Correction: Visualizing cellular and tissue ultrastructure using Ten-fold Robust Expansion Microscopy (TREx)

Main text

Damstra HGJ, Mohar B, Eddison M, Akhmanova A, Kapitein LC, Tillberg PW. 2022.7 Visualizing cellular and tissue ultrastructure using Ten-fold Robust Expansion8 Microscopy (TREx). eLife 11:e73775. doi: 10.7554/eLife.73775.

Published 18 February 2022

In February 2022, we published our paper Visualizing cellular and tissue ultrastructure using Ten-fold Robust Expansion Microscopy (TREx), which introduced a simple and versatile approach for single-step ten-fold expansion of cells and tissue. We reported that the bisacrylamide concentration required for ten-fold expansion was different between labs in Ashburn and Utrecht, and suggested that the user test a range of bis concentrations to ensure ten-fold expansion. We have identified the source of this discrepancy to be a difference in the TEMED stock concentration between labs. As a result, the experiments in Figures 3E-G, 4 and 5 were performed with 1.5% TEMED and 90 ppm bisacrylamide, while the other experiments were performed with the reported TREx concentrations of 0.15% TEMED and 50 ppm bisacrylamide. After discovering this difference, we have replicated the experiments in Figures 3E-G, 4 and 5 with 0.15% TEMED and 50 ppm bis and obtained indistinguishable results. We have corrected the manuscript as follows:

#1 (Discussion)

Original text:

Therefore, we based our TREx recipe on recipe family D. The exact crosslinker concentration that produces 10-fold expanding gels was found to vary between labs (i.e., 50 µg/mL in Ashburn, VA, USA versus 90 µg/mL in Utrecht, the Netherlands, possibly due to differences in gelation chamber design), so we recommend that each lab test a range of crosslinker concentrations between 30 and 100 µg/mL using their choice of specimen, gelation chamber, and incubator.

Revised text:

Therefore, we based our TREx recipe on recipe family D. The exact crosslinker concentration that produces 10-fold expanding gels can vary due to differences in gelation chamber design, so we recommend that each lab test a range of crosslinker concentrations between 30–60 ppm using their choice of specimen, gelation chamber, and incubator.

#2 (Materials and Methods)

Original text:

TREx gelation solution contains 1.1 M sodium acrylate, 2.0 M acrylamide (AA), 50 µg/mL (for tissue slices and cultured cells prepared at Janelia), or 90 µg/mL (for cultured cells prepared at Utrecht University) N,N′-methylenebisacrylamide (bis), PBS (1×), 1.5 mg/mL APS, 1.5 mg/mL TEMED, and (optionally, for thick tissue slices) 15 µg/mL 4-hydroxy TEMPO (4HT, Sigma, 176141).

Revised text:

The TREx gelation solution contains 1.1 M sodium acrylate, 2.0 M acrylamide (AA), 50 ppm N,N’-methylenebisacrylamide (bis), PBS (1x), 1.5 ppt APS, 1.5 ppt TEMED, and (optionally, for thick tissue slices) 15 ppm 4-hydroxy TEMPO (4HT, Sigma, 176141).

#3 (Materials and Methods)

Original text:

For TREx, samples were treated with 100 µg/mL acryloyl-X SE (AcX) (Thermo Fisher, A20770) in PBS overnight at RT. TEMED and APS were added to monomer solution (1.5 mg/mL each) to produce gelation solution.

Revised text:

For TREx experiments shown in Figures 3E–G, 4 and 5, samples were treated with 100 μg/mL acryloyl-X SE (AcX) (Thermo Fisher, A20770) in PBS overnight at RT. In these experiments, TEMED and bis were used at a concentration of 15 ppt and 90 ppm, respectively.

Article and author information

Author details

  1. Mark Eddison

Publication history

  1. Version of Record published: November 29, 2022 (version 1)

Copyright

© 2022, Damstra et al.

This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

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  1. Hugo GJ Damstra
  2. Boaz Mohar
  3. Mark Eddison
  4. Anna Akhmanova
  5. Lukas C Kapitein
  6. Paul W Tillberg
(2022)
Correction: Visualizing cellular and tissue ultrastructure using Ten-fold Robust Expansion Microscopy (TREx)
eLife 11:e85169.
https://doi.org/10.7554/eLife.85169