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FRET was carried out using two laser lines; firstly the 488 nm line of an Argon laser (Lasos) set at 6.1A using 5% laser power and a pinhole of 1. Secondly a 543 nm line from a HeNe laser (Lasos) using 100% laser power with a pinhole of 1.
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The detection system uses two laser lines that are projected onto the ground as probes.
The cantilever vibration was driven (purple dashed line) and detected (red solid line) optically using two laser beams.
Cy5-labeled microtubules and GFP-labeled proteins were visualized sequentially by switching between two laser lines and using a two-band filter set (Chroma technology).
Immunostained cells were then observed by confocal microscopy (LSM 510 Meta laser scanning microscope, Carl Zeiss) using a 63× objective lens and two laser lines for excitation with the following band pass settings: Argon 488 nm (band pass 505 530 nm), HeNe 543 nm (long pass 560 nm).
Samples were measured in both the radial breathing mode and tangential mode ranges using three different laser lines.
Confocal three-dimensional scans of 1024 × 1024 pixel images in a z-stack were taken with a Zeiss LSM 700 confocal microscope using three excitation laser lines (405 nm for DAPI, 488 nm for GFP, and 633 nm for ChAT) and imaged using Zen imaging software (Carl Zeiss MicroImaging GmbH).
Two laser lines were used, at 488 nm and 543 nm to excite the fluorochromes.
The original flow cytometers from the 1970s and 1980s used large water-cooled lasers to produce only one or two laser lines at a time.
Two laser lines at 488 and 561 nm were used for the excitation of GFP and pmRFP and the system was driven by IQ software (Andor Technology, UK).
For collecting the diffusions maps we used only one laser line (488 nm) to excite both eGFP and mRFP1.
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