Analyses of lymphocyte subpopulations were performed on unfixed cell preparations with a fluorescence activated cell sorter (FACScan, Becton-Dickinson FACS Systems, Sunnyvale, California) and a panel of monoclonal antibodies (mAbs) to lymphocyte surface antigens (Ortho Diagnostic Systems: OKT3, OKT4, OKT8, OK-NK, OKB20, OKT26a, OK-DR; Thermo-Scientific, Rockford, IL, USA: TcRδ1 and δTcS1).
Preparations were observed with fluorescence microscope (DM5500, Leica) or a FV1000 (Olympus) confocal microscope.
We measured environmental lead concentrations in situ without sample preparation with a portable X-ray fluorescence analyzer InnovX Alpha (Innov-X Systems, Inc., Woburn, MA, USA) using soil mode.
Unbound antibody was removed by several 5 min rinses at room temperature with PBS prior to preparation for fluorescence microscopy.
In this protein, the targeted site was not the usual ATP binding site, but a new allosteric one, located in the C-terminal domain and identified with fluorescence experiments (May C. Morris, personal communication, manuscript in preparation).
Positions of vessels identified with fluorescence microlymphangiography could be also traced in fixed whole-mount ear preparations stained with vascular markers such as αSMA (Supplementary Fig. 2b c).
Sample preparation for fluorescence microscopy was done as per [ 26] with modifications.
The result of fluorescence intensity measurement with Fluorescence Spectrometer (Hitachi Ltd.
Fluorescence images were acquired with fluorescence microscope.
Fixed samples were analyzed with fluorescence microscopy.
