Light Sheet Microscopy & DRAQ probes

Since the development of Light Sheet Microscopy by the laboratory of Prof. Ernst Stelzer more than 10 years ago there have been many publications utilising DRAQ5™ and DRAQ7™.  A selection of these are presented below with brief notes on the samples that were interrogated.

Part of the power of Light Sheet Microscopy is the ability to optically section 3-dimensional structures while limiting out of focus light and its bleaching effect on fluorophores.  Aiding this, red-excited, far-red-emitting DRAQ™ probes benefit from less light scatter and have remarkable resistance to photo-bleaching (see: Martin et al., 2005) and chemical degradation (e.g. DRAQ7™ in multi-day cell health assays).

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Verveer, P. J., Swoger, J., Pampaloni, F., Greger, K., Marcello, M., & Stelzer, E. H. (2007). High-resolution three-dimensional imaging of large specimens with light sheet–based microscopy. Nature methods, 4(4), 311.

• GFP-MDCK cysts and BxPC-3 spheroids counterstained with DRAQ5™

Keller, P. J., & Stelzer, E. H. (2010). Digital scanned laser light sheet fluorescence microscopy. Cold Spring Harbor Protocols, 2010(5), pdb-top78.

• BxPC-3 spheroid: counterstained with DRAQ5™

Ejsmont, R. K., Sarov, M., Winkler, S., Lipinski, K. A., & Tomancak, P. (2009). A toolkit for high-throughput, cross-species gene engineering in Drosophila. Nature methods, 6(6), 435.

• Drosophila embryo: anti-GFP stained and counterstained with DRAQ5™

Lorenzo, C., Frongia, C., Jorand, R., Fehrenbach, J., Weiss, P., Maandhui, A., ... & Lobjois, V. (2011). Live cell division dynamics monitoring in 3D large spheroid tumor models using light sheet microscopy. Cell division, 6(1), 1.

• Capan-2 cell spheroid: counterstained with DRAQ5™

Pampaloni, F., Ansari, N., & Stelzer, E. H. (2013). High-resolution deep imaging of live cellular spheroids with light-sheet-based fluorescence microscopy. Cell and tissue research, 352(1), 161-177.

• T-47D tumour spheroid: counterstained with DRAQ5™ & MitoView™ Green (mitochondria)
• BxPC-3 spheroid: counterstained with DRAQ5™

Schmitz, A., Fischer, S. C., Mattheyer, C., Pampaloni, F., & Stelzer, E. H. (2017). Multiscale image analysis reveals structural heterogeneity of the cell microenvironment in homotypic spheroids. Scientific Reports, 7, 43693.

• T-47D tumour spheroid: counterstained with DRAQ5™ and cleared with BABB

Glaser, A. K., Reder, N. P., Chen, Y., McCarty, E. F., Yin, C., Wei, L., ... & Liu, J. T. (2017). Light-sheet microscopy for slide-free non-destructive pathology of large clinical specimens. Nature biomedical engineering, 1(7), 0084.

• Prostate core biopsy: cleared with X-CLARITY and stained with DRAQ5™ and eosin Y ("D&E"; see Elfer, et al. PloS one, 11(10), e0165530.)

Lawson, P. J., Hu, B., Fasy, B. T., Wenk, C., & Brown, J. Q. (2018, March). Quantifying prostate cancer morphology in 3D using light sheet microscopy and persistent homology (Conference Presentation). In Diagnosis and Treatment of Diseases in the Breast and Reproductive System IV (Vol. 10472, p. 1047209). International Society for Optics and Photonics.

• Biobanked Prostate-biopsies: stained with DRAQ5™ and eosin Y ("D&E"; see Elfer, et al. PloS one, 11(10), e0165530.)

Rieckher, M., Psycharakis, S. E., Ancora, D., Liapis, E., Zacharopoulos, A., Ripoll, J., ... & Zacharakis, G. (2018). Demonstrating Improved Multiple Transport‐Mean‐Free‐Path Imaging Capabilities of Light Sheet Microscopy in the Quantification of Fluorescence Dynamics. Biotechnology journal, 13(1), 1700419.

• T-47D tumour spheroid: imaged following 24h. incubation with 1.5 µM DRAQ7™ to label dead cells in the intact 3D structure

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