Egg chambers were first incubated under agitation during 1 h in Schneider medium supplemented with insulin, FBS, penicillin, streptomycin according to ref. 50 (link) and colcemid (0.25 μg ml−1). Chambers were then mounted directly in a droplet of Voltalef 10 s oil onto a 0.17 mm cover slips. Images were acquired with a Nikon microscope (Eclipse Ti2) coupled to a spinning disk module (Yokogawa CSU X1), a CCD camera (Coolsnap HQ2, Photometrics), and a 488 nm laser to excite GFP. Local ultraviolet-mediated inactivation of colcemid in a defined ROI (Fig. 5a,b ) was carried out with a pulsed ultraviolet-laser (355 nm, 400 ps, 20 kHz). Global ultraviolet-mediated inactivation (Fig. 5c ) was carried out with fluorescence HBO lamp.
Eclipse ti2
Manufactured by Yokogawa
Sourced in United States
The Eclipse Ti2 is a high-performance inverted microscope system designed for advanced imaging and analysis. It offers a modular and flexible platform for a wide range of applications in life science research and industrial analysis. The Eclipse Ti2 provides stable and precise control of the optical system, enabling high-quality image acquisition and seamless integration with various imaging techniques.
Automatically generated - may contain errors
4 protocols using eclipse ti2
1
Spatiotemporal Control of Microtubule Dynamics
2
Spinning Disk Confocal Imaging of DAPI-Stained Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cover slips were mounted on slides with VECTASHIELD Antifade Mounting Medium with 4′,6-diamidino-2-phenylindole (DAPI) (Vector Laboratories, H-1200). A Nikon Eclipse Ti2 equipped with a Yokogawa CSU-W1 Spinning Disk Confocal, 100×, 1.45 numerical aperture oil objective was used for imaging with a Nikon elements software. Ten z planes at 200-nm steps were captured. Images were deconvoluted in Nikon elements and then processed using ImageJ with FIJI plugin. Z planes were stacked, and minimum and maximum display values were set in ImageJ for each channel to properly view fluorescence. The plot profile function in ImageJ was used to generate line intensity graphs. Independent replicates were performed to confirm results.
3
Fluorescence Microscopy of Yeast Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells in the logarithmic phase were inoculated in SC medium and grown as described for the growth assay above. Fluorescence images were obtained with a Nikon Eclipse Ti2/Yokogawa CSU-X1 spinning-disk microscope with two Prime BSI scientific complementary metal oxide semiconductor (sCMOS) cameras (Teledyne Photometrics, USA), a LightHub Ultra laser light (Omicron Laserage, Germany), and an Apo total internal-reflection fluorescence (TIRF) ×100/1.49 oil lens (Nikon, Japan). Experiments were repeated at least three times. Representative images are shown in the figures.
4
FRAP Analysis of hTR in Cajal Bodies
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were grown on 35 mm glass bottom (#1.5) dishes (MatTek). Prior to each imaging session, cells were transferred to DMEM lacking phenol red and supplemented with HEPES. Imaging was performed on a Nikon Eclipse Ti2 equipped with a Yokogawa Electric CSU-W1 spinning disk module, and using a 100x objective (NA 1.35, Plan Apo). Image acquisition was performed using Nikon Elements software. A Tokai Hit stage-top incubator was used to control temperature, humidity, and CO2 levels. CBs with colocalizing hTR were identified, and a circle of 2 micron diameter around the Cajal body was bleached with 405 nm beam (Bruker). Five initial images were taken of each cell before bleaching. After bleaching, images were taken every 600 ms with a 100 ms exposure time for 60 s. Images were collected using two Andor iXon 88 Life EMCCD cameras connected by a twin-camera module (Cairn). Cells were illuminated with 488 nm light (for MS2-hTR) and 564 nm light (for mCherry-Coilin). Registration was corrected using Tetra Speck beads (Invitrogen) and images were flat-field corrected for the 488 nm channel using a solution of fluorescein (Model and Burkhardt, 2001) . FRAP analysis was performed in FIJI (Schindelin et al., 2012) using the FRAP Profiler plugin (http://worms.zoology.wisc.edu/ research/4d/4d.html). FRAP curves were fit to averages of the datasets in Prism using a one-phase association model.
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!