Abstract
The mechanism of surfactant-induced cell lysis has been studied with quantitative coherent anti-Stokes Raman scattering (CARS) microspectroscopy. The dynamics of surfactant molecules as well as intracellular biomolecules in living Chinese Hamster Lung (CHL) cells has been examined for a low surfactant concentration (0.01 w%). By using an isotope labeled surfactant having CD bonds, surfactant uptake dynamics in living cells has been traced in detail. The simultaneous CARS imaging of the cell itself and the internalized surfactant has shown that the surfactant molecules is first accumulated inside a CHL cell followed by a sudden leak of cytosolic components such as proteins to the outside of the cell. This finding indicates that surfactant uptake occurs prior to the cell lysis, contrary to what has been believed: surface adsorption of surfactant molecules has been thought to occur first with subsequent disruption of cell membranes. Quantitative CARS microspectroscopy enables us to determine the molecular concentration of the surfactant molecules accumulated in a cell. We have also investigated the effect of a drug, nocodazole, on the surfactant uptake dynamics. As a result of the inhibition of tubulin polymerization by nocodazole, the surfactant uptake rate is significantly lowered. This fact suggests that intracellular membrane trafficking contributes to the surfactant uptake mechanism.
Highlights
Interactions of surfactants with living cells are of considerable interest with regard to their biological functions including cellular toxicity [1]
Previous studies have shown that microorganisms solubilization by surfactants occurs with cell lysis, in which the cell membrane is degraded by surfactants with eventual breakdown of the whole cell [5,6,7,8]
We use a recentlyemerging new tool, coherent anti-Stokes Raman scattering (CARS) microspectroscopy [9,10,11,12,13], which is powerful for studying lipid molecules in living cells
Summary
Interactions of surfactants with living cells are of considerable interest with regard to their biological functions including cellular toxicity [1]. Understanding their toxicological mode of action is highly important in order to assess and control their safety on human exposure [2,3,4]. The dynamical process of surfactant action in single living cells is still unexplored because of the lack of the mean to visualize surfactant molecules in vivo and in situ. We use an isotope labeled surfactant (d25-sodium dodecyl sulfate (SDS)) and visualize the dynamics of surfactant molecules in the cell lysis process. Deuterium substitution enables us to selectively trace the SDS molecules among a number of unlabeled biomolecules [10,14,15,16]. d25-SDS gives CD stretch bands in the 2000– 2200 cm spectral region, which is a ‘‘window’’ of Raman spectra of unlabeled biomolecules, facilitating its selective detection
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