Small intestines “Swiss rolls” were prepared, cut into 7 μm thick sections and fixed in cold 4% PFA. The sections were then incubated and stained with the following primary Antibodies: Hamster Anti-mouse CD11c (N418, BioLegend), Mouse Anti-mouse CD45.2 FITC (104, BioLegend)-FITC conjugated, Rat Anti-mouse Wnt4 (MAB4751, R&D Systems), followed by incubation with the Alexa Fluor and Cyanine dye series of secondary antibodies (Alexa Fluor® 488 AffiniPure F(ab’)₂ Fragment Donkey Anti-Mouse IgG (H+L), Allophycocyanin (APC) AffiniPure F(ab’)₂ Fragment Donkey Anti-Rat IgG (H+L), Cy™3 AffiniPure Goat Anti-Armenian Hamster IgG (H+L) supplied by Jackson ImmunoResearch. Nuclei were stained with DAPI and pseudo-colored. Detection of fluorescence was observed under Cy2, Cy3, and Cy5 filters on a Leica Inverted Microscope DMi8 S Platform and a Leica DM6000B microscope with an automated stage coupled with a Leica DFC350FX camera. Exposure time and fluorescence intensities were normalized to appropriate isotype control images. Each fluorescent channel was photographed separately, then merged and overlaid them atop the corresponding images.
Inverted microscope dmi8 s platform
Manufactured by Leica
The Leica Inverted Microscope DMi8 S Platform is a versatile microscope designed for various applications in life science research and cell biology. It features an inverted optical design, allowing for easy access to samples and direct observation of cell cultures. The DMi8 S Platform provides high-quality imaging capabilities, enabling users to visualize and analyze specimens with precision.
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Lab products found in correlation
2 protocols using inverted microscope dmi8 s platform
1
Immunofluorescence Staining of Mouse Intestine
2
In Situ Protein Interaction Analysis
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Archived rectosigmoid biopsy samples were from the SCOPE cohort at the University of California, San Francisco (UCSF). The SCOPE cohort is an ongoing longitudinal study of over 1,500 HIV-infected and uninfected adults followed for research purposes. The UCSF Committee on Human Research reviewed and approved the SCOPE study (IRB# 10–01218), and all participants provided written informed consent. PLA starter Duolink® In Situ Orange Starter Kit Mouse/Goat was obtained from Sigma Aldrich containing the PLA probes anti-goat MINUS and anti-mouse PLUS with the detection reagent DuoLink® Orange. Fluorescence for proximity ligation was observed under a Cy3 filter on a Leica Inverted Microscope DMi8 S Platform Formalin fixed paraffin embedded (FFPE) tissues were stained with primary antibodies for anti-LINGO2 Ab was raised against Goat (R&D Systems; AF3679) and anti-TFF3 raised against mouse (eBioscience; 5uclnt3) were used to detect the LINGO2 and TFF3 interactions in situ and for PLAs that followed the manufacturers instruction (Sigma). Technical controls omitting either one of the antibodies served as a negative control as no circular DNA oligonucleotides were able to be amplified.
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