Chemical Permeabilization
Chemical Permeabilization - Related Videos
05:47
Oxygen Flux Measurements to Evaluate Mitochondrial Respiration in Living and Permeabilized Cells
0 Views •
2026
Source: Elettra Leo1, Luiza H.D. Cardoso1, Sabine Schmitt1, Erich Gnaiger11Oroboros InstrumentsThis video explains how oxygen flux is used to assess mitochondrial respiration in living and permeabilized mammalian cells. It demonstrates the sequential use of digitonin, metabolic substrates, ADP, an uncoupler, and an inhibitor to characterize different respiratory states. The resulting oxygen flux profile reveals mitochondrial function and energy-coupling efficiency under defined conditions.
08:21
Permeabilization of Adhered Cells Using an Inert Gas Jet
0 Views •
Cited by 3 •
2013
This protocol describes a method for the temporary permeabilization of adherent cells using an inert gas jet. This technique facilitates the transfer of genetic material and biomolecules into adherent mammalian cells by the utilization of mechanical forces to disrupt the plasma membrane.
05:30
Live-cell Imaging of Lysosomal Membrane Permeabilization During Necroptosis
0 Views •
2025
This protocol describes a live-cell imaging method for monitoring lysosomal membrane permeabilization during necroptosis, utilizing fluorescent probes that detect changes in lysosomal membrane integrity and acidification.
10:34
The Fabrication and Operation of a Continuous Flow, Micro-Electroporation System with Permeabilization Detection
0 Views •
Cited by 4 •
2022
This protocol describes the microfabrication techniques required to build a lab-on-a-chip, microfluidic electroporation device. The experimental setup performs controlled, single-cell-level transfections in a continuous flow and can be extended to higher throughputs with population-based control. An analysis is provided showcasing the ability to electrically monitor the degree of cell membrane permeabilization in real-time.
06:17
Measuring Mitochondrial Substrate Flux in Recombinant Perfringolysin O-Permeabilized Cells
0 Views •
Cited by 1 •
2021
In this work, we describe a modified protocol to test mitochondrial respiratory substrate flux using recombinant perfringolysin O in combination with microplate-based respirometry. With this protocol, we show how metformin affects mitochondrial respiration of two different tumor cell lines.
FAQs
Surfactants and organic solvents disturb the lipid bilayer, changing its permeability and creating transient pathways through which normally restricted molecules can pass. The intended effect is controlled disruption rather than permanent membrane failure. This mechanism enables intracellular access for proteins, nucleic acids, drugs, and fluorescent probes while preserving the possibility of subsequent cellular recovery.
Chemical concentration, exposure time, and recovery conditions jointly determine the balance between access and membrane damage. Increasing chemical exposure may improve molecular entry, but excessive disruption can reduce cell viability and alter cellular function. Bioengineering protocols therefore adjust these variables to obtain sufficient permeability without causing irreversible injury or compromising downstream analysis.
Transient disruption provides a temporary opportunity for external molecules to cross the membrane without requiring sustained loss of membrane integrity. That timing is important when delivering proteins, nucleic acids, drugs, or fluorescent probes into cells. Limiting the disruption helps preserve the biological system for later recovery, functional assessment, fixation, staining, or structural analysis.
Controlled permeabilization is limited in magnitude and duration, producing transient pathways that support molecular transport. Irreversible damage represents excessive membrane disruption and may reduce cell viability or change cellular function. This distinction makes exposure conditions central to experimental design: the objective is adequate molecular access while avoiding damage that could distort biological responses or analytical results.
A basic workflow involves exposing cells to a selected chemical under controlled conditions, allowing the desired molecular movement or intracellular access, and then applying appropriate recovery conditions. Concentration and exposure time require optimization for the cell system and intended use. Researchers can subsequently assess delivery, fixation, staining, or internal-structure analysis according to the experiment.
The main chemical choices described for this method are surfactants and organic solvents, selected to disturb the lipid bilayer in a controlled manner. Experimental planning must also specify the target molecule, chemical concentration, exposure time, and recovery conditions. These factors determine whether the treatment supports delivery or analysis while limiting effects on viability and function.
In bioengineering, this approach is useful when researchers need access to intracellular targets or internal structures. Applications include delivering proteins, nucleic acids, drugs, and fluorescent probes, as well as supporting cell fixation, staining, and analysis. Its value comes from combining temporary membrane access with conditions that aim to limit irreversible damage and preserve interpretable experimental outcomes.