Live embryos were imaged at room temperature with two systems running the Volocity software: either a spinning disk confocal system (Perkin Elmer Ultraview), using an Olympus IX70 microscope with a planApo 60× 1.40 NA oil immersion objective; or another spinning disk confocal system (VT-QLC100 VisiTech International), using a Leica DM-IRB microscope with a HCX PL APO 63× 1.40 NA oil immersion objective. Running the μmanager software, we also alternatively used another spinning disk confocal system (CSU10 Yokogawa), using a Nikon Eclipse Ti-E microscope with a perfect focus system, equipped with a 60× 1.4 NA oil immersion objective. In all these scope options, control versus experimental groups were always kept within the same system to maintain consistency. Either 481 and 561 (Leica), or 488 and 561 (Olympus and Nikon), nm lasers were used to visualize GFP/Venus and RFP/mCherry in multiple fly lines. Using 0.5μm intervals, 20 slices were obtained every 30 seconds. All videos were captured with emission discrimination with corresponding filters.
Hcx pl apo 63 1.40 na
Manufactured by Leica camera
The HCX PL APO 63× 1.40 NA is a high-numerical aperture (NA) objective lens designed for Leica microscopes. It features a magnification of 63× and a numerical aperture of 1.40, which allows for high-resolution imaging and excellent light-gathering capabilities.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using hcx pl apo 63 1.40 na
1
Live Embryo Imaging Using Confocal Microscopy
2
Confocal Microscopy and Image Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunofluorescence images were acquired on the Leica TCS SP2 laser scanning confocal microscope with 488-, 543- and 633-nm laser lines mounted on an upright Leica DM RXA fluorescence microscope using a HCX PL APO 63×/1.40NA oil immersion objective. Acquisition parameters were as follows: 12-bit, 1,024 × 1,024 pixels, 2.6× digital zoom, 800 Hz scan speed, 4-line Kalman filtering, sequential (by line) channel imaging, and 10-slice z-stack of 5 μm. Immunohistochemically stained specimens were imaged in brightfield on an Olympus BX51 upright epifluorescent microscope using a UPLANFL 20×/0.5NA dry objective with the Infinity3 (Lumenera) CCD set to 1,936 × 1,456 pixels.
In FIJI image analysis software (http://fiji.sc/Fiji ), fluorescent z-stacks underwent background subtraction (1,000-pixel-radius rolling ball, no smoothing) and maximum-intensity z-projection. Brightness and contrast were adjusted by linear histogram stretching to enhance visibility. Any images to be compared to one another were processed in parallel. Similarly, brightfield images underwent linear histogram stretching to increase visibility uniformly across the image.
In FIJI image analysis software (
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!