Quantifying Membrane Order by Laurdan Staining

We analyzed differences in membrane order among cells with the fluorescent dye Laurdan. After treating various cells as described in the “Mitochondrial morphology assays” section, we stained cells (~ 70% confluency) with 5 μM Laurdan (Cayman Chemical, cat. 19706) as previously described [31 (link)] and imaged cells using two-photon microscopy (excitation 800 nm, emission 400–460 nm and 470–530 nm) [32 (link)]. We calculated general fluorescence polarization (GP) of Laurdan using custom MATLAB code as described previously [31 (link)].

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Humphries B.A., Cutter A.C., Buschhaus J.M., Chen Y.C., Qyli T., Palagama D.S., Eckley S., Robison T.H., Bevoor A., Chiang B., Haley H.R., Sahoo S., Spinosa P.C., Neale D.B., Boppisetti J., Sahoo D., Ghosh P., Lahann J., Ross B.D., Yoon E., Luker K.E, & Luker G.D. (2020). Enhanced mitochondrial fission suppresses signaling and metastasis in triple-negative breast cancer. Breast Cancer Research : BCR, 22, 60.

Publication 2020

Laurdan

Assays Cayman Cells Fluorescence polarization Fluorescent dye Laurdan Membrane Microscopy Mitochondrial

Corresponding Organization : University of Michigan–Ann Arbor

Other organizations : Michigan Center for Translational Pathology, University of California, San Diego

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Variable analysis

independent variables

Treatment of cells as described in the 'Mitochondrial morphology assays' section

dependent variables

Membrane order among cells, as measured by the fluorescent dye laurdan using two-photon microscopy and calculating the general fluorescence polarization (GP) of laurdan

control variables

Cell confluency (70%)
Laurdan concentration (5 μM)
Excitation wavelength (800 nm)
Emission wavelengths (400–460 nm and 470–530 nm)

controls

No positive or negative controls were explicitly mentioned.

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