nis elements general analysis 3 pipeline  (Nikon)

Journal: Nature Neuroscience

Article Title: Phosphorylated tau exhibits antimicrobial activity capable of neutralizing herpes simplex virus 1 infectivity in human neurons

doi: 10.1038/s41593-025-02157-0

Figure Lengend Snippet: 2D ReNcell VM cultures were preincubated with dilutions of synthetic 2N3R tau, 2N4R tau or 2N4R GSK-3β p-tau followed by HSV-1 infection to assay the antimicrobial protective properties of tau. After HSV-1 infection, whole wells were imaged by confocal microscopy and analyzed for red fluorescence. a , Images comparing HSV-1 plaque formation in the absence or presence of 2N4R GSK-3β p-tau (1.25 µg ml −1 ). b − f , Whole-well images from the 2N4R GSK-3β p-tau (scale of 2.5 µg ml −1 to 0.625 µg ml −1 ) pretreated conditions were analyzed using Nikon Elements GA3 to compare the number of HSV-1 plaques ( F 3, 54 = 7.069, ** P = 0.0015) ( b ), the number of HSV-1 single-cell infections ( F 3, 54 = 2.754, * P = 0.0291) ( c ), the ratio of plaques to single-cell infections ( F 3, 54 = 6.428, * P = 0.0169) ( d ), the size of individual HSV-1 plaques by area (* P = 0.0260) ( e ), and the average size of HSV-1 plaques by area distributed into quadrants ( F 3, 1,376 = 1.859, ** P = 0.0022) ( f ). In addition, whole well images from the 2N3R and 2N4R tau conditions (1.25 µg ml −1 ) were analyzed using Nikon Elements General Analysis 3 (GA3) to compare the number of HSV-1 plaques ( g ) and the number of HSV-1 single-cell infections ( h ). Box plots are representative of ±s.e.m. ( n = 11) depicting median and interquartile range, with whiskers denoting variability according to Tukey’s method. Statistical mean comparisons were calculated by one-way ANOVA using Dunnett’s multiple comparisons test ( b − d ), one-way ANOVA using Tukey’s multiple comparisons test ( g , h ), two-tailed Kolmogorov−Smirnov test ( e ) for plaque size distribution and two-way ANOVA using Sidak’s multiple comparisons test ( f ).

Article Snippet: Nikon NIS Elements General Analysis 3 workflows or ‘recipes’ for image analysis are available from the corresponding authors upon reasonable request.

Techniques: Infection, Confocal Microscopy, Fluorescence, Single Cell, Two Tailed Test

Journal: Nature Neuroscience

Article Title: Phosphorylated tau exhibits antimicrobial activity capable of neutralizing herpes simplex virus 1 infectivity in human neurons

doi: 10.1038/s41593-025-02157-0

Figure Lengend Snippet: ReNcell VM cultures were infected with HSV1 for 24 hours to characterize total tau’s distribution changes from infection and in proximity to viral infection. ( a ) 2D ReNcell VM cultures in microfluidic devices were infected in the left chamber with HSV1 for 48 hours, immunoprobed with anti-total tau labeled with a fluorescent secondary antibody, and analyzed for total tau (405), neurons (GFP), and HSV1 (RFP) fluorescence by confocal microscopy. Fluorescence signals were compared by General Analysis 3 (GA3). ( b ) GA3 of HSV1-positive neurons compared intracellular p-tau fluorescence between infected neurons, uninfected neurons proximal to infected neurons, and uninfected neurons not proximal to infected neurons (F (2, 3321) =166.3, **** P <0.0001). ( c ) Individual neuronal total tau fluorescent intensity was compared to total proximal HSV1-RFP fluorescent intensity (R²=1.350e-006). ( d ) Fluorescent image captures from 384 wells over 4 experiments were compared for total tau fluorescence in cell bodies (*** P =0.0003) between uninfected and infected wells. Statistical mean comparisons were calculated by one-way ANOVA ( b ), simple linear regression ( c ), and two-tailed paired t-test ( d ). Images are representative of experiments repeated in triplicate.

Article Snippet: Nikon NIS Elements General Analysis 3 workflows or ‘recipes’ for image analysis are available from the corresponding authors upon reasonable request.

Techniques: Infection, Labeling, Fluorescence, Confocal Microscopy, Two Tailed Test