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Examples of Use

Single Color Time Lapse Imaging

Example of single color 48h time lapse imaging using the custom built light sheet microscope

The movie shows the temporal activation of Notch signalling across the retina from about 24hpf until 72hpf, using the TP1:VenusPEST transgenic zebrafish line. This zebrafish line contains a Notch responsive element (TP1) coupled to a Venus fluorophore which has been destabilized with a PEST sequence for fast degradation. During the movie one can see how Notch is first activated throughout the progenitor population and eventually becomes restricted within the Mueller glial cells.

The imaging was performed on our custom built upright light sheet microscope, using 515 nm excitation and acquiring volumes of  ~300um x 300um x 200um every 2.5 min. Shown are maximum intensity projections in three directions.

These images are taken in collaboration with Xana Almeida & Julia Oswald, Harris group.

Multi-Position / Large Area Light Sheet Imaging

Example of multi-position light sheet Imaging of GFP-labelled chick embryo

Multiple Stacks (stitched), ~2500x5600x500 px3 ≈ 7 Giga Voxel, FOV ~ 0.9 x 0.3 x 0.5 mm3

Chick Stitched Stack

in collaboration with Octavian Voiculescu

Multi Color Time Lapse Imaging

Example of multi color imaging of zebrafish eye using the custom built upright light sheet microscope

This movie shows division and migration of the retinal progenitors. Projections of Ptf1a inhibitory neurons (GFP+) can be seen as well.

Two transgenes were used: Ath5:gapRFP, where the Ath5 promotor drives the expression of membrane tagged RFP in retinal ganglion cells as well as in photoreceptors, horizontal and amacrine cells; and Ptf1a:cytGFP, where the Ptf1a promotor drives the expression of cytoplasmic GFP in amacrine and horizontal cells.

At the start of this movie, many of the cells are already labelled with RFP, whereas the GFP expression has just started in a few cells. As the retina develops, a wave of differentiation sweeps across the retina, as cells start to differentiate and migrate to their appropriate position. After 12 hours, the retinal ganglion cell layer is already established and we can observe some retinal ganglion cells axons exiting the eye and projecting towards the brain. At the same time, the amacrine cell layer has also been established. Towards the end of the movie we can see some Ptf1a+  horizontal cells migrating towards the edge of the retina, to establish the horizontal cell layer.

The imaging for both movies was performed on our custom built upright light sheet microscope, using sequential multi color excitation at 488 nm and 561 nm and acquiring volumes of  ~300umx300umx200um every 5 min. Shown are maximum intensity projections of the complete stack.

These images were acquired in collaboration with Xana Almeida, Harris group

Multi Photon Time Lapse Imaging

Example of two photon time lapse light sheet imaging

The movie shows a Ath5:gapRFP transgenic zebrafish retina. In this transgenic, the Ath5 promotor drives the expression of membrane tagged RFP in retinal ganglion cells as well as in photoreceptors, horizontal and amacrine cells. We can observe the axons of retinal ganglion cells exiting the eye and projecting towards the brain.

Shown are maximum intensity projections in three directions based on imaging stacks of ~ 300um x 300um x 200um stacks. The imaging was performed using 927 nm excitation every 2.5 min for more than 8 hours.

Project in collaboration with Xana Almeida, Harris group

Comparison 1 and 2 Photon Light Sheet Imaging

Comparison of 1 photon and 2 photon excitation using custom built light sheet microscope


The images show a 72hpf Ath5:gapRFP transgenic zebrafish retina. In this transgenic, the Ath5 promotor drives the expression of membrane tagged RFP in retinal ganglion cells as well as in photoreceptors, horizontal and amacrine cells. We can observe the axons of retinal ganglion cells exiting the eye and projecting towards the brain.

This 3d rendering of a single time point shows a comparison of single and two photon imaging on the custom built light sheet microscope. Depending on the actual requirements either excitation scheme can be used. Multi photon allows for better signal-to-noise whereas single photon allows to image a larger area, e.g to get a better overview.

Imaging performed in collaboration with Xana Almeida, Harris group

Multi Photon / Multi Color Light sheet Imaging

Multi Photon / Multi Color Light sheet Imaging of Drosophila Embryos

Maximum Intensity Projections of 1 Photon and 2 Photon images of stage 11 Drosophila embryos, immunostained for parasegments with Wingless.

For better clarity the images have been spectrally unmixed.

In collaboration with T. Finegan, Sanson group