Animals were immobilized with 10–100 mM sodium azide dissolved in M9, depending on developmental stage, mounted on a 2% agarose pad, and covered with a No. 1.5 coverslip. Spherical aberration was minimized using immersion oil matching. Z-stacks were acquired using a DeltaVision Core deconvolution imaging system (Applied Precision) with the InsightSSI light source; UApo 40×/1.35 NA oil immersion objective, PlanApo 60×/1.42 NA oil immersion objective, or UPlanSApo 100×/1.40 NA oil immersion objective (Olympus); the standard DeltaVision live cell excitation and emission filter set; and a Photometrics CoolSnap HQ2 CCD camera (Roper Scientific). Images were acquired and deconvolved with Softworx 5.5 (Applied Precision). Images are displayed as maximum intensity projections generated in Priism [32 (link)]. Images were pseudocolored and image brightness was linearly adjusted using Adobe Photoshop. IL2 dendrite lengths were measured using the Segmented Line tool in Fiji (NIH) [33 (link)]. To control for differences in head size, dendrite lengths are normalized to the distance from the cell body to the nose tip. p-values were generated using the Wilcoxon Rank-Sum (Mann-Whitney U) test in RStudio and adjusted using the Bonferroni correction.
Uapo 40 1.35 na oil immersion objective
Manufactured by Olympus
The UApo 40×/1.35 NA oil immersion objective is a high-performance objective lens designed for use in advanced microscopy applications. It offers a 40x magnification and a numerical aperture of 1.35, providing excellent image resolution and light-gathering capabilities. This objective is suitable for use with oil immersion techniques to achieve optimal optical performance.
Automatically generated - may contain errors
Lab products found in correlation
Photometrics coolsnap hq2 ccd camera, by Teledyne (2 mentions)
Uplansapo 100 1.40 na oil immersion objective, by Olympus (2 mentions)
Deltavision core deconvolution imaging system, by Cytiva (2 mentions)
Planapo 60 1.42 na oil immersion objective, by Olympus (2 mentions)
Softworx 5, by Cytiva (2 mentions)
1.4 na oil immersion objective, by Olympus (1 mentions)
3 protocols using uapo 40 1.35 na oil immersion objective
1
Microscopic Imaging of C. elegans Neurons
2
Laser Ablation Techniques for Sample Analysis
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We ablated samples with 13 to 15-nJ (Yb-fibre), 10-nJ (Ti:Sapphire), and 28-μJ (nanosecond) pulse energies, at 1–10 kHz repetition rate. We used 100 pulses in all cases except for ink ablations with the nanosecond laser, for which we used 5 pulses. We focused pulses with an Olympus UAPO 40×, 1.35 NA oil immersion objective for ink, bundle, and motoneuron ablations and an Olympus 100×, 1.4 NA oil immersion objective for motoneuron ablations. Where possible, laser power and pulse number were set to the minimum setting in which damaged could be observed.
3
Imaging Caenorhabditis elegans Embryos and Larvae
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Animals were prepared for imaging by mounting on a 2% agarose pad with a #1.5 coverslip. Larval animals were immobilized with sodium azide, young embryos (prior to twitching, ~1.5-fold) were simply imaged live, and older embryos were arrested to prevent movement as described in Cebul, McLachlan, and Heiman, 2020 (link). Early-stage embryos were acquired by dissecting adult hermaphrodites.
Image stacks were collected on a DeltaVision Core deconvolution imaging system (Applied Precision) with the InsightSSI light source; UApo 40×/1.35 NA oil immersion objective, PlanApo 60×/1.42 NA oil immersion objective or UPlanSApo 100×/1.40 NA oil immersion objective (Olympus); the standard DeltaVision live cell excitation and emission filter set; and a Photometrics CoolSnap HQ2 CCD camera (Roper Scientific). Image stacks were acquired and deconvolved with Softworx 5.5 (Applied Precision).
Representative images were generated as follows: Maximum intensity projections were created from contiguous optical sections in Fiji, then linearly adjusted for brightness in Adobe Photoshop. Multicolored images were generated by placing each channel in a separate false-colored screen layer in Photoshop. Adobe Illustrator was used to assemble final images.
Image stacks were collected on a DeltaVision Core deconvolution imaging system (Applied Precision) with the InsightSSI light source; UApo 40×/1.35 NA oil immersion objective, PlanApo 60×/1.42 NA oil immersion objective or UPlanSApo 100×/1.40 NA oil immersion objective (Olympus); the standard DeltaVision live cell excitation and emission filter set; and a Photometrics CoolSnap HQ2 CCD camera (Roper Scientific). Image stacks were acquired and deconvolved with Softworx 5.5 (Applied Precision).
Representative images were generated as follows: Maximum intensity projections were created from contiguous optical sections in Fiji, then linearly adjusted for brightness in Adobe Photoshop. Multicolored images were generated by placing each channel in a separate false-colored screen layer in Photoshop. Adobe Illustrator was used to assemble final images.
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