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Freeze–thaw immobilization of liposomes in chromatographic gel beads: evaluation by confocal microscopy and effects of freezing rate

✍ Scribed by Andreas Lundqvist; Göran Ocklind; Lars Haneskog; Per Lundahl


Publisher
John Wiley and Sons
Year
1998
Tongue
English
Weight
151 KB
Volume
11
Category
Article
ISSN
0952-3499

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✦ Synopsis


Biological membranes immobilized in chromatographic gel beads constitute a multifunctional affinity matrix. Membrane protein-solute interactions and drug partitioning into the lipid bilayers can conveniently be studied. By the use of confocal laser-scanning microscopy (CLSM) the distribution of immobilized model membranes in the beads has been visualized for the first time. Freeze-thaw-immobilized liposomes in Superdex 200 gel beads were situated in a thick shell surrounding a liposome-free core. The amount of phospholipids immobilized by freeze-thawing was dependent on the temperature in the cooling bath and the type of test tube used. A bath temperature of À25 °C gave higher immobilization yield than freezing at À75 or À8 °C did. Freeze-thawing in the presence of liposomes did not affect the gel bead shape or the refractive index homogeneity of the agarose network of the beads, as shown by confocal microscopy.