Journal: Virulence

Article Title: Study of the host specificity of PB1-F2-associated virulence

doi: 10.1080/21505594.2021.1933848

Figure Lengend Snippet: Infection of embryonated eggs with WT and ΔF2 Nluc H7N1 viruses. Fertilized chicken eggs were infected at day 11 of embryonic development with 50 PFU of WT or ΔF2 Nluc H7N1 viruses. Eggs were processed for IVIS imaging at 24 and 48 h pi. Bioluminescence was acquired 5 min after injection of furimazine in the allantoic cavity (A) Embryos picture (top panel) and overlay of the NLuc signal (bottom panel) at 48 h pi. The scale on the right indicates the average radiance (sum of the photons per second from each pixel inside the ROI/number of pixels) p/sec/cm2/sr, number of photons per second per square centimeter per steradian. (B) Quantification of the NLuc signals at 24 and 48 h pi. Data are represented as the total flux in each pixel (p/s) integrated over the whole surface of the embryos. Between 5 and 10 eggs were monitored for each condition. Bars represent the geometric mean of a representative experiment performed twice

Article Snippet: Mice were anaesthetized and luminescence was measured 1 min after intra-nasal injection of 50 μl of PBS containing D-luciferin (7 mg.kg −1 , Synchem) or 50 μl of PBS containing furimazine (1:50 dilution of the Promega NanoGlo Luciferase Assay Substrate).

Techniques: Infection, Imaging, Injection

Journal: Virulence

Article Title: Study of the host specificity of PB1-F2-associated virulence

doi: 10.1080/21505594.2021.1933848

Figure Lengend Snippet: Kinetics of viral replication and induced inflammation in NF-κB-luciferase mice infected with WT or ΔF2 NLuc H7N1 viruses. NF-kB luciferase mice were infected with 10 5 PFU of WT or ΔF2 Nluc H7N1 intranasally. (A) Bioluminescence was measured by intranasal inoculation of furimazine or D-luciferin and using an IVIS system. NLuc signals (viral replication) were only observed in the nose and chest (left panels). Firefly luciferase (NF-κB activity) was monitored in the whole body (right panels). The color scales indicate the average radiance (sum of the photons per second from each pixel inside the ROI/number of pixels) p/sec/cm2/sr, number of photons per second per square centimeter per steradian. (B) Time course of NLuc activity is represented in the upper panel, time course of firefly activity is shown in the lower panel. Bioluminescence activities were quantified using Living Image software. Data are represented as the total flux in each pixel (p/s) integrated over the whole surface of the respiratory tract (NLuc) or the mouse (firefly). Curves are the mean ± SEM values obtained from five animals and are representative of at least three independent experiments

Article Snippet: Mice were anaesthetized and luminescence was measured 1 min after intra-nasal injection of 50 μl of PBS containing D-luciferin (7 mg.kg −1 , Synchem) or 50 μl of PBS containing furimazine (1:50 dilution of the Promega NanoGlo Luciferase Assay Substrate).

Techniques: Luciferase, Infection, Activity Assay, Software

Journal: Scientific Reports

Article Title: Ultrarapid Measurement of Diagnostic Antibodies by Magnetic Capture of Immune Complexes

doi: 10.1038/s41598-017-03786-7

Figure Lengend Snippet: Ultrarapid diagnosis of Sjögren syndrome using serum and saliva. Serum autoantibodies against Ro60 ( A ) and Ro52 ( B ) were detected by LIPSTICKS using Renilla luciferase fusion proteins in healthy controls (blue) and subjects with Sjögren’s syndrome (red). ( C ) Autoantibodies were also detected directly in 10 μl of saliva (without dilution) by LIPSTICKS in healthy controls (blue) and subjects with Sjögren’s syndrome (red). These results with saliva matched the ELISA using the corresponding serum from these individuals. ( D ) Ro52-NanoLuc fusion protein was used to measure autoantibodies by LIPSTICKS in healthy controls (blue) and patients with Sjögren’s syndrome (red). Shown is the comparison of the detection of the luciferase–generated light with the standard tube luminometer vs. the hand-held luminometer. The correlation coefficient between the values obtained with the two devices was R = 0.73 (p < 0.0001).

Article Snippet: Total luciferase activity in 1 μl of each crude extract was measured by adding it directly to 100 μl of assay buffer and substrate mixture ( Renilla Luciferase Reagent Kit, Promega or in the case of the Ro52-NanoLuc, the Nanoglow substrate, Promega) in a clear 1.5 ml microfuge tube, vortexing and immediately measuring in a luminometer (Turner design 20/20, Promega) for 1 s.

Techniques: Biomarker Discovery, Luciferase, Enzyme-linked Immunosorbent Assay, Comparison, Generated