Figure 3.
land-ExM visualizes phase-separated and membrane organelles. (A–G) land-ExM protein images of membraneless phase separation structures. The proteins were labeled with NHS-biotin-MS and after gelation stained with streptavidin-Alexa Fluor 488. (A) land-ExM protein image of nucleoli in a U2OS cell. Red arrowheads indicate the fibrillar center (FC) or dense fibrillar component (DFC) of the nucleolus. Scale bar: 1 µm in pre-expansion unit. Linear expansion factor: 4.0. (B) land-ExM protein image of nuclear bodies of breast cancer cell, UCI082014. Red arrowheads indicate the nuclear bodies. Scale bar: 1 µm in pre-expansion unit. Linear expansion factor: 4.2. (C) land-ExM protein image of SGs of a U2OS cell treated with NaAsO2 for 20 min. The red arrowhead indicates a SG. Scale bar: 1 µm in pre-expansion unit. Linear expansion factor: 4.0. (D) land-ExM protein image of chromatin of a breast cancer cell. Scale bar: 500 nm in pre-expansion unit. Linear expansion factor: 4.2. (E) land-ExM protein image of NPCs of a breast cancer cell. Scale bar: 1 µm in pre-expansion unit. Linear expansion factor: 4.2. (F and G) land-ExM protein images of mitochondria and cytoskeleton of a U2OS cell. Scale bar: 1 µm in pre-expansion unit. Linear expansion factor: 4.0. (H–P) land-ExM lipid images of membrane structures. The lipids were labeled with mCLING-Atto647N. (H) land-ExM lipid image of breast cancer cell. Scale bar: 5 µm in pre-expansion unit. Linear expansion factor: 4.0. (I–M) magnified images of H showing different membrane structures: lipid vesicles (I), mitochondria (J), filopodia (K), nuclear invagination (L), and Golgi apparatus (M). Scale bar: 1 µm (I–M) in pre-expansion unit. (N) 3D land-ExM lipid image of a breast cancer cell after maximum intensity projection, showing the cell membrane. Color-coded by the z-dimension slices from bottom to top. Color bar: purple to white: 0–6 µm in pre-expansion unit. Scale bar: 5 µm in pre-expansion unit. Linear expansion factor: 4.0. (O and P) magnified images of N showing detailed structures of the cell membrane. Scale bar: 1 µm in pre-expansion unit. All images were taken with an Airyscan microscope.

land-ExM visualizes phase-separated and membrane organelles. (A–G) land-ExM protein images of membraneless phase separation structures. The proteins were labeled with NHS-biotin-MS and after gelation stained with streptavidin-Alexa Fluor 488. (A) land-ExM protein image of nucleoli in a U2OS cell. Red arrowheads indicate the fibrillar center (FC) or dense fibrillar component (DFC) of the nucleolus. Scale bar: 1 µm in pre-expansion unit. Linear expansion factor: 4.0. (B) land-ExM protein image of nuclear bodies of breast cancer cell, UCI082014. Red arrowheads indicate the nuclear bodies. Scale bar: 1 µm in pre-expansion unit. Linear expansion factor: 4.2. (C) land-ExM protein image of SGs of a U2OS cell treated with NaAsO2 for 20 min. The red arrowhead indicates a SG. Scale bar: 1 µm in pre-expansion unit. Linear expansion factor: 4.0. (D) land-ExM protein image of chromatin of a breast cancer cell. Scale bar: 500 nm in pre-expansion unit. Linear expansion factor: 4.2. (E) land-ExM protein image of NPCs of a breast cancer cell. Scale bar: 1 µm in pre-expansion unit. Linear expansion factor: 4.2. (F and G) land-ExM protein images of mitochondria and cytoskeleton of a U2OS cell. Scale bar: 1 µm in pre-expansion unit. Linear expansion factor: 4.0. (H–P) land-ExM lipid images of membrane structures. The lipids were labeled with mCLING-Atto647N. (H) land-ExM lipid image of breast cancer cell. Scale bar: 5 µm in pre-expansion unit. Linear expansion factor: 4.0. (I–M) magnified images of H showing different membrane structures: lipid vesicles (I), mitochondria (J), filopodia (K), nuclear invagination (L), and Golgi apparatus (M). Scale bar: 1 µm (I–M) in pre-expansion unit. (N) 3D land-ExM lipid image of a breast cancer cell after maximum intensity projection, showing the cell membrane. Color-coded by the z-dimension slices from bottom to top. Color bar: purple to white: 0–6 µm in pre-expansion unit. Scale bar: 5 µm in pre-expansion unit. Linear expansion factor: 4.0. (O and P) magnified images of N showing detailed structures of the cell membrane. Scale bar: 1 µm in pre-expansion unit. All images were taken with an Airyscan microscope.

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