9-month-old C57 mice (Woodley) were subcutaneously administered into the right footpad 1 mg of NIR815-labeled bNPs, 1 mg of NIR815-labeled bNPs pre-coated with Streptavidin 680RD, or streptavidin alone in 40 μL PBS. All animal use protocols were approved by Duquesne University Institutional Animal Care & Use Committee (IACUC). After 28 days, the popliteal draining lymph node from the injection-side foot, and spleens were collected from each mouse and imaged using an Odyssey M Imager (LI-COR) at 700 nm and 800 nm for streptavidin and bNPs, respectively.
Odyssey m imager
Manufactured by LI COR
The Odyssey M Imager is a compact, high-performance fluorescence imaging system designed for a variety of life science applications. It features a sensitive CCD camera and delivers accurate, quantitative data across a wide dynamic range. The Odyssey M Imager supports a range of sample types and detection modes, providing a versatile solution for researchers.
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Lab products found in correlation
Bicinchoninic acid assay, by Thermo Fisher Scientific (2 mentions)
Ripa buffer, by Thermo Fisher Scientific (2 mentions)
Ripa buffer, by G Biosciences (1 mentions)
Intercept blocking buffer, by LI COR (1 mentions)
Sc 7298, by Abcam (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
Trans blot turbo transfer system, by Bio-Rad (1 mentions)
Nupage 3 8 tris acetate gel, by Thermo Fisher Scientific (1 mentions)
Nitrocellulose membrane, by Merck Group (1 mentions)
Nupage transfer buffer, by Thermo Fisher Scientific (1 mentions)
Nupage lds sample loading buffer, by Thermo Fisher Scientific (1 mentions)
Nupage 4 12 bis tris gel, by Thermo Fisher Scientific (1 mentions)
Ecl detection reagent, by Bio-Rad (1 mentions)
Nupage mops sds running buffer, by Thermo Fisher Scientific (1 mentions)
Dithiothreitol (dtt), by Merck Group (1 mentions)
4 protocols using odyssey m imager
1
Biodistribution of Nanoparticles in Mice
2
RSF1 Expression Analysis in Embryos
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After injection with RSF1 MO or mRNA, embryos were collected at gastrula stages and lysed in lysis buffer [50 mM Tris-HCL (pH 7.5), 150 mM NaCl, 1 mM EDTA, 10% glycerol, 0.5% Triton X-100] with 10 μl buffer per embryo at 4°C on ice. The lysate was then centrifuged at 16,000 g for 15 min and the supernatant was used for SDS-PAGE, transferred to PVDF membrane. Immunoblots were performed with anti-RSF1 (Abcam ab109002, 1:1000), anti-GFP [Santa Cruz (B-2) sc-9996, 1:600], and anti-β-actin [Cell Signaling Technology (8H110D10) # 3700, 1:2000] antibodies. LI-COR Odyssey M imager was used to detect the signals.
3
Western Blot Analysis of Protein Expression
4
Western Blot Analysis of Protein Extracts
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Cells were washed in PBS, lysed directly in RIPA buffer (ThermoFisher) and protein extracts were quantitated using the Bicinchoninic acid assay (ThermoFisher) against a BSA standard curve. Extracts were made up in 4× NuPAGE LDS sample loading buffer (Invitrogen) supplemented with 100 mM dithiothreitol (Sigma-Aldrich), and incubated at 95°C for 10 min. Lysates (50–100 μg) were resolved by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) on NuPAGE 3–8% Tris–acetate gels (Invitrogen) in NuPAGE Tris–acetate running buffer (Invitrogen), or NuPAGE 4–12% Bis–Tris gels (Invitrogen) in NuPAGE MOPS SDS running buffer (Invitrogen). Gels were wet-transferred in 1× NuPAGE Transfer Buffer (Invitrogen), 20% ethanol and 0.05% SDS to nitrocellulose membranes (Millipore). 5% BSA/Tris-buffered saline + 0.01% Tween-20 (TBST) or 5% milk/TBST was used for blocking and incubation steps. Membranes were probed overnight at 4°C with indicated antibodies. The membrane was washed thrice for 5 min with TBST and incubated with HRP- or fluorescent dye-conjugated secondary antibodies for 1 h at room temperature. After four 5 min washes with TBST, HRP signals were detected with ECL detection reagent (BioRad) and imaged on an Amersham Imager 600RGB and fluorescence was imaged directly on a LI-COR Odyssey M Imager. Antibodies used in this study are described in Supplementary Table 4 .
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