Fig. 2
From: Line excitation array detection fluorescence microscopy at 0.8 million frames per second
Resolution and detection limit characterization of the LEAD microscope. Fluorescent beads imaged within the device resulted in PSFs ranging between (a) 3.4–4.7 μm (FWHM) in the z-direction and (b) 3.0–4.1 μm (FWHM) in the x-direction across the FOV. No beads were detected along some edges of the device in (b) due to the parabolic flow velocity profile of beads in the device. c The detection limit is determined by measuring the signal-to-noise ratio (SNR) of different concentrations of fluorescein flowing in the device. An SNR model for PMTs gives a collection efficiency of the system of 2.9 ± 1.2% and a detection limit (SNR = 1) of 22.3 ± 9.0 nM, corresponding to 420 ± 170 molecules of fluorescein imaged by each PMT element. Uncertainty arises from the varying signal level across experimental repeats. d Signal-to-background ratios (SBR) of polyQ40 strain C. elegans moving at 1 m s−1 imaged with our platform. SBR exceeds 200 for the center PMT elements and drops off for surrounding elements due to their decreased responsivity. Uncertainty arises from the varying fluorescence for different samples. For all panels, data are mean and error bars are s.d.
