Naive MΦ were infected using GFP expressing Mtb (MOI=1) for 4 h, washed and treated with either CD44TA-LIP or left untreated and incubated until 18 h post infection. The cells were then washed, plated onto coverslips, fixed in 3.7% paraformaldehyde, permeabilized using tween80-digitonin--BSA buffer. A specific mAb to LC3 (Cell signaling #3868) was used to stain autophagosomes using a Texas red conjugated secondary antibody (ThermoFisher Scientific, T-2767). IF images were acquired using a NIS element deconvolution software and N90 Nikon microscope. Colocalization was determined by scoring the localization of LC3B and GFP Mtb phagosomes of 25 MΦ in triplicate chambers per treatment group.
N90 microscope
Manufactured by Nikon
The N90 Nikon microscope is a laboratory-grade optical instrument designed for high-quality imaging and analysis. It features a sturdy construction, precise optics, and advanced illumination systems to provide clear, detailed observations of specimens. The core function of the N90 is to enable detailed examination and study of samples across a variety of scientific and research applications.
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Lab products found in correlation
2 protocols using n90 microscope
1
Autophagy Induction in Mtb-Infected Macrophages
2
Mtb Phagosome Colocalization Assay in MDSCs
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Naive MDSCs were infected using gfp-expressing Mtb (MOI of 1) for 4 h, washed and treated using either mab-B2 or isotype, and incubated at 18 h. Post infection, the cells were washed, plated onto coverslips, fixed in 3.7% paraformaldehyde, and permeabilized using Tween 80-digitonin-BSA buffer as described before (18 (link)). A specific mab to LC3 (Cell Signaling #3868) was used to stain gfpMtb phagosomes followed by Alexa fluor 555 conjugates. Confocal images were acquired using an N90 Nikon microscope equipped with a Metaview RT deconvolution software. Colocalization was determined by scoring gfpMtb phagosomes of 25 MDSCs in triplicate chambers per treatment group (18 (link), 21 (link)).
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