diSPIM publications

Below is a list of some example research papers in which the diSPIM has been used for imaging biological specimens. A longer list can be found at http://dispim.org/media/publications

C. Elegans

Wu Y, Ghitani A, Christensen R, Santella A, Du Z, Rondeau G, Bao Z, Colon-Ramos D, Shroff H. Inverted selective plane illumination microscopy (iSPIM) enables coupled cell identity lineaging and neurodevelopmental imaging in Caenorhabditis elegans. Proc. Natl. Acad. Sci. USA 108 (43), 17708-17713, 2011. doi.org/10.1073/pnas.1108494108

The original paper describes the inverted SPIM configuration and its use for imaging C. Elegans development in the egg from two-cell stage to L1 larva every 2 seconds for 14 hours with no detectable phototoxicity.

Wu Y, Wawrzusin P, Senseney J, Fischer RS, Christensen R, Santella A, York AG, Winter PW, Waterman CM, Bao Z, Colon-Ramos DA, McAuliffe M, Shroff H. Spatially isotropic four-dimensional imaging with dual-view plane illumination microscopy. Nat. Biotechnol. 31, 1032-1038, 2013. doi.org/10.1038/nbt.2713

The paper describes the dual view enhancement of the iSPIM configuration to provide spatially isotropic imaging. Microtubules are tracked in live human endothelial cells in 2D, and 3D culture. Neurite outgrowth in C. Elegans brain development is tracked in 100 slice stacks captured every second for 910 time points.

Kumar A, Wu Y, Christensen R, Chandris P, Gandler W, McCreedy E, Bokinsky A, Colon-Ramos DA, Bao Z, McAuliffe M, Rondeau G, Shroff H. Dual-view plane illumination microscopy for rapid and spatially isotropic imaging. Nature Protocols 9(11), 2555-2573, 2014. https://doi.org/10.1038/nprot.2014.172

The microscope configuration at the LMCB is based on the design in the above paper.

Smith JJ, Kenny IW, Wolff C, Cray R, Kumar A, Sherwood DR, Matus DQ. A light sheet fluorescence microscopy protocol for Caenorhabditis elegans larvae and adults. Front. Cell Dev. Biol. 10.3389. https://doi.org/10.3389/fcell.2022.1012820

The authors use an index-matched curable hydrogel and FEP tubing to allow light sheet imaging of larval and adult C. Elegans.

Spheroids

Eismann B, Kreiger TG, Beneke J, Bulkescher R, Adam L, Erfle H, Herrmann C, Eils R, Conrad C. Automated 3D light-sheet screening with high spatiotemporal resolution reveals mitotic phenotypes. J. Cell Sci. 133 (11): jcs245043. doi.org/10.1242/jcs.245043

The authors developed a procedure for high throughput screening in live 3D cultures using the diSPIM by depositing cells on a grid of Matrigel droplets in a Greiner OneWell plate and allowing them to develop into spheroids. The diSPIM provided advantages in speed, axial resolution and low illumination dose. A special processing pipeline was developed to handle the 74 TB of data from the screen.

Bacterial Biofilms

Qin B, Fei C, Bridges AA, Mashruwala AA, Stone HA, Wingreen NS, Bassler BL. Cell position fates and collective fountain flow in bacterial biofilms revealed by light-sheet microscopy. Science 369 (6499), 71-77, 2020. https://doi.org/10.1126/science.abb8501

The authors used the diSPIM to image biofilm development in 3D from single cells every three minutes for 16 hours. The high time resolution and isotropic resolution improved the ability to track cells through a 3D mass of bacteria.

Zebrafish

Bhattacharya, D., Zhong, J., Tavakoli, S. et al. Strain maps characterize the symmetry of convergence and extension patterns during zebrafish gastrulation. Sci Rep 11, 19357 (2021). https://doi.org/10.1038/s41598-021-98233-z

The authors of this study undertook imaging of tissue strain within 450 μm stacks of gastrulating zebrafish embryos from 5 hpf every 2 minutes for around 12 hours. The advantages of the diSPIM were easier sample mounting and embryo survival.

Zaucker A, Mitchell CA, Coker HLE and Sampath K (2021) Tools to Image Germplasm Dynamics During Early Zebrafish Development. Front. Cell Dev. Biol. 9:712503. https://doi.org/10.3389/fcell.2021.712503

These researchers developed 3D-printed moulds for making agarose wells, allowing diSPIM imaging of the germplasm in multiple developing embryos during the first day of development, with a reference size of about 1 mm³ per embryo. Approximately 1 mm stacks were taken every 2.5 minutes for 5 hours.

Nogare DED, Natesh N, Vishwasrao HD, Shroff H, Chitnis AB (2020) Zebrafish Posterior Lateral Line primordium migration requires interactions between a superficial sheath of motile cells and the skin. eLife 9: e58251. https://doi.org/10.7554/eLife.58251

This study looked at the migration of the posterior lateral line primordium (PLLp). The diSPIM was used to generate isotropic resolution in 1% agarose embedded embryos 32 hpf to image cells that were basal to the PLLp and those that were overlying.