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Detection, warping, and matching of the protein spots were done using the "combined warp and match" algorithm, which uses a nonparametric pattern recognition clustering technique to align different gel images.
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Identification of diffraction-limited fluorescence spots was done using SpotFinder, an accessory in MicrobeTracker, unless mentioned otherwise.
TLC was carrying out on silica gel-G, and spotting was done using UV light.
TLC analyses were done on glass plates coated with silica gel GF-254 and spotting was done using iodine/UV lamp.
The co-detection of the three CyDye-labeled forms for each spot was done using the Differential In-gel Analysis (DIA) module.
The blood spot analyses were done using the IMMUCHEM™ NEONATAL TSH-MW ELISA (bulk kit −20 plates) and RIA kits (500 tubes kit) provided by MP Biochemicals, USA.
Further imaging was done using a Zeiss LSM 510 Meta Confocal at 63× with 1-μm slices, and measurements of the number of GFP spots and distance were done using the LSM image browser software (release 4.0).
Optimization of the friction stir spot welding process parameters were done using response surface methodology, by constructing surface and contour plots.
Spot checking and intensity determination were done using ImaGene™ (BioDiscovery Inc., CA, USA).
Spot detection was done using ImageJ.
Spot recognition was done using Genepix Pro 7.0 (Molecular Devices).
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