Supplementary Figure 4: Fluorescence of unstained tissue vs. H&E stained tissue.
From: Nanoscale imaging of clinical specimens using pathology-optimized expansion microscopy
(A) Brightfield image of unstained formalin-preserved human lymph node tissue with invaded breast cancer. (B-E) Widefield fluorescent images of the same sample as in A in four fluorescent channels (listed below). (F) Bright field image of tissue from the same block as in A, stained with H&E. (G-J) Widefield fluorescent images of the sample of F in four fluorescent channels. Filters for each fluorescent channel (Semrock, Rochester, NY): B and G: excitation 628 ± 20 nm, emission 692 ± 20 nm; C and H: excitation 586 ± 10 nm, emission 628 ± 19 nm; D and I: excitation 482 ± 9 nm, emission 520 ± 14 nm; E and J: excitation 387 ± 6 nm, emission 409 nm and longer. Images from a given fluorescent channel are adjusted to have the same contrast. (K) Brightfield and widefield fluorescent images of tissue from the same block as in A, stained with H&E, after each step of ExPath processing. Images from a given fluorescent channel are adjusted to have the same contrast. Samples may be especially bright after the heat treatment and gelation steps due to the buffer compositions present. Scale bars: A-J, 500 μm; K, 40 μm (for yellow scalebars, post-expansion, physical size 184 μm; expansion factor: 4.58).
