p200 pipet  (Bio-Techne corporation)

Journal: Science Advances

Article Title: Single-dose mucosal replicon-particle vaccine protects against lethal Nipah virus infection up to 3 days after vaccination

doi: 10.1126/sciadv.adh4057

Figure Lengend Snippet: Kaplan-Meier survival curves of animals inoculated with NiV∆F: ( A ) Two- to 3-day-old suckling mice were intracerebrally (IC) inoculated with either 10 5 or 10 3 TCID 50 of NiV∆F or one of seven dilutions of wild-type NiV ranging from 10 6 to 10 0 TCID 50 . ( B ) Five- to 7-week-old Syrian hamsters were IN inoculated with either 10 6 TCID 50 of NiV∆F or one of three dilutions of wild-type NiV ranging from 10 6 to 10 3 TCID 50 . Survival significance calculated by log-rank (Mantel-Cox test): **** P ≤ 0.0001; *** P ≤ 0.001; ** P ≤ 0.01; * P ≤ 0.05. ( C ) Differential mRNA expression of select innate immune response genes in lungs of animals ( D to G ) was determined by RT-qPCR. (D) and (E) Lung and brain tissues and (F) and (G) mucosal swab samples (oral and rectal) were taken at euthanasia from hamsters serially euthanized 1, 3, or 7 days after inoculation with 10 6 TCID 50 of NiV∆F (green circles) or 1, 3, or 5 to 7 days after inoculation with wild-type NiV (red squares) to determine both vRNA levels (RT-qPCR) and quantify infectious virus titers (TCID 50 ) in sample types in which NiV∆F RNA was detected. For samples analyzed in (C) to (G), a subset of wild-type NiV-infected animals did not survive to predetermined 7 dpi time point and were sampled earlier when end point criteria were reached (5 to 6 dpi). Significance was calculated by multiple t test: * P ≤ 0.05; ** P ≤ 0.01.

Article Snippet: For vaccination with NiV∆F, hamsters (male and female; 5- to 7-week-old HsdHan:AURA Syrian hamsters; Envigo, 8903F or 8903M) were inoculated IN with 10 6 TCID 50 (100 μl divided bilaterally between the nares using a P200 pipette (Rainin Pipet-Lite XLS LTS).

Techniques: Expressing, Quantitative RT-PCR, Virus, Infection

Journal: Science Advances

Article Title: Single-dose mucosal replicon-particle vaccine protects against lethal Nipah virus infection up to 3 days after vaccination

doi: 10.1126/sciadv.adh4057

Figure Lengend Snippet: Lung tissues from hamsters serially euthanized following inoculation with 10 6 TCID 50 of NiV∆F (1, 3, or 7 days), wild-type NiV [1, 3, or 5 to 7 days; subset of wild-type NiV-infected animals did not survive to predetermined 7 dpi time point and were sampled earlier when end point criteria were reached (5 to 6 dpi)], or DMEM only (mock-infected at 1, 3, or 7 days) were formalin fixed and evaluated via ( A ) hematoxylin and eosin staining to characterize tissue pathology and via ( B ) ISH to detect vRNA. Lungs of NiV∆F- and mock-inoculated animals showed no notable histopathologic changes at any time point, while lungs from NiV-inoculated animals displayed progressive inflammation, with epithelial syncytia (insets) compatible with paramyxoviral pneumonia. NiV∆F vRNA was detected by ISH 1 dpi and decreased by 7 days. In contrast, NiV vRNA was detected at relatively higher levels starting 1 day after inoculation and increased over time with severity of pneumonia.

Article Snippet: For vaccination with NiV∆F, hamsters (male and female; 5- to 7-week-old HsdHan:AURA Syrian hamsters; Envigo, 8903F or 8903M) were inoculated IN with 10 6 TCID 50 (100 μl divided bilaterally between the nares using a P200 pipette (Rainin Pipet-Lite XLS LTS).

Techniques: Infection, Staining

Journal: Science Advances

Article Title: Single-dose mucosal replicon-particle vaccine protects against lethal Nipah virus infection up to 3 days after vaccination

doi: 10.1126/sciadv.adh4057

Figure Lengend Snippet: Five- to 7-week-old Syrian hamsters were inoculated either IN (green circles) or SC (blue triangles) with 10 6 TCID 50 of NiV∆F and euthanized 1, 3, 7, 14, or 28 days postvaccination (dpv; n = 10 per group). ( A ) NiV∆F vRNA tissue levels at each time point were determined by RT-qPCR. ( B ) IgG antibody titers against NiV N and G proteins were determined by enzyme-linked immunosorbent assay (ELISA) at all time points after vaccination. ( C ) For IN-vaccinated hamsters, IgA titers against NiV F, N, and G were determined by ELISA at 28 days postvaccination. ( D ) Neutralizing antibody titers against NiV strain Malaysia were evaluated 1, 3, 7, 14, and 28 days postvaccination. ( E ) Antibody-dependent complement deposition (ADCD) assay showing complement fixing activity of antibodies and ( F ) antibody-dependent cellular phagocytosis (ADCP) assay depicting phagocytic activity of antibodies were performed on plasma collected 7, 14, and 28 days postvaccination. In all panels, individual values are shown, with bars representing mean and SD. Significance calculated by multiple t test: * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001. ND, not detected; BLD, below limit of detection; MFI, mean fluorescence intensity.

