Unstained WT and tdTomato samples acquired in Aurora 5L (UV-V-B-YG-R)

Analysis of WT and tdTomato(+/+) mice samples in Cytek Aurora 5L: 1- Definition of the subset distribution of a tdTomato reporter in lung (stained samples). 2- Analysis of autofluorescence via optSNE based on WT unstained ("US") samples from Lung, skin and spleen. 3-Analysis of tdTomato mice-specific optSNE clusters in unstained tdTomato (+/+) samples, via comparison with WT samples.
Conclusion:
1- tdTomato expressed in resident Lung alveolar macrophages, abT cells and CD45- cells. 2-OptSNE resolves autofluorescence clusters with unique spectral signatures 3-OptSNE of WT and tdTomato unstained concatenated samples for all tissues allows the visualization of the tissue-specific tdtomato+ cell clusters exclusively found in tdTomato mice.
Notes:
File name meaning: -Tissue- LU = Lung. SP = Spleen. SK = Skin. -mice model- WT = wild type or tdTomato -/-. TOM = tdTomato reporter mice (tdTomato+/+). -replica- 1,2,3 = mice replica number. -sample type- S = stained with immunophenotyping panel. US = unstained samples. Stained Lung samples were analyzed via full spectrum unmixing with FCS Express, using the compensation control files. The description of the antibody conjugates used is shown in the attached picture (reagents table). The gating strategy used to identify lung resident subsets is attached. WT lung was used as a negative control for tdTomato expression all samples acquired after successful QC in Aurora 5L