Figure 2.
land-ExM labeling and anchoring strategies improve the signal of TREx and pan-ExM. (A) Workflow of land-pan-ExM, which only replaces the labeling strategy of pan-ExM with the labeling strategy of land-ExM. (B) land-TREx protein channel of U2OS cells, where proteins were labeled and anchored with NHS-biotin-MA. Scale bar: 5 µm in pre-expansion unit. Linear expansion factor: 7. (C) TREx protein channel of U2OS cells, where proteins were anchored with acryloyl-X SE and stained with Alexa Fluor 488 NHS ester. Scale bar: 5 µm in pre-expansion unit. Linear expansion factor: 7. (D) Bar chart comparing the signal-to-noise ratio of the protein channel in land-TREx and TREx. The signal-to-noise ratio is calculated as the average pixel value of the area with cells divided by the average pixel value of the area without cells in each image. Each bar represents the mean ± standard error of more than 20 cells. (E) land-TREx lipid channel of U2OS cells, where lipids were labeled by mCLING and anchored with NHS-biotin-MA. Scale bar: 5 µm in pre-expansion unit. Linear expansion factor: 7.0. (F) TREx lipid channel of U2OS cells, where lipids were anchored with acryloyl-X SE and stained with mCLING. Scale bar: 5 µm in pre-expansion unit. Linear expansion factor: 7.0. (G) Bar chart comparing the signal-to-noise ratio of the lipid channel of land-TREx and TREx. The signal-to-noise ratio is calculated as the average pixel value of the area with cells divided by the average pixel value of the area without cells in each image. Each bar represents the mean ± standard error of more than 20 cells. (H) land-pan-ExM protein channel of U2OS cells, where proteins were labeled and anchored with NHS-biotin-MA. Scale bar: 5 µm in pre-expansion unit. Linear expansion factor: 12.0. (I) Pan-ExM protein channel of U2OS cells labeled with Alexa Fluor 488 NHS ester. Scale bar: 5 µm in pre-expansion unit. Linear expansion factor: 12.0. (J) Bar chart comparing the signal-to-noise ratio of the protein channel in land-pan-ExM and pan-ExM. The signal-to-noise ratio is calculated as the average pixel value of the area with cells divided by the average pixel value of the area without cells in each image. Each bar represents the mean ± standard error of more than 20 cells. (K) land-pan-ExM lipid channel of U2OS cells, where lipids were stained following the workflow (A). Scale bar: 5 µm in pre-expansion unit. Linear expansion factor: 12.0. (L) Pan-ExM lipid channel of U2OS cells labeled with mCLING. Scale bar: 5 µm in pre-expansion unit. Linear expansion factor: 12.0. (M) Bar chart comparing the signal-to-noise ratio of the lipid (mCLING) channel in land-pan-ExM and pan-ExM. The signal-to-noise ratio is calculated as the average pixel value of the area with cells divided by the average pixel value of the area without cells in each image. Each bar represents the mean ± standard error of more than 20 cells. All images were taken with an Airyscan microscope.

land-ExM labeling and anchoring strategies improve the signal of TREx and pan-ExM. (A) Workflow of land-pan-ExM, which only replaces the labeling strategy of pan-ExM with the labeling strategy of land-ExM. (B) land-TREx protein channel of U2OS cells, where proteins were labeled and anchored with NHS-biotin-MA. Scale bar: 5 µm in pre-expansion unit. Linear expansion factor: 7. (C) TREx protein channel of U2OS cells, where proteins were anchored with acryloyl-X SE and stained with Alexa Fluor 488 NHS ester. Scale bar: 5 µm in pre-expansion unit. Linear expansion factor: 7. (D) Bar chart comparing the signal-to-noise ratio of the protein channel in land-TREx and TREx. The signal-to-noise ratio is calculated as the average pixel value of the area with cells divided by the average pixel value of the area without cells in each image. Each bar represents the mean ± standard error of more than 20 cells. (E) land-TREx lipid channel of U2OS cells, where lipids were labeled by mCLING and anchored with NHS-biotin-MA. Scale bar: 5 µm in pre-expansion unit. Linear expansion factor: 7.0. (F) TREx lipid channel of U2OS cells, where lipids were anchored with acryloyl-X SE and stained with mCLING. Scale bar: 5 µm in pre-expansion unit. Linear expansion factor: 7.0. (G) Bar chart comparing the signal-to-noise ratio of the lipid channel of land-TREx and TREx. The signal-to-noise ratio is calculated as the average pixel value of the area with cells divided by the average pixel value of the area without cells in each image. Each bar represents the mean ± standard error of more than 20 cells. (H) land-pan-ExM protein channel of U2OS cells, where proteins were labeled and anchored with NHS-biotin-MA. Scale bar: 5 µm in pre-expansion unit. Linear expansion factor: 12.0. (I) Pan-ExM protein channel of U2OS cells labeled with Alexa Fluor 488 NHS ester. Scale bar: 5 µm in pre-expansion unit. Linear expansion factor: 12.0. (J) Bar chart comparing the signal-to-noise ratio of the protein channel in land-pan-ExM and pan-ExM. The signal-to-noise ratio is calculated as the average pixel value of the area with cells divided by the average pixel value of the area without cells in each image. Each bar represents the mean ± standard error of more than 20 cells. (K) land-pan-ExM lipid channel of U2OS cells, where lipids were stained following the workflow (A). Scale bar: 5 µm in pre-expansion unit. Linear expansion factor: 12.0. (L) Pan-ExM lipid channel of U2OS cells labeled with mCLING. Scale bar: 5 µm in pre-expansion unit. Linear expansion factor: 12.0. (M) Bar chart comparing the signal-to-noise ratio of the lipid (mCLING) channel in land-pan-ExM and pan-ExM. The signal-to-noise ratio is calculated as the average pixel value of the area with cells divided by the average pixel value of the area without cells in each image. Each bar represents the mean ± standard error of more than 20 cells. All images were taken with an Airyscan microscope.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal