The applied photosensitizers (PSs) were mesoporphyrin IX dihydrochloride (MPCl) and mesoporphyrin IX-dimethyl ester (MPE) (Frontier Scientific, Inc., Logan, UT). Their stock solutions were prepared in dimethyl-formamide (DMF) (Sigma, St. Louis, MO) with a concentration of around 2 mM and kept in a dark place (under 5 °C degree) to avoid photodegradation. Liposomes were made from saturated lipids as dimyristoyl-, dipalmitoyl- and distearoyl-phosphatidylcholine (DMPC, DPPC, DSPC) and unsaturated lipid as dioleoyl-phosphatidylcholine (DOPC) (Sigma, St. Louis, MO). The applied fluorescent dye for fluorescence correlation spectroscopy (FCS) was 3,3′-Dioctadecyloxacarbocyanine Perchlorate (DiO) (Sigma, St. Louis, MO).
Dioleoylphosphatidylcholine
Manufactured by Merck Group
Dioleoylphosphatidylcholine is a phospholipid commonly used in laboratory research. It is a synthetic version of a naturally occurring phospholipid found in cell membranes. Dioleoylphosphatidylcholine serves as a structural component in model lipid bilayers and liposomes, which are used to study membrane-related processes.
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Lab products found in correlation
4 protocols using dioleoylphosphatidylcholine
1
Photosensitizer Liposome Formulation for FCS
2
Fluorescent Lipid Membrane Protocol
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Dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC) and cholesterol were purchased from Sigma-Aldrich. Bovine brain sphingomyelin (SM) was from Avanti Polar Lipids (Alabaster, USA). Laurdan was purchased from Life Technologies. Dye PA was synthesized as described elsewhere35 (link).
3
Characterization of CGA-N9 Membrane Disruption on Candida
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The effect of CGA-N9 on the membrane integrity of C. tropicalis was characterized according to the leakage rate of calcein from liposomes. Calcein-loaded liposomes were prepared according to a previously reported method [34 (link)] with minor modifications. Dioleoyl phosphatidylcholine and dioleoyl phosphoethanolamine (Sigma–Aldrich, Shanghai, China) (1 : 1.27, w/w) were employed to mimic the logarithmic-phase Candida lipid bilayer [35 (link)]. A suspension of calcein-loaded liposomes was incubated with 3.9 µg/ml CGA-N9 at 28°C to evaluate pore formation and membrane integrity by measuring dye leakage every 1 h using a fluorescence spectrophotometer (Cary Eclipse, CA, U.S.A.) at λex/λem = 492/517 nm. In this assay, 20 mmol/l PBS (pH 7.0) was selected as a negative control, and 10 mmol/l H2O2 was used as a positive control. All samples were examined in triplicate. After 10 h in the presence of 0.1% Triton X-100 in Tris buffer, 100% of the dye was released from liposomes.
4
Influenza hemagglutinin binding assay
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DPPE was purchased from Wako Pure Chemical Industries, Ltd. (Japan). Dioleoylphosphatidylcholine (DOPC) and 1-palmitoyl-2-oleoyl-sn-glycelo-3-phosphocholine (POPC) were purchased from Sigma-Aldrich Co., LLC (St. Louis, MO, USA). Ganglioside Neu5Acα2–3Galβ1–4Glcβ1–1′Cer (GM3) from the bovine brain was purchased from Hytest Ltd. (Finland). Influenza hemagglutinins (HAs) derived from A/New Caledonia/20/99 (H1N1) and A/New York/55/2004 (H3N2) viruses were provided by Yujiro Suzuki (The Kitasato Institute, Japan; Matsubara et al., 2010 (link)). The human IFV strain A/Puerto Rico/8/34 (H1N1) was provided by Dr. Kyosuke Nagata (University of Tsukuba, Japan; Matsubara et al., 2009 (link)).
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