Visualizing Tension and Growth in Model Membranes Using Optical Dyes.

Biophysical journal (2018-09-17)
Margrethe A Boyd, Neha P Kamat
ABSTRACT

Cells dynamically regulate their membrane surface area during a variety of processes critical to their survival. Recent studies with model membranes have pointed to a general mechanism for surface area regulation under tension in which cell membranes unfold or take up lipid to accommodate membrane strain. Yet we lack robust methods to simultaneously measure membrane tension and surface area changes in real time. Using lipid vesicles that contain two dyes isolated to spatially distinct parts of the membrane, we introduce, to our knowledge, a new method to monitor the processes of membrane stretching and lipid uptake in model membranes. Laurdan, located within the bilayer membrane, and Förster resonance energy transfer dyes, localized to the membrane exterior, act in concert to report changes in membrane tension and lipid uptake during osmotic stress. We use these dyes to show that membranes under tension take up lipid more quickly and in greater amounts compared to their nontensed counterparts. Finally, we show that this technique is compatible with microscopy, enabling real-time analysis of membrane dynamics on a single vesicle level. Ultimately, the combinatorial use of these probes offers a more complete picture of changing membrane morphology. Our optical method allows us to remotely track changes in membrane tension and surface area with model membranes, offering new opportunities to track morphological changes in artificial and biological membranes and providing new opportunities in fields ranging from mechanobiology to drug delivery.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Calcein, Used for the fluorometric determination of calcium and EDTA titration of calcium in the presence of magnesium.
Sigma-Aldrich
6-Dodecanoyl-N,N-dimethyl-2-naphthylamine, suitable for fluorescence, ≥97.0% (HPLC)