“White and Clear” (WaC): A tunable hydrophilic tissue clearing strategy for multi‐species 3D imaging
Abstract Optical tissue clearing (OTC) techniques have transformed three‐dimensional (3D) biological imaging, yet existing protocols often face limitations related to pigment persistence, reagent viscosity, or restricted applicability across tissues and species. Here, we introduce White and Clear (WaC), a tunable hydrophilic tissue clearing strategy developed from the CUBIC (clear, unobstructed brain imaging cocktails) workflow that integrates efficient delipidation, hydrogen peroxide‐based pigment removal, and customizable refractive index matching using 2,2′‐thiodiethanol (TDE). WaC provides rapid, versatile, and reproducible clearing across multiple organs and species while preserving morphology and immunoreactivity, and enabling deep 3D imaging with confocal and light‐sheet microscopy. WaC improves ease of use, versatility and clearing efficiency across samples from multiple species, including rodents, pigs, and humans; it also includes the capability for whole‐body clearing of small animals like mice; compatibility with 3D imaging techniques such as selective plane illumination microscopy (SPIM)/light‐sheet fluorescence microscopy (LSFM); the possibility of fine‐tuning clearing parameters for optimal results in different situations; and the avoidance of cumbersome synthesis or application procedures as well as the use of toxic reagents.
Authors
- R. J. Jiménez Riobóo (ORCID: https://orcid.org/0000-0002-9545-0172)
- Jorge Ripoll (ORCID: https://orcid.org/0000-0001-8856-7738)
- Daniel Sanderson (ORCID: https://orcid.org/0000-0002-5247-7471)
- María Victoria Gómez‐Gaviro (ORCID: https://orcid.org/0000-0002-8683-5150)
- Vicente Llorente (ORCID: https://orcid.org/0000-0003-3374-0008)
- Isabel Izquierdo‐Barba (ORCID: https://orcid.org/0000-0002-4139-4646)
- Rafael Samaniego (ORCID: https://orcid.org/0000-0002-3081-7332)
- Esther Bernaldo-de-Quirós (ORCID: https://orcid.org/0000-0001-8470-9882)
- Elena Fernandez-Cortes
- J Bueno Romero
- Rafael Correa‐Rocha
- Juan Miguel Gil‐Jaurena
- Manuel Desco
Institutions
- Universidad Complutense de Madrid (ES)
- Spanish National Centre for Cardiovascular Research (ES)
- Hospital General Universitario Gregorio Marañón (ES)
- Centro de Investigación Biomédica en Red de Salud Mental (ES)
- Centro de Investigación en Red en Enfermedades Cardiovasculares (ES)
- Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine (ES)
- Hospital Universitario Puerta de Hierro Majadahonda (ES)
- Centro de Investigación Biomédica en Red (ES)
- Instituto de Ciencia de Materiales de Madrid (ES)
- Universidad Carlos III de Madrid (ES)
Publication Details
- Journal
- Open Research (University of Surrey)
- Published
- 2026-06-29
- DOI
- https://doi.org/10.1002/viw.20260029
- Primary Topic
- Advanced Fluorescence Microscopy Techniques
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Fundación Mutua Madrileña
- Comunidad de Madrid
- Ministerio de Ciencia, Innovación y Universidades
- Centro de Investigación Biomédica en Red Enfermedades Cardiovasculares
- Instituto de Salud Carlos III
- European Regional Development Fund