The retina has a very complex structure and working principle, since its task is to elaborate and transmit light signals to the optical nerve. Investigating its three dimensional structure in details represents a key point to understand its behavior. Lightsheet microscopy has been established as a powerful technique to reveal structures in thick biological specimens. Nevertheless, standard Selective Plane Illumination Microscopes (1) still rely on the embedding of the sample within agar gel or some other aqueous compound. This kind of solution limits the variety of samples that one can investigate under SPIM. To get rid of these limitations, we developed a version of inverted SPIM (2) which takes advantage of light sheet microscopy in a vertical geometry, where the specimen can be placed on standard petri dishes or coverslips, thus enhancing the portfolio of usable samples for SPIM. We performed imaging of transgenic mouse retina expressing standard fluorescent proteins (3), being interested on the retinal ganglion cell layer, to produce a 3D reconstruction of the development of dendrites. We were able to perform volume acquisition covering the entire thickness of the retina (approximately 300 microns) in less than 10 seconds with a large field of view, using a low NA detection objective. This technique combines the advantages of placing the sample in a standard substrate for microscopy with the speed of a widefield microscope, obtaining a resolution (with low NA objective) comparable to confocal microscopy. Moreover, 3D imaging can be achieved deeper into the sample than using standard microscopy techniques (4).(1) Ahrens M.B. et al. Nat. Methods 10 413-420 (2013).(2) Wu Y. et al. PNAS 108 (43) (2011).(3) Renvision project, FP7-ICT-2011-9 (NBIS) - Grant Agreement Number 600847.(4) Lavagnino Z. et al. Opt. Express 21 (5) (2013).