Casper Chamber: Improving Drug Testing Efficiency in Lightsheet Microscopy

Session

Poster Session

Authors

Leonor Morgado (2), Anna Pezzarossa (1), Paulo Carriço (1), Ruth Diez del Corral (1), Davide Accardi (1)

Affiliations

1. Champalimaud Foundation
2. Instituto Gulbenkian de Ciência

Keywords

Lightsheet microscopy, pharmacological manipulation, zebrafish, neural development, imaging chamber, cell ablation.

Abstract text

Lightsheet microscopy achieves intrinsic optical sectioning of samples in a widefield-like setup, enabling gentle illumination, high spatial, and temporal resolutions. This has revolutionised imaging by allowing the observation of cellular processes at high speed over extended periods [1,2, 3]. The Zeiss Lightsheet Z1 system, specifically, images specimens within an imaging chamber containing approximately 16.5 mL of a suitable medium. While this medium can include drugs to monitor their effects on specimens in vivo, the cost and environmental impact of drugs can be limiting.

To overcome this difficulty, we introduce the Casper chamber, designed to use only 3.0 mL of the drug solution—5.5 times less than the Zeiss commercial chamber. Casper has undergone optical and biological validation, demonstrating minimal optical aberrations and supporting observations of living zebrafish larvae for at least 24 hours. As a proof-of-concept that the chamber can be used to image the effect of a particular drug, we employed the nitroreductase/metronidazole (NTR/Mtz) mediated ablation method to induce cell death. Comparable results were obtained with both the Zeiss commercial chamber and the Casper chamber, affirming the custom-made chamber's utility when reagents are limited or environmentally incompatible.

We also provide detailed instructions on manufacturing the Casper chamber, offering a versatile solution across diverse scientific fields with various model organisms.

References

[1] J. Huisken, “Optical Sectioning Deep Inside Live Embryos by Selective Plane Illumination Microscopy”, Science, vol. 305, no. 5686, pp. 1007–1009, Aug. 2004.

[2] J. Huisken and D. Y. R. Stainier, “Even fluorescence excitation by multidirectional selective plane illumination microscopy (mSPIM)”, Opt. Lett., vol. 32, no. 17, pp. 2608–2610, Sep. 2007.

[3] M. Weber and J. Huisken, “Light sheet microscopy for real-time developmental biology”, Curr. Opin. Genet. Dev., vol. 21, no. 5, pp. 566–572, 2011.