Fig. 3: The sensitivity of detectors varies with wavelength and determines the digital resolution and image quality.
From: Best practices and tools for reporting reproducible fluorescence microscopy methods
a, Fluorescence images of a pattern consisting of lines with incrementally increasing spacing on the ArgoLight-SIM calibration slide acquired with the same objective and a camera with either a 6.5-µm (left) or 16-µm (right) photodiode size. Lower row, line intensity profile plots. Scale bar, 2.5 µm. b, Fluorescence images (using a Fire LUT) of BPAE cells stained with Alexa Fluor 488 phalloidin acquired under identical imaging conditions with a single-point scanning confocal microscope using either a GaAsP (left) or multialkali PMT (MA; center) detector. Images are displayed with the same intensity scale. Right, fluorescence intensity quantification; bars represent the population mean. Scale bar, 10 µm. c, Fluorescence images (using a Fire LUT) of cultured cells prepared by Leica and imaged under identical conditions with a Leica Stellaris 8 Power HyD S (left column), Power HyD X (center-left column) or Power HyD R (center-right column) show a difference in intensity according to wavelength (right). Green, HyD S; purple, HyD X; yellow, HyD R. Scale bars, 10 µm. d, Fluorescence images of a pattern consisting of a repeating series of lines with progressively decreasing intensity (pattern C) on the ArgoLight-SIM calibration slide were acquired with an Andor Zyla sCMOS 4.2 plus camera under low-light conditions (left panels) or high-light conditions (right panels). Images were acquired under identical conditions in each case, with the exception of varying the amplifier gain amplification. Top left and top right, low gain; bottom left and bottom right, high gain. Bottom panels, plots of fluorescence intensities along a vertical line scan through the center of the pattern (purple, low gain amplification; green, high gain amplification). Scale bar, 5 µm.
