Over the last few years, the Visikol portfolio of tissue clearing reagents have been used by over a thousand researchers from around-the-world and have been integral to dozens of publications. During this period of time, there have been many different tissue clearing techniques described in the literature as well as wide range of publications detailing methodologies for the imaging and labeling of 3D tissues as well as the analysis of the resulting 3D data sets. With all of these approaches and tools it can be overwhelming to try and adopt tissue clearing and 3D tissue imaging and thus Visikol has recently published an Ebook on tissue clearing which describes practical considerations for adopting tissue clearing and how to choose an approach, imaging modality and overall workflow.
Since the microscope was first introduced, researchers and clinicians have characrtized tissues through the use of ultra-thin two dimensional slices which are placed on microscope slides, stained and visualizied by a pathologist. However, this pradigm can be highly limited for complex tissues and complicated features such as vasculature and thus researchers in the last five-to-ten years have begun to emply tissue clearing approaches combined with fluorescent labeling and three-dimensional microscopy (e.g. confocal microscopy, light sheet microscopy) to image tissues in 3D. Of all the tools employed in a laboratory, tissue clearing and 3D tissue imaging can be one of the most challenging and complex as it involves the marriage of multiple disparite discplinces. Proper execution of tissue clearing approaches requires an indepth knowledge of fluorescent labeling, tissue processing, advanced microscopy, image processing and data processing.
While adopting tissue clearing can be challenging, at Visikol we have focused for the last few years on developing protocols, tools and reagents that are easy-to-use, rapid and inexpensive such that any researcher can quickly adopt them into their workflow. “To help support these efforts, we have developed a Tissue Clearing Ebook which provides a contextual overview to adopting any tissue clearing technique with feedback and lessons learned from hundreds of researchers,” described Visikol Product Manager Ian MacCloud. The Ebook is designed as a primer for adopting tissue clearing and gives researchers a break down of common pitfalls and key considerations to keep in mind during 3D imaging such as maximizing fluorescent signal, minimizing background fluorescence, when to use confocal or light sheet microscopy, which antibodies to use and how to process specific types of tissue (e.g. FFPE, frozen fresh) or 3D cell culture models (i.e. organoids, spheroids, microtisses).
“In working with lots of researchers, we have seen many of them get discouraged by tissue clearing in general as they see a 3D image of a whole mouse brain acquired with CLARITY, try to replicate the process in their lab and unfortunately have poor results. We really made the Ebook for these researchers as there are lots of mistakes researchers typically make in adopting tissue clearing such as jumping to a whole mouse brain before validating that their immunolabels can penetrate thick tissue or if they have enough fluorescent signal with thick samples,” described Visikol CSO Dr. Tom Villani. The goal of the tissue clearing Ebook is to provide a dense but short preview to tissue clearing that every researcher should review before adopting tissue clearing into their workflow. To download the Ebook click below: