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For this montage, the microscope was moved along the medio-lateral axis and images obtained approximately every 100 µm (Figure 5B).
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As the microscope is moving relatively slowly while scanning (comparing to the circuit speed), this approach is sufficient for scanning speeds up to several millimeters per second (while recording both the number of fringes and phase) or several centimeters per second (while recording only the number of fringes).
The optical trap was utilized to hold a pathogen or microbead immobile while the microscope stage was moved to place the object at the desired location.
At t = 3 s, the microscope stage was moved to bring the flagellum underneath the trapped bead.
The microscope objective was moved ±250 nm in the z direction with 25 nm increments using PIFOC objective scanner (Physik Instrumente, Germany).
The microscope objective was moved from the top to the bottom of the coverslip field by field, and each new field was photographed for the measurement of alveolar mechanics.
During the measurements, the microscope stage was moved at a constant scanning rate of 200 μm/sby two orthogonal DC motors (M-112.IDG, Physik Instrumente) so that the encounter rate does not depend on the size of the observed species.
After this final assembly, the whole preparation was moved to the microscope.
After the proper operation of the device on a microscope had been ensured, the device was moved to a fluorescence detection setup.
Then the chamber was moved to the microscope stage and images were taken every 5 min for 3 to 5 h.
In this way the microscope objective was translated to move the focal plane along the axial direction as the retroreflector was moved the same distance to maintain the sample and the reference arms matched in length for interference.
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