Article Snippet: For vaccination with NiV∆F, hamsters (male and female; 5- to 7-week-old HsdHan:AURA Syrian hamsters; Envigo, 8903F or 8903M) were inoculated IN with 10 6 TCID 50 (100 μl divided bilaterally between the nares using a P200 pipette (Rainin Pipet-Lite XLS LTS).

Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Activity Assay, Clinical Proteomics, Fluorescence

Journal: Science Advances

Article Title: Single-dose mucosal replicon-particle vaccine protects against lethal Nipah virus infection up to 3 days after vaccination

doi: 10.1126/sciadv.adh4057

Figure Lengend Snippet: Groups of 10 hamsters were vaccinated IN with 100 μl containing 10 6 TCID 50 of NiV∆F: once 28 (black), 14 (orange), 7 (purple), or 3 (green) days before challenge; or twice (blue) both 3 and 1 days before challenge. Mock-vaccinated animals (red) were given an equivalent IN volume of DMEM 28 days before challenge. All hamsters were challenged IN with 10 6 TCID 50 NiV strain Malaysia. ( A ) Percent weight change from baseline (taken −1 dpi); mean daily body temperatures; and individual daily clinical scores (from 0 to 10), with severity depicted by increased intensity of red. Animals scoring ≥ 10 were humanely euthanized; any animal that succumbed to disease before euthanasia was allocated a score of 10. Gray boxes indicate the end of monitoring/scoring due to euthanasia/death. Individual animals are represented as circles, with the solid line representing the daily mean. ( B ) Mean percent weight changes from baseline (taken −1 dpi; significance calculated by two-way analysis of variance (ANOVA): * P ≤ 0.05). ( C ) Kaplan-Meier curves showing survival of vaccinated and mock-vaccinated animals challenged with NiV strain Malaysia. Significance calculated by log-rank (Mantel-Cox test): * P ≤ 0.05; ns, not significant. ( D ) RT-qPCR detection of NiV vRNA in select tissues from hamsters vaccinated at indicated times and subsequently challenged. Individual animals from each group are represented. Error bars represent the means and SD. Significance calculated by t test: **** P ≤ 0.0001; *** P ≤ 0.001; ** P ≤ 0.01; * P ≤ 0.05.

Article Snippet: For vaccination with NiV∆F, hamsters (male and female; 5- to 7-week-old HsdHan:AURA Syrian hamsters; Envigo, 8903F or 8903M) were inoculated IN with 10 6 TCID 50 (100 μl divided bilaterally between the nares using a P200 pipette (Rainin Pipet-Lite XLS LTS).

Techniques: Quantitative RT-PCR

Journal: Science Advances

Article Title: Single-dose mucosal replicon-particle vaccine protects against lethal Nipah virus infection up to 3 days after vaccination

doi: 10.1126/sciadv.adh4057

Figure Lengend Snippet: Groups of 9 and 10 hamsters were vaccinated IN with 100 μl containing 10 6 TCID 50 of NiV∆F: once 28 (black), 14 (orange), 7 (purple), or 3 (green) days before challenge; or twice (blue) both 3 and 1 days before challenge. Mock-vaccinated animals (red) were given an equivalent IN volume of DMEM 28 days before challenge. All hamsters were challenged IP with 10 4 TCID 50 NiV strain Malaysia. ( A ) Percent weight change from baseline (taken −1 dpi); mean daily body temperature; and individual daily clinical scores (scored from 0 to 10), with severity depicted by increased intensity of red. Animals scoring ≥ 10 were humanely euthanized; any animal that succumbed to disease before euthanasia was allocated a score of 10. Gray boxes indicate the end of monitoring/scoring due to euthanasia or death. Individual animals are represented as circles, with the solid line representing the daily mean. ( B ) Mean percent weight changes from baseline (taken at −1 dpi; significance calculated by two-way ANOVA: * P ≤ 0.05). ( C ) Kaplan-Meier curves showing survival of vaccinated and mock-vaccinated animals challenged with NiV strain Malaysia. Significance calculated by log-rank (Mantel-Cox test): **** P ≤ 0.0001; ** P ≤ 0.01. ( D ) RT-qPCR detection of NiV vRNA in select tissues from hamsters vaccinated at indicated times and subsequently challenged. Individual animals from each group are represented. Error bars represent the means and SD. Significance calculated by t test: **** P ≤ 0.0001; *** P ≤ 0.001; ** P ≤ 0.01.

Article Snippet: For vaccination with NiV∆F, hamsters (male and female; 5- to 7-week-old HsdHan:AURA Syrian hamsters; Envigo, 8903F or 8903M) were inoculated IN with 10 6 TCID 50 (100 μl divided bilaterally between the nares using a P200 pipette (Rainin Pipet-Lite XLS LTS).

Techniques: Quantitative RT-PCR

Journal: STAR Protocols

Article Title: Fluorescence-activated cell sorting and phenotypic characterization of human fibro-adipogenic progenitors

doi: 10.1016/j.xpro.2022.102008

Figure Lengend Snippet:

Article Snippet: Pipetting tips, P200 , Sarstedt , Cat#70.760.502.

Techniques: Recombinant, Blocking Assay, Saline, Imaging, Software, Fluorescence, FACS, Flow Cytometry, Dissection, Transferring, Cell Culture, Sterility