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Journal: STAR Protocols
Article Title: Protocol to annotate dendritic cell maturation types in vivo making use of lipid nanoparticle-based approaches
doi: 10.1016/j.xpro.2026.104395
Figure Lengend Snippet: Panel design For efficiency, staining mixes are prepared one day before the readout. These include mixes for experimental samples, single-stained cell controls, and single-stained bead controls ( C and 2D). Mixes are prepared without cells and added on the day of the readout. The optimized panel uses two sequential staining steps. (A) 96-well plate layout for single-stained beads (B). (B) 96-well plate layout for the staining for the single-stained cell controls, SS1: single-stained staining 1, SS2: singe-stained staining 2. (C) Panel overview and information on staining mixes for the experimental samples, single-stained beads and single-stained cell controls for conventional flow cytometry. (D) Panel overview and information on staining mixes for the experimental samples, single-stained beads and single-stained cell controls for FACS. We assign a number to each marker to simplify labeling of the wells for the single-stained controls. The total volume of the experimental staining mixes for experimental samples is determined by the number of samples to be stained. For single-stained controls, we always prepare the staining in a total volume of 200 μL. All staining mixes and control samples are then prepared according to the required antibody dilutions. For the channel corresponding to the LNPs, single-stained controls consist of cells from a mouse injected with LNPs. For single-stained beads, an antibody conjugated to a fluorochrome detected in the same channel need to be used.
Article Snippet: Add 500 μL of the 2× digestion buffer 1. c. Incubate the Eppendorf tube in a 37°C heat block and put the shaker on maximum for 20 min. d. Resuspend tissue every 10 min. e. Filter over a 5 mL
Techniques: Staining, Flow Cytometry, Marker, Labeling, Control, Injection