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Image Search Results
Journal: bioRxiv
Article Title: Correlative cryo-structured illumination fluorescence microscopy and soft X-ray tomography elucidates reovirus intracellular release pathway
doi: 10.1101/2020.01.13.904623
Figure Lengend Snippet: a , Widefield and b , SIM lateral point spread of a 175nm diameter, 505/515nm wavelength microsphere (PS-SpeckTM ThermoFischer) collected on the cryo-SIM at 71K using 488nm light for excitation and a 520/25nm filter (Semrock) for emission. c , Plot of lateral point spread (single line profile) of a , and b , and their respective gaussian fits showing clear resolution enhancement. d , SPEKcheck display of the cryo-SIM for a representative fluorophore (Alexa488) excited in the system with an efficiency of 68.7% and a resulting collection of 69.9% of the total emitted light. Laser light is shown as a bright blue spike, excitation and emission are shown in shades of light green and the total transmitted light is shown in bright green. The system dichroic mirrors and filters are also represented (system dichroics denoted as ZT, detector splitter dichroic as T560 and the emission filter as EM). Analysis of representative cryo-SIM data from a single mammalian U2O9 cell containing structures tagged with Alexa488 fluorophore: e, Z axis projection of the raw SIM data processed to produce a pseudo-widefield image stack; f, Z axis projection of the same SIM data fully reconstructed showing the resulting resolution enhancement. g , Intensity analysis of plots of h-k (flattened and sampled radially) showing the relative increase in information content after reconstruction. h and i , Reciprocal space resolution plots of the raw pseudo-widefield data in the cell shown in e (Alexa488 fluorescence and mCherry fluorescence (associated with a tagged reporter molecule) respectively). j and k , Same data as h and I respectively after SIM reconstruction. Concentric dashed circles for h - k denote resolution boundaries of 600nm (small circle) and 200nm ((large circle). All image analysis was done using Fiji and the SIMcheck Fiji plugin.
Article Snippet: To better understand reovirus escape from endocytic vesicles, we have used the
Techniques: Fluorescence