Shrinkage-mediated imaging of entire organs and organisms using uDISCO

Abstract

An imaging technique lets scientists peer through the skin of a whole mouse or rat to examine its organs after death. Ali Ertürk of the Ludwig Maximilians University of Munich in Germany and his colleagues created a technique called ultimate DISCO (uDISCO), which removes pigments and lipids from the tissues of dead animals using an organic solvent. This leaves the organs and skin intact but transparent, while preserving genetically encoded fluorescent proteins. The method revealed the nervous system of a mouse in stark detail. uDISCO also shrinks bodies by up to 65%, making it possible to image whole animals using light-sheet microscopy, which excels at imaging smaller samples. Nature Methods http://dx.

A Single Vector Platform for High-Level Gene Transduction of Central Neurons: Adeno-Associated Virus Vector Equipped with the Tet-Off System

“…Furthermore, by attaching the membrane-targeting and/or dendrite-targeting signals, the somatodendritic and/or axonal structures of the infected neurons were clearly visualized. Recently, many kinds of tissue-clearing methods have been developed, such as Scale [99, 100], CUBIC [101, 102], CLARITY [103, 104], SeeDB [105, 106], DISCO [107–109], etc. Although these techniques enable us to perform three-dimensional imaging with whole brain or thick-slice samples in large-scale, the intensity of fluorescent signal is critical for deep brain imaging and comprehensive analysis of neuronal structures [99].…”

A beginner’s guide to tissue clearing

“…Another fundamental limitation is that because lipids are removed, no direct lipid staining is possible. While initial versions of these methods had problems with the preservation of protein-based fluorescence beyond a few days, there are at least three examples of more recent protocols that have extended to several months the amount of time the fluorescent proteins can be imaged after clearing: FluoClearBABB (Schwarz et al, 2015), uDISCO (Pan et al, 2016), and embedding of a DBE-cleared sample into a resin (Becker et al, 2014). Another common disadvantage mentioned when discussing these methods is sample shrinkage, though recent improvements have either solved this issue (iDISCO+ (Renier et al, 2016)) or turned it into an advantage for specific applications that require the imaging of very large samples, like entire adult mice or young rats (uDISCO, Figure 1 (Pan et al, 2016)).…”

Three-dimensional, isotropic imaging of mouse brain using multi-view deconvolution light sheet microscopy

“…Here, we use an organic-solvent-based clearing method (uDISCO 24 ) to clarify the brain of a transgenic adult mouse that is labeled with Thy 1-green°uorescent protein (line M, Jackson Laboratory). The picture of cleared brain tissue is shown in supplementary¯gure S1 and compared to raw tissue.…”

“…However, LSFM is still challenging in this scenario for its axial resolution, which is typically 4 to 20 m, remaining insu±cient for three-dimensionally visualizing the single neurons across a large interactive area. [21][22][23][24][25] Therefore, the¯ne neural connections are not able to be revealed as what have been achieved by microoptical sectioning tomography (MOST) or sequential two photon excitation microscopy. [26][27][28][29] Furthermore, even for the optically-cleared organs, the light scattering as well as absorption still exists, largely deteriorating the quality from the depth of tissues.…”