Panoptic vDISCO imaging reveals neuronal connectivity, remote trauma effects and meningeal vessels in intact transparent mice

Abstract

Note: Manuscript videos are available at 'Supplementary material' section of BioRxiv and at the following link http://vdisco.isd-muc.de/ Abstract Analysis of entire transparent rodent bodies could provide holistic information on biological systems in health and disease. However, it has been challenging to reliably image and quantify signal from endogenously expressed fluorescent proteins in large cleared mouse bodies due to the low signal contrast. Here, we devised a pressure driven, nanobody based whole-body immunolabeling technology to enhance the signal of fluorescent proteins by up to two orders of magnitude. This allowed us to image subcellular details in transparent mouse bodies through bones and highly autofluorescent tissues, and perform quantifications. We visualized for the first-time whole-body neuronal connectivity of an entire adult mouse and discovered that brain trauma induces degeneration of peripheral axons. We also imaged meningeal lymphatic vessels and immune cells through the intact skull and vertebra in naïve animals and trauma models. Thus, our new approach can provide an unbiased holistic view of biological events affecting the nervous system and the rest of the body.

Hydrogen bound water profiles in the skin influenced by optical clearing molecular agents—Quantitative analysis using confocal Raman microscopy

“…In order to increase the probing depth, light‐focusing ability, spatial resolution of optical systems and image contrast, the optical clearing (OC) technique, allowing for control of tissue optical properties, has been proposed. Fundamentals and advances of OC technologies for in vitro and in vivo applications were recently reviewed in a number of publications . The influence of different OC agents (OCAs), for example, glycerol , glucose , dimethyl sulfoxide (DMSO) , scale , Sca/eS , PEG 400 and thiazone , was studied by various optical techniques such as OCT , MPT , RS , CRM and so on .…”

The mesoSPIM initiative: open-source light-sheet microscopes for imaging cleared tissue

“… We do not recommend having a 405-nm laser line unless it is absolutely needed -in general, the penetration depth even in cleared samples at this wavelength is very low 56 and it is recommended to use red labels instead of UV-excitable dyes.…”

“…The combination of benzyl alcohol and benzyl benzoate is among the oldest clearing techniques and was introduced in the early 20 th century by Spalteholz 8,68 . It forms the ancestor of a wide variety of clearing techniques based on dehydration, delipidation and index matching using organic solvents -including 3DISCO 6,7,69 , iDISCO 11 , iDISCO+ 70,71 , uDISCO 72 , vDISCO 56 and PEGASOS 73 . Compared to CLARITY and CUBIC, the refractive index of the employed organic solvents is usually >1.5, for example nD = 1.56 for BABB.…”

Use of high-refractive index hydrogels and tissue clearing for large biological sample imaging

“…However, change in size does not result in tissue disruption in the final cleared product and has minimal impact on outcomes 3,14,17 . In fact, increasing tissue volume can be used to increase resolution 18,19 , whereas decreasing the volume can be advantageous for large samples [20][21][22] . In addition to the impact of clearing protocols on tissue size, the chosen method can also alter the mechanical properties of the sample.…”