For strains made in this study, two different clones with confirmed presence of tagged RSP6 in the flagella were analyzed. Cells were grown overnight to early logarithmic growth in liquid TAP media and imaged on a Nikon Eclipse TE 2000-E inverted microscope equipped with a Nikon dark-field oil condenser, a Nikon S Fluor 20 × 0.75 air objective and a red LED. Movies of 2,500 frames were recorded at a rate of 464 frames per second with a XiQ USB 3.0 superspeed camera (Ximea, model MQ013MG-ON). The plugin MTrack2 (https://imagej.net/MTrack2 ) was used to track cells in movies reduced to a frame rate of ~25 frames per second. For each clone 30–70 cells from at least three different glass slides were tracked. A measure for efficient swimming was calculated by dividing the path length of the cell (overall distance travelled) by its displacement (the shortest distance between the starting point and end point of the path). For beat frequency analysis, cells were tracked using MTrack2 in original movies (464 frames per second). For each track, the power spectra of the frame to frame displacement amplitude and displacement direction were multiplied to identify a peak that corresponds to average beat frequency. Statistical analysis was performed in Prism 7 (GraphPad, Inc). One-way ANOVA with Holm-Sidak test was used to determine statistical significance between strains.
S fluor 20 0.75 air objective
Manufactured by Nikon
The S Fluor 20 × 0.75 air objective is a high-performance microscope lens designed by Nikon. It features a 20x magnification and a numerical aperture of 0.75, optimized for use in air-based imaging applications.
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Lab products found in correlation
2 protocols using s fluor 20 0.75 air objective
1
Motility Analysis of Tagged RSP6 Strains
2
Motility Analysis of Tagged RSP6 Strains
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For strains made in this study, two different clones with confirmed presence of tagged RSP6 in the flagella were analyzed. Cells were grown overnight to early logarithmic growth in liquid TAP media and imaged on a Nikon Eclipse TE 2000-E inverted microscope equipped with a Nikon dark-field oil condenser, a Nikon S Fluor 20 × 0.75 air objective and a red LED. Movies of 2,500 frames were recorded at a rate of 464 frames per second with a XiQ USB 3.0 superspeed camera (Ximea, model MQ013MG-ON). The plugin MTrack2 (https://imagej.net/MTrack2 ) was used to track cells in movies reduced to a frame rate of ~25 frames per second. For each clone 30–70 cells from at least three different glass slides were tracked. A measure for efficient swimming was calculated by dividing the path length of the cell (overall distance travelled) by its displacement (the shortest distance between the starting point and end point of the path). For beat frequency analysis, cells were tracked using MTrack2 in original movies (464 frames per second). For each track, the power spectra of the frame to frame displacement amplitude and displacement direction were multiplied to identify a peak that corresponds to average beat frequency. Statistical analysis was performed in Prism 7 (GraphPad, Inc). One-way ANOVA with Holm-Sidak test was used to determine statistical significance between strains.
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