RNA extractions and transcriptomics analyses were performed at HistoGeneX (Antwerp, Belgium). Tumor tissue was macro-dissected from 5 μm sections of FFPE tissues and total RNA was extracted using the High Pure RNA FFPE Micro Kit (Roche, Mannheim, Germany), following the manufacturer’s protocol. RNA was quantified using the Ribogreen kit (high range; Molecular Probes Inc.; Eugene, OR) on a Fluoroskan Ascent instrument (ThermoFisher Scientific; Waltham, MA). Seven standard dilution points ranging from 20 ng/mL to 1000 ng/mL were used to quantify samples across a wide range of concentrations. Sample purity was evaluated by measuring 260/280 nm and 260/230 nm ratios using a Nanodrop™ spectrophotometer (ThermoFisher). All samples were titrated to 14 ng/μL, and 100 ng total sample RNA was analyzed on the NanoString nCounter® system (NanoString; Seattle, WA), which consist of the nCounter® Prep Station and nCounter® Digital Analyzer (NanoString), using a commercially available pre-defined probe set (nCounter PanCancer Immune Profiling Panel, NanoString). Data normalization was done using the nSolver® Analysis Software (NanoString) [16 (link)]. Data were analyzed via nSolver software and R v3.3.1 (R Foundation for Statistical Computing) [17 ].
Fluoroskan ascent instrument
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Fluoroskan Ascent is a microplate fluorometer and luminometer instrument designed for high-throughput fluorescence and luminescence measurements. It features a monochromator-based optical system and a temperature-controlled measurement chamber. The instrument is capable of performing various fluorescence and luminescence detection assays in microplates.
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Lab products found in correlation
2 protocols using fluoroskan ascent instrument
1
FFPE RNA Extraction and Immune Profiling
2
Fecal Hemoglobin Quantification Protocol
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Haemoglobin in faeces was determined from 20–30 mg of a well‐mixed faecal sample as described previously.18 Following several extraction and purification steps, 200 μL of aqueous phase from the final step was transferred into a black 96‐microwell plate and measured (excitation wavelength 399 nm; emission wavelength 614 nm) with a Fluoroskan Ascent instrument (Thermo Labsystems, Philadelphia, PA, USA). Porcine haemoglobin (H4131, Sigma‐Aldrich, St. Louis, MO, USA) in Drabkin's reagent (D5941, Sigma‐Aldrich) was used as a reference standard.
Blood haemoglobin was analysed at baseline and at the end of each study phase at a local hospital laboratory (Tykslab, Turku, Finland) with standard clinical laboratory methods after 10 hours fasting before sampling.
Blood haemoglobin was analysed at baseline and at the end of each study phase at a local hospital laboratory (Tykslab, Turku, Finland) with standard clinical laboratory methods after 10 hours fasting before sampling.
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