Whole Organ Blood and Lymphatic Vessels Imaging (WOBLI)
Abstract Thin section histology is limited in providing 3D structural information, particularly of the intricate morphology of the vasculature. Availability of high spatial resolution imaging for thick samples, would overcome the restriction dictated by low light penetration. Our study aimed at opti...
Main Authors: | , , , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Nature Publishing Group
2018-01-01
|
Series: | Scientific Reports |
Online Access: | https://doi.org/10.1038/s41598-018-19663-w |
id |
doaj-6f54c7ca5d874fb798a1a7df2d40a6fe |
---|---|
record_format |
Article |
spelling |
doaj-6f54c7ca5d874fb798a1a7df2d40a6fe2020-12-08T04:56:00ZengNature Publishing GroupScientific Reports2045-23222018-01-01811910.1038/s41598-018-19663-wWhole Organ Blood and Lymphatic Vessels Imaging (WOBLI)Roni Oren0Liat Fellus-Alyagor1Yoseph Addadi2Filip Bochner3Hila Gutman4Shani Blumenreich5Hagit Dafni6Nava Dekel7Michal Neeman8Shlomi Lazar9Department of Biological Regulation, Weizmann Institute of ScienceDepartment of Biological Regulation, Weizmann Institute of ScienceDepartment of Life Sciences Core Facilities, Weizmann Institute of ScienceDepartment of Biological Regulation, Weizmann Institute of ScienceDepartment of Pharmacology, Israel Institute for Biological ResearchDepartment of Biomolecular sciences, Weizmann Institute of ScienceDepartment of Veterinary resources, Weizmann Institute of ScienceDepartment of Biological Regulation, Weizmann Institute of ScienceDepartment of Biological Regulation, Weizmann Institute of ScienceDepartment of Pharmacology, Israel Institute for Biological ResearchAbstract Thin section histology is limited in providing 3D structural information, particularly of the intricate morphology of the vasculature. Availability of high spatial resolution imaging for thick samples, would overcome the restriction dictated by low light penetration. Our study aimed at optimizing the procedure for efficient and affordable tissue clearing, along with an appropriate immunofluorescence labeling that will be applicable for high resolution imaging of blood and lymphatic vessels. The new procedure, termed whole organ blood and lymphatic vessels imaging (WOBLI), is based on two previously reported methods, CLARITY and ScaleA2. We used this procedure for the analysis of isolated whole ovary, uterus, lung and liver. These organs were subjected to passive clearing, following fixation, immunolabeling and embedding in hydrogel. Cleared specimens were immersed in ScaleA2 solution until transparency was achieved and imaged using light sheet microscopy. We demonstrate that WOBLI allows detailed analysis and generation of structural information of the lymphatic and blood vasculature from thick slices and more importantly, from whole organs. We conclude that WOBLI offers the advantages of morphology and fluorescence preservation with efficient clearing. Furthermore, WOBLI provides a robust, cost-effective method for generation of transparent specimens, allowing high resolution, 3D-imaging of blood and lymphatic vessels networks.https://doi.org/10.1038/s41598-018-19663-w |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Roni Oren Liat Fellus-Alyagor Yoseph Addadi Filip Bochner Hila Gutman Shani Blumenreich Hagit Dafni Nava Dekel Michal Neeman Shlomi Lazar |
spellingShingle |
Roni Oren Liat Fellus-Alyagor Yoseph Addadi Filip Bochner Hila Gutman Shani Blumenreich Hagit Dafni Nava Dekel Michal Neeman Shlomi Lazar Whole Organ Blood and Lymphatic Vessels Imaging (WOBLI) Scientific Reports |
author_facet |
Roni Oren Liat Fellus-Alyagor Yoseph Addadi Filip Bochner Hila Gutman Shani Blumenreich Hagit Dafni Nava Dekel Michal Neeman Shlomi Lazar |
author_sort |
Roni Oren |
title |
Whole Organ Blood and Lymphatic Vessels Imaging (WOBLI) |
title_short |
Whole Organ Blood and Lymphatic Vessels Imaging (WOBLI) |
title_full |
Whole Organ Blood and Lymphatic Vessels Imaging (WOBLI) |
title_fullStr |
Whole Organ Blood and Lymphatic Vessels Imaging (WOBLI) |
title_full_unstemmed |
Whole Organ Blood and Lymphatic Vessels Imaging (WOBLI) |
title_sort |
whole organ blood and lymphatic vessels imaging (wobli) |
publisher |
Nature Publishing Group |
series |
Scientific Reports |
issn |
2045-2322 |
publishDate |
2018-01-01 |
description |
Abstract Thin section histology is limited in providing 3D structural information, particularly of the intricate morphology of the vasculature. Availability of high spatial resolution imaging for thick samples, would overcome the restriction dictated by low light penetration. Our study aimed at optimizing the procedure for efficient and affordable tissue clearing, along with an appropriate immunofluorescence labeling that will be applicable for high resolution imaging of blood and lymphatic vessels. The new procedure, termed whole organ blood and lymphatic vessels imaging (WOBLI), is based on two previously reported methods, CLARITY and ScaleA2. We used this procedure for the analysis of isolated whole ovary, uterus, lung and liver. These organs were subjected to passive clearing, following fixation, immunolabeling and embedding in hydrogel. Cleared specimens were immersed in ScaleA2 solution until transparency was achieved and imaged using light sheet microscopy. We demonstrate that WOBLI allows detailed analysis and generation of structural information of the lymphatic and blood vasculature from thick slices and more importantly, from whole organs. We conclude that WOBLI offers the advantages of morphology and fluorescence preservation with efficient clearing. Furthermore, WOBLI provides a robust, cost-effective method for generation of transparent specimens, allowing high resolution, 3D-imaging of blood and lymphatic vessels networks. |
url |
https://doi.org/10.1038/s41598-018-19663-w |
work_keys_str_mv |
AT ronioren wholeorganbloodandlymphaticvesselsimagingwobli AT liatfellusalyagor wholeorganbloodandlymphaticvesselsimagingwobli AT yosephaddadi wholeorganbloodandlymphaticvesselsimagingwobli AT filipbochner wholeorganbloodandlymphaticvesselsimagingwobli AT hilagutman wholeorganbloodandlymphaticvesselsimagingwobli AT shaniblumenreich wholeorganbloodandlymphaticvesselsimagingwobli AT hagitdafni wholeorganbloodandlymphaticvesselsimagingwobli AT navadekel wholeorganbloodandlymphaticvesselsimagingwobli AT michalneeman wholeorganbloodandlymphaticvesselsimagingwobli AT shlomilazar wholeorganbloodandlymphaticvesselsimagingwobli |
_version_ |
1724391884041748480 |