absorption: Paracellular, Transcellular, and Vesicular Transport

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Drugs need to permeate cell membranes to reach their target sites after administration. Orally administered drugs must transcend intestinal epithelial membrane barriers to infiltrate the systemic circulation. Drugs with a molecular weight of less than 500 Daltons diffuse through gaps between neighboring cells, called paracellular pathways.
However, most drugs use the transcellular route, traversing directly through the cell membranes via two mechanisms: passive and active transport. Passive...
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Cellular Membranes and Drug Transport01:24

Cellular Membranes and Drug Transport

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Drugs must traverse multiple biological barriers, such as multi-layered skin, single-layered intestinal epithelium, and the plasma membrane, to reach their target sites within the body. The plasma membrane, a highly structured composite of phospholipids, carbohydrates, and proteins, is the cell's protective boundary, facilitating selective substance exchange.
Phospholipids arrange themselves into a bilayer, with hydrophilic heads oriented outward and hydrophobic tails facing inward.
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Related Experiment Video

Updated: Aug 31, 2025

Assembly of Cell Mimicking Supported and Suspended Lipid Bilayer Models for the Study of Molecular Interactions
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Assembly of Cell Mimicking Supported and Suspended Lipid Bilayer Models for the Study of Molecular Interactions

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Determining small-molecule permeation through lipid membranes.

Jacopo Frallicciardi1, Matteo Gabba1, Bert Poolman2

  • 1Department of Biochemistry, University of Groningen, Groningen, the Netherlands.

Nature Protocols
|August 24, 2022
PubMed
Summary

This study presents a stopped-flow fluorimetry method to measure membrane permeability of water and solutes in lipid vesicles and yeast cells. The protocol provides a comprehensive toolbox for determining permeability coefficients efficiently.

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Author Spotlight: Advancing Cell Membrane Biophysics - Exploring Interactions and Challenges Through Experimental and Computational Approaches
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Area of Science:

  • Biophysics
  • Cell Biology
  • Biotechnology

Background:

  • Passive cell membrane permeability is crucial for understanding drug delivery, metabolism, and toxin effects.
  • Quantifying membrane permeability is essential in biology, biomedical research, and biotechnology.
  • Existing methods include radio-isotope distribution, spectroscopy, and molecular dynamics simulations.

Purpose of the Study:

  • To describe a protocol for estimating osmotic permeability of water and solutes across biological membranes using stopped-flow fluorimetry.
  • To provide a method for probing weak acid and base permeability in vesicles and cells.
  • To offer a comprehensive toolbox for determining membrane permeability coefficients.

Main Methods:

  • Stopped-flow fluorimetry measurements on lipid vesicles and living yeast cells.
  • Encapsulation of fluorescent dye (calcein) to monitor volume changes via fluorophore (de)quenching.
  • Utilizing pH-sensitive probes for weak acid and base permeability assessment.
  • Preparation of synthetic vesicles and dynamic light scattering for size distribution analysis.
  • Analysis of kinetic fluorescence data using MATLAB scripts.

Main Results:

  • The protocol enables the estimation of osmotic permeability coefficients for water and various solutes.
  • Weak acid and base permeability can be effectively probed in both model systems and living cells.
  • The method is adaptable to different lipid compositions and vesicle sizes.
  • Data analysis is facilitated by user-friendly MATLAB scripts.

Conclusions:

  • The described stopped-flow fluorimetry protocol offers a robust and versatile method for determining membrane permeability coefficients.
  • This approach provides valuable insights into the passive transport of molecules across biological membranes.
  • The protocol is suitable for a wide range of experimental systems and can be implemented with readily available equipment.