Technological advances in three-dimensional (3D) reconstruction techniques have previously enabled paradigm shifts in our understanding of human embryonic and fetal development. Light sheet fluorescence microscopy (LSFM) is a recently-developed technique that uses thin planes of light to optically section whole-mount cleared and immunolabeled biologic specimens.
The advent of commercially-available light sheet microscopes has facilitated a new generation of research into protein localization and tissue dynamics at extremely high resolution. Our group has applied LSFM to study developing human fetal external genitalia, internal genitalia and kidneys. This review describes LSFM and presents our group's technique for preparing, clearing, immunostaining and imaging human fetal urogenital specimens. We then present light sheet images and videos of each element of the developing human urogenital system. To the extent of our knowledge, the work conducted by our laboratory represents the first description of a method for performing LSFM on the full human urogenital system during the embryonic and fetal periods.
Differentiation; research in biological diversity. 2019 Oct 10 [Epub]
Dylan Isaacson, Dylan McCreedy, Meredith Calvert, Joel Shen, Adriane Sinclair, Mei Cao, Yi Li, Todd McDevitt, Gerald Cunha, Laurence Baskin
Department of Urology, Northwestern University Feinberg School of Medicine, Chicago, IL, USA. Electronic address: ., Department of Biology, Texas A&M University, College Station, TX, USA., Histology and Light Microscopy Core, J. David Gladstone Institutes, San Francisco, CA, USA., CytomX Therapeutics, Inc. South San Francisco, CA, USA., Department of Urology, University of California, San Francisco, San Francisco, CA, USA., Department of Bioengineering and Therapeutic Sciences, J. David Gladstone Institutes, San Francisco, CA, USA; Institute of Cardiovascular Disease, J. David Gladstone Institutes, San Francisco, CA, USA., Department of Urology, University of California, San Francisco, San Francisco, CA, USA; Division of Pediatric Urology, University of California San Francisco Benioff Children's Hospital, San Francisco, CA, USA.
PubMed http://www.ncbi.nlm.nih.gov/pubmed/31634681