Mean fluorescence anisotropy variations over time for eGFP and eGFP-tandem.

<p>Mean fluorescence anisotropy decays for eGFP (<i>solid square</i>, n = 19 cells) and eGFP-tandem (<i>open round</i>, n = 21 cells) expressed in HEK293 cells.</p

Fluorescence intensity decays obtained for parallel and perpendicular polarizations using objective 10× (NA = 0.3).

<p>Polarizations were normalized with the excitation polarization. Values obtained at each time gate for (<i>open squares</i>) and (<i>stars</i>) are represented. <i>(A)</i> non-viscous fluorescein solution. <i>(B)</i> fluorescein solution with a high viscosity (69–70% glycerol).</p

Anisotropy measurements for APP-eGFP and L17C APP-eGFP expressed in HEK293 cells in TIRF excitation.

<p><i>(A)</i> Mean fluorescence anisotropy variation over time for APP-eGFP (<i>solid square</i>, n = 8 cells) and L17C APP-eGFP (<i>open round</i>, n = 9 cells). <i>(B)</i> Representative examples of mean fluorescence intensities with polarization parallel (<i>central panel</i>) or perpendicular <i>(left panel</i>) to the excitation one and mean photon-weighted fluorescence anisotropy map (<i>right panel</i>) for APP-eGFP (<i>upper panel</i>) or L17C APP-eGFP (<i>lower panel</i>). These images were averaged on all time gates. The scale bar represents 10 µm.</p

Schematic of the tr-FAIM coupled to a TIRFLIM set-up.

<p><i>(A)</i> Schematic of the set-up. SC: supercontinuum fiber laser. M1 & M2: cold mirrors to filter the infrared part of the laser. OD: optical density to control the laser power, which is typically of 500 µW (measured in the back focal plane of the objective). L1 & L2: telescope to enlarge the beam to the field of view. F: filter. L3: lens to focus the beam in the back focal plane of the objective 60<b>×</b> (NA = 1.49). TM: mirror mounted on a translation stage to switch from an epifluorescence excitation to a TIRF excitation with different penetration depths. HRI: high rate imager. CCD: charge-coupled device camera. <i>(B)</i> tr-FAIM acquisition. Temporal gates are opened at different times after the laser pulse, resulting in a series of time-gated images for each polarization.</p

Fluorescence anisotropy decays of fluorescein acquired with objectives of different NA.

<p>Fluorescein solution of high viscosity (69–70% glycerol) was probed using objective 10<b>×</b>, NA = 0.3 (<i>solid square</i>), objective 40<b>×</b>, NA = 1 (<i>open round</i>) and objective 60<b>×</b>, NA = 1.49 (<i>solid triangle</i>).</p

Baseline characteristics of case and control subjects [number (percentage) or mean ± standard deviation].

<p>Baseline characteristics of case and control subjects [number (percentage) or mean ± standard deviation].</p

Association between menstrual and reproductive factors and breast cancer risk.

<p>Association between menstrual and reproductive factors and breast cancer risk.</p