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Histology Utilization in Biomedical ResearchGrace Jenson
May 5, 2017
Role of the histotechnician
• Biomedical
• Research (translation to human models)
• Direct involvement in sample collection and discussion• Identification of appropriate histological methods
Review of regenerative medicine
• Three interrelated approaches:• Rejuvenation
• Replacement
• Regeneration
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Stem cell 101
• Stem cell basics
• Embryonic vs adult stem cells
• Induced pluripotent stem cells
Stem Cells 101
https://www.youtube.com/watch?v=aAGaPH3s74g#action=share
https://www.rndsystems.com/resources/articles/differentiation-potential-induced-pluripotent-stem-cells
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Examples in application
• Wound healing• Matrix (tissue engineering)• Processing and fixation• Staining
• Sucrose gradient processing• Endogenous fluorescent tags
• Trace MRNA-lineage tracing
• Whole organ review (mega block)
• Electron microscopy• Resin embedded samples
Examples in application
• Wound healing• Matrix (tissue engineering)
• Processing and fixation
• Staining
Examples in application
• Sucrose gradient processing• Endogenous fluorescent tags
• Trace MRNA-lineage tracing
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http://www.cosmobio.co.jp/export_e/products/detail/products_sge_20070824.asp?entry_id=3423
https://www.nature.com/articles/ncomms12260
Examples in application
• Whole organ review (mega block)• Sliding microtome
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Examples in application
• Electron microscopy• Resin embedded samples
• Ultra structure
http://www.izw-berlin.de/electron-microscopy.html
Current and Future Trends
• Digital pathology
• Immunohistochemistry• Layered staining
• Modern fluorescent staining• Infrared/photon
• Mega blocks
Immunohistochemistry staining
• Triple staining/ multiple color fluorescence (current)• Alexa Fluor
• Infrared (LICOR) (future)• Proton emission
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http://circinterventions.ahajournals.org/content/7/4/560.figures-only
https://www.licor.com/bio/applications/tissue_sections/
Immunohistochemistry staining
• Triple staining/ multiple color fluorescence (current)
• Alexa Fluor
http://www.biocompare.com/9776-Antibodies/4478620-Alexa-Fluor-Labeled-Secondary-Antibodies-and-Streptavidins/
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Immunohistochemistry staining
• Infrared (LI-COR) (future)• Proton emission
https://www.google.com/url?sa=i&rct=j&q=&esrc=s&source=images&cd=&cad=rja&uact=8&ved=0ahUKEwjuwf-wk87TAhWD14MKHbu_DncQjhwIBQ&url=http%3A%2F%2Fwww.licor.com%2Fbio%2Fblog%2Freagents%2Firdye-reagents-reagents%2Fmicroscopy-with-near-infrared-fluorescence-for-outstanding-images%2Fattachment%2Fmicroscopy_img_d-2&psig=AFQjCNE7cQGt3sonvbBUbj2ZXvpzRWoqMg&ust=1493709226783553
Needs of research
• Whole organ review• Elimination of image stitching
• Efficiency and cost• Multiple staining
• Co-localization
• Limited samples/resources
• Vibratome• Neurology
• Versatility
• Troubleshooting
The ultimate goal of regenerative medicine
Healing from Within: The Promise of Regenerative Medicine
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Contact Information
References
Kang, J. Y. & Zheng, L. (2013). Rejuvenation: an integrated approach to regenerative medicine. Regenerative Medicine Research. Retrieved from https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4376090/pdf/40340_2013_Article_6.pdf
National Institutes of Health. (2016). Stem Cell Information. Retrieved from https://stemcells.nih.gov/info/basics/1.htm
Rivera, E.A. & Jayo, M. J. (2013). Histological evaluation of tissue regeneration using biodegradable scaffold seeded by autologous cells for tubular/hollow organ application. Methods in Molecular Biology. Retrieved from https://www.ncbi.nlm.nih.gov/pubmed/23494443
R & D Systems. (2017). Differentiation potential of induced pluripotent stem cells. R & D Systems: A Biotechne Brand. Retrieved from https://www.rndsystems.com/resources/articles/differentiation- potential-induced-pluripotent-stem-cells