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The patient's head was fixed firmly but comfortably to minimise movement during the PET-CT scan using the standard scanner headrest.
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This allowed us to create a custom acquisition sequence that first acquired a single Z-stack in 3D-SIM (prebleached image), then performed single or multi spot photobleaching (using the standard OMX galvo scanner TIRF/photo-kinetics module), then performed time lapse imaging in widefield mode (including the photobleached image), and then performed a second 3D-SIM Z-stack (5 min recovery image).
This allowed us to create a custom acquisition sequence that first acquired a single Z-stack in 3D-SIM, then performed single or multiple spot photobleaching (using the standard OMX galvo scanner TIRF/photo-kinetics module), and then performed time lapse imaging in 3D-SIM mode.
To facilitate image analysis, multiplanar reformations in the coronal plane and sagittal plane (section thickness, 3 mm) were calculated using the standard software provided with the scanner.
Hybridization was carried out for 17 hours at 65°C using a Microarray hybridization chamber (Agilent Technologies) in a rotating hybridization oven prior to washing per Agilent protocol and scanning with an Agilent Scanner (DNA Microarray Scanner, Agilent Technologies) using the standard parameters for a gene expression 8 × 15K oligoarray (5 μm and 20 bits).
The scans were obtained with an advanced high-field 1.5-T scanner (Sonata Magnetom, Siemens) using the standard protocol.
The relaxation time T1 was measured using the standard spin-echo sequence on a 3T MR scanner with a volume head coil as RF receiver operating at 37°C.
All scans were acquired on the same 1.5T GE Signa MR scanner (GE Medical Systems, Milwaukee, WI) using the standard circularly polarised quadrature birdcage coil.
Washing and staining were performed in a fluidics station 400, using the standard protocol EUkGEWS2v4 and scanned using an Agilent GeneArray Scanner.
After hybridization the microarray slides were washed using the standard protocol (Agilent Technologies, USA) and scanned on an Agilent microarray scanner.
This was generated using the standard dual-point VIBE T1-weighted Dixon sequence provided by the manufacturer on the scanner.
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