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For in vivo peptide challenge, mice were injected i.v. with 100 μg 2W1S peptide with 2.5 μg LPS, and secondary lymphoid tissues were harvested 4 hours later for flow cytometric analysis.
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Fresh leaves of 1-43 plants from most populations (274 individuals in total; Table 1) were later collected for flow cytometry analyses.
A total of 100 transplanted individuals were cultivated at the Botanical Garden of the University of Genova and later used for flow cytometry analyses at the Institute of Systematic Botany of the University of Zurich.
Single-cell suspensions were prepared from spleen and bone marrow and lysed for red blood cells (RBCs) using Ammonium-Chloride-Potassium (ACK) lysis buffer, and cells were washed with RPMI 1640 supplemented with 5% FCS and later used for flow cytometric analysis.
The solutions obtained are used to validate the general algorithm developed later for non-Newtonian flow in more general cross-sections.
Cells were harvested 20 h later for analysis by flow cytometry.
48 hours later cells were harvested for flow cytometry by trypsination.
(A ) Mice were i.v. immunized with SRBC-PKH26 or PBS control and analyzed 24 hr later by flow cytometry for PKH26 in gated DC subsets.
Mantle solidification and the drop in mantle heat flow occur slightly later for the inner model for two reasons: (1) the surface heat flow is lower than the other models in the first million years because the surface is hot, decreasing the upper mantle temperature jump; (2) tidal heating is initially moderate (∼0.1 TW) despite the mantle being mostly molten due to proximity to the star.
Five days later, spleen cells were harvested from adoptively transferred mice for flow cytometric analysis.
Viscera were either fixed in 4% formalin for immunohistochemistry, placed in RNA later (Life Technologies, ThermoFisher Scientific, Waltham, MA), or placed in phosphate-buffered saline for flow cytometry.
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