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T cells from regional draining lymph nodes (LNs) of DED mice and normally sighted mice were analyzed for surface activation markers (CD69 and CD154), chemokine and cytokine receptors, and proliferation potential.
Cells were gated on CD4 positive cells and analyzed for surface expression of CD69.
At indicated time points, cells were harvested, counted, analyzed for surface expression of CD4 and CD25, and processed for CD25+ and CD25- T cell magnetic isolation.
Subsequently, cells were centrifuged, supernatant discarded, 200 µl FACS Buffer (BD Biosciences) added, and analyzed for surface marker expression by flow cytometric analysis.
GFP-positive cells were FACS sorted 24 hours post-transfection and analyzed for surface IFNAR1 and IFNAR2 expression using flow cytometry.
Randomly captured images were analyzed for surface area coverage, area distribution of individual segmented features or integrated fluorescence intensity with Image-Pro (ImagePro, Silver Spring MD, USA) or Metamorph software (MDS Analytical Technologies, Downingtown, PA, USA) [27].
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Live cells were identified by 7-aminoactinomycin D (7-AAD) exclusion and analyzed for surface-marker expression using FACS Calibur (BD Biosciences).
Live cells identified by 7-AAD exclusion were analyzed for surface-marker expression using FACSCalibur or sorted using a Dakocytomation Mo Flo high speed flowcytometer.
The materials were analyzed for several surface characteristics, including IR absorption, enzyme adsorption capacity, total surface area, cellulosic surface area, and cellulosic pore sizes.
Power and cooling systems were analyzed for Venus surface operation.
GFP positive cells were subsequently analyzed for cell surface MHC-I levels by flow cytometry using W6/32 anti-MHC-I antibody as described above.
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