Using hydrogels in microscopy: A tutorial |
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Affiliation: | 1. School of Biology & Environmental Science, UCD Science Centre, Belfield, Dublin 4, Ireland;2. School of Biomolecular and Biomedical Science, UCD Science Centre, Belfield, Dublin 4, Ireland;1. Russell H. Morgan Dept. of Radiology and Radiological Science, Division of MR Research, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA;2. Cellular Imaging Section and Vascular Biology Program, Institute for Cell Engineering, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA;3. Dept. of Chemical & Biomolecular Engineering, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA;4. Dept. of Biomedical Engineering, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA;5. Dept. of Oncology, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA;1. Singapore University of Technology and Design, 8 Somapah Road, 487372 Singapore, Singapore;2. National University of Singapore, 21 Lower Kent Ridge Rd, 119077 Singapore, Singapore;1. Key Laboratory for Advanced Fuel and Chemical Propellant of the Ministry of Education, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China;2. Collaborative Innovative Center of Chemical Science and Engineering (Tianjin), Tianjin 300072, China;3. Haihe Laboratory of Sustainable Chemical Transformations, Tianjin 300192, China;1. Regenerative Medicine Program, Institute of Health and Biomedical Innovation, 60 Musk Ave, Kelvin Grove 4059 QLD, Brisbane, Australia;2. Cancer Program, Institute of Health and Biomedical Innovation, 60 Musk Ave, Kelvin Grove 4059 QLD, Brisbane, Australia;3. Faculty of Science and Engineering, Queensland University of Technology, Brisbane, Australia;4. Faculty of Health, Queensland University of Technology, Brisbane, Australia;5. Membrane Dynamics, Department of Biology, Technische Universität Darmstadt, Darmstadt, Germany;6. Department of Orthopaedics, University Medical Center Utrecht, Utrecht, The Netherlands;7. Orthopaedic Center for Musculoskeletal Research, Department of Orthopaedic Surgery, Julius-Maximilians-University Würzburg, Würzburg, Germany |
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Abstract: | Sample preparation for microscopy is a crucial step to ensure the best experimental outcome. It often requires the use of specific mounting media that have to be tailored to not just the sample but the chosen microscopy technique. The media must not damage the sample or impair the optical path, and may also have to support the correct physiological function/development of the sample. For decades, researchers have used embedding media such as hydrogels to maintain samples in place. Their ease of use and transparency has promoted them as mainstream mounting media. However, they are not as straightforward to implement as assumed. They can contain contaminants, generate forces on the sample, have complex diffusion and structural properties that are influenced by multiple factors and are generally not designed for microscopy in mind. This short review will discuss the advantages and disadvantages of using hydrogels for microscopy sample preparation and highlight some of the less obvious problems associated with the area. |
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Keywords: | 3D multiview imaging Light sheet microscopy Optical Projection Tomography Hydrogel agarose Live microscopy |
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