Light sheet microscopy, such as Selective Plane Illumination Microscopy (SPIM, [1]), reduces photo-toxic effects to a minimum. Due to the illumination of the sample in a thin volume around the focal plane no tissue outside the plane of interest is exposed and bleached. In addition, the fluorescence is collected with highly sensitive cameras. Combined with novel sample mounting techniques [2], SPIM has become a powerful technique for the long-term observation of fragile biological organisms [3, 4]. SPIM benefits from the latest camera technology and is therefore constantly improving in speed and sensitivity. Over the years it has become evident that light sheet microscopy is revolutionizing bio-imaging in several ways. Lately, we have shown that three-dimensional (3D) volumes can be imaged almost instantaneously using electrically tunable lenses (ETL) in SPIM [5]. This makes SPIM the fastest fluorescence microscopy technology for non-invasive 3D imaging. Even the dynamics of the beating zebrafish heart can be captured and the myo- and endocardial tissues as well as the blood can be visualized by 3D reconstruction [6].
Experiments have become possible that run at full speed using the best possible hardware, without being limited by the fragility of the sample. The speed advantage of the SPIM over other fluorescence technique can be utilized not only to image rapid events in developing tissues but also to record a large number of views for multi-view reconstruction. One key application of light-sheet technology includes the multi-dimensional imaging of the developing zebrafish larvae over extended periods of time [2]. I will give some examples of the unique capabilities of SPIM, especially for monitoring the development of the zebrafish heart [6] and the early endoderm [7].
References
[1] J. Huisken, et al., Science 305 (2004) 1007.
[2] A. Kaufmann, et al., Development 139 (2012) 3242.
[3] J. Huisken, D.Y.R. Stainier, Development 136 (2009) 1963.
[4] M. Weber, J. Huisken, Curr Opin Genet Dev 21 (2011) 566.
[5] F.O. Fahrbach, et al., Opt Express. 21(2013):21010.
[6] M. Mickoleit, et al., under review.
[7] B. Schmid, et al., Nat Commun 4 (2013) 2207.