Skip to main content
. 2012 Dec 14;7(12):e50915. doi: 10.1371/journal.pone.0050915

Figure 2. Comparison of multi-beam striped-illumination TPLSM to standard single- and multi-beam TPLSM.

Figure 2

(a) 3D fluorescence images of spleen slices of chimera EGFP mice reconstituted with a non-fluorescent immune system using single-beam scanning PMT-TPLSM (SB-PMT), multi-beam scanning CCD-TPLM (MB-CCD) and SI-TPLSM (MB-SI). λexc = 920 nm, grid unit = 16.5 µm, grid unit of the cropped image = 2.0 µm. (b) 3D fluorescence images of the same regions within a popliteal lymph node after immunization with NP-CGG as recorded by MB-CCD-TPLSM versus MB-SI-TPLSM or by SB-PMT-TPLSM versus MB-SI-TPLSM. Follicular dendritic cells (FDC) are stained by CD21/CD35-Fab fragment-ATTO590 (magenta), while antigen-specific B1–8 cells express EGFP (green). λexc = 860 nm, grid unit = 15 µm. (c) 3D second-harmonic generation (SHG) signal images of the same region within a non-fluorescent lymph node as recorded by MB-CCD-, SB-PMT- and MB-SI-TPLSM. The SHG signal mainly originates from collagen fibers. λexc = 900 nm, grid unit = 10 µm (grid unit = 8 µm for MB-CCD-, MB-SI-TPLSM and 6 µm for SB-PMT-TPLSM). (d) 3D fluorescence images of acute cerebellum slices of CerTN L15 mice (expresses Cerulean and Citrine over the Thy1 cassette) recorded with the same set-ups. λexc = 850 nm, grid unit = 20 µm. All experiments were performed with the 20×, NA = 0.95 objective lens at z-step = 500 nm.