For fluorescence microscopy, STING−/− MEFs were fixed with 2% paraformaldehyde in 75 mM phosphate buffer, permeabilized in 0.1% Triton X in PBS, washed with 2.5% FBS and 10 mM glycine in PBS, and then blocked with 10% FBS in PBS. Cells were then incubated with primary antibodies recognizing IRF-3 or NFκB (Santa Cruz sc-372) in 10% FBS in PBS. Subsequently, cells were washed and incubated with the Alexa 488-conjugated secondary antibody (Invitrogen). Cells were washed and incubated for 10 min with 0.5 μg/ml DAPI (4′,6-diamidino-2-phenylindole) (Sigma) and mounted onto glass slides using ProLong Antifade reagent (Invitrogen). Cells were imaged with the Leica SP8 X white light laser confocal microscope.
Sp8 x white light laser confocal microscope
Manufactured by Leica
The Leica SP8 X white light laser confocal microscope is a high-performance imaging system designed for advanced research applications. It features a white light laser that provides a continuous spectrum of excitation wavelengths, allowing for flexible and efficient fluorescence imaging. The system's core function is to capture high-resolution, three-dimensional images of biological samples through the use of confocal scanning technology.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using sp8 x white light laser confocal microscope
1
STING-Deficient MEF Immunofluorescence
2
Imaging and Tracing of Salivary Gland Ducts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fixed salivary glands were imaged using the Leica SP8 X White Light laser confocal microscope. Images were mainly obtained with a 20 × oil-immersed objective and processed with Fiji (ImajeJ2 v2.9.0/1.53t)46 (link). For 3d ductal reconstruction, images of the whole first lobe were acquired with a 20 × oil-immersed objective and then stitched together using the LAS software v2.8.0, Leica. The tracing of the ductal network was performed manually using a purpose-built graphical user interface, implemented in MATLAB (R2022b, The MathWorks, Inc., Natick, Massachusetts, USA). To make Supplementary Movie 1 the z-stack from long term live-imaging was stabilized during post-processing using the StackReg plugin47 (link) in Fiji.
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!