Live neurons were imaged 2 days post-transfection in HBSS supplemented with 10 mM HEPES pH 7.4, 1× B27, 2 mM GlutaMAX, and 1 mM sodium pyruvate on an IX81 microscope with a FluoView FV1000 laser-scanning confocal unit operated using the FV10-ASW v3.01 software (Olympus). Fluorescence excitation was delivered using a 488-nm laser through a 40× 1.3-NA oil-immersion objective (Olympus). Emission was passed through a 530/40-nm emission filter. A single slice close to the bottom of the neuron was imaged using a 1-Airy pinhole setting. Images shown in Fig. 1b are representative images selected out of multiple fields of view.
1.3 na oil immersion objective
Manufactured by Olympus
Sourced in Japan
The 40× 1.3-NA oil-immersion objective is a high-performance lens designed for microscopy. It provides a magnification of 40× and a numerical aperture (NA) of 1.3, which allows for high-resolution imaging. This objective is designed to be used with immersion oil to improve the quality of the image.
Automatically generated - may contain errors
5 protocols using 1.3 na oil immersion objective
1
Confocal Imaging of Transfected Neurons
2
Fluorescent Microscopy Imaging Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fluorescent images were taken on a Zeiss Apotome 2 microscope with Axiovision software (Zeiss) or, for high magnification images, on a Nikon Ti system running Nikon Elements AR software or a Deltavision RT system running SoftWorx. Mouse time-lapse images were taken on a Deltavision Core system enclosed in an environment chamber maintained at 37°C and 5% CO2. Images were acquired using a 40× 1.3 NA oil immersion objective (Olympus), led light source (Applied Precision), and a CoolSNAP HQ2 CCD camera (Photometrics). Images were deconvolved in Huygens Professional (Scientific Volume Imaging) and processed using Image-J (FIJI). Chick time-lapses were taken on a Zeiss Cell Observer system enclosed in a chamber maintained at 37°C and 5% CO2. Images were acquired using a 40× 1.2 NA silicone immersion objective (Carl Zeiss), LED light source (Carl Zeiss), and a Flash4 v2 sCMOS camera (Hamamatsu). Images were deconvolved and processed using the Zen Blue software (Carl Zeiss).
3
Imaging Rat Cortical Neurons with Confocal Microscopy
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Images for Figure 2G were obtained as follows. Rat cortical neurons were transfected at 8 DIV and imaged 3–4 days post-transfection in HBSS supplemented with 10 mM HEPES pH 7.4, 1 × B27, 2 mM GlutaMAX, and 1 mM sodium pyruvate on an IX81 microscope with a FluoView FV1000 laser-scanning confocal unit. Fluorescence excitation was delivered using a 488 nm argon laser through a 40×/1.3-NA oil-immersion objective (Olympus, Japan). Emission was passed through a 530/40 nm emission filter. Z-sections were imaged using a 1-Airy pinhole setting and two-pass Kalman filtering. A maximum-intensity projection was generated from two to three sections spaced 2 μm apart.
4
Time-lapse Imaging of Embryo Slices
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Time-lapse imaging of embryo slices was performed using a Deltavision Core microscope system in a WeatherStation environmental chamber maintained at 37°C. (GE Healthcare). Imaging was limited to minimal exposure times (50–100 milliseconds) to detect low fluorescence levels (Das and Storey, 2014 (link); Das et al., 2012 (link)). Image acquisition was performed using an Olympus 40 × 1.3 NA oil immersion objective or an Olympus 40 × 1.25 NA silicone oil immersion objective, a solid stated LED light source and a CoolSnap HQ2 cooled CCD camera (Photometrics). Unless otherwise stated, 33–34 optical sections spaced 1.5 μm apart were acquired for each slice at 5–10 min intervals (exposure time 5–50 milliseconds for each channel, 512 × 512 pixels, 2 × 2 binning). For en face imaging of EB3-GFP comets, 5–8 optical sections spaced 0.5 μm apart were acquired at ~1.5–3.0 s intervals (exposure time of 150–200 milliseconds for the EB3-GFP comets). For the KillerRed and its control experiments, each slice was exposed to a total of 15 min of green light irradiation. Images were deconvolved using the SoftWorx image processing software. The position of the apical surface at each time point was monitored by acquiring a bright-field reference image at the middle of the z-stack.
5
Confocal Imaging of Transfected Neurons
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Live neurons were imaged 2 days post-transfection in HBSS supplemented with 10 mM HEPES pH 7.4, 1× B27, 2 mM GlutaMAX, and 1 mM sodium pyruvate on an IX81 microscope with a FluoView FV1000 laser-scanning confocal unit operated using the FV10-ASW v3.01 software (Olympus). Fluorescence excitation was delivered using a 488-nm laser through a 40× 1.3-NA oil-immersion objective (Olympus). Emission was passed through a 530/40-nm emission filter. A single slice close to the bottom of the neuron was imaged using a 1-Airy pinhole setting. Images shown in Fig. 1b are representative images selected out of multiple fields of view.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!