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3D Microscopy: Understanding the Give and Take on Instrument Performance to Enable Informed Decisions 

Biologists have a significant toolbox at their disposal when it comes to microscopically imaging 3D samples, such as organoids. From widefield microscopy to confocal, superresolution, multiphoton and lightsheet, each have their own set of pros and cons that must be carefully considered before making an informed choice on the most suitable to address your biological question. Often a correlative approach is required, applying several techniques to address the question from different perspectives. It is also crucial to consider the method of sample preparation and optimise each of the potential steps which can include fixation, permeabilisation, labelling and mounting. Further, the images generated by all techniques can be enhanced with post-processing techniques, such as deconvolution, which can enable or help to improve subsequent image analysis and interpretation.

This presentation aims to introduce each of the microscopy techniques that may be applicable for imaging 3D samples and highlight their relative performance attributes in terms of sample viability, speed, resolution, contrast and depth penetration – highlighting that each technique and instrument will come with trade-offs between these parameters1.

1 Jonkman, Brown, Wright, Anderson & North (2020) Tutorial: guidance for quantitative confocal microscopy. Nature Protocols 15:1585–1611

3D Microscopy: Understanding the Give and Take on Instrument Performance to Enable Informed Decisions

Biologists have a significant toolbox at their disposal when it comes to microscopically imaging 3D samples, such as organoids. From widefield microscopy to confocal, superresolution, multiphoton and lightsheet, each have their own set of pros and cons that must be carefully considered before making an informed choice on the most suitable to address your biological question. Often a correlative approach is required, applying several techniques to address the question from different perspectives. It is also crucial to consider the method of sample preparation and optimise each of the potential steps which can include fixation, permeabilisation, labelling and mounting. Further, the images generated by all techniques can be enhanced with post-processing techniques, such as deconvolution, which can enable or help to improve subsequent image analysis and interpretation.

This presentation aims to introduce each of the microscopy techniques that may be applicable for imaging 3D samples and highlight their relative performance attributes in terms of sample viability, speed, resolution, contrast and depth penetration – highlighting that each technique and instrument will come with trade-offs between these parameters1.

1 Jonkman, Brown, Wright, Anderson & North (2020) Tutorial: guidance for quantitative confocal microscopy. Nature Protocols 15:1585–1611

Experts
Dr. Graham Wright
Acting Director
Research Support Centre (RSC), A*STAR, Singapore

Dr. Graham Wright holds an interdisciplinary PhD in cell biology and physics from The University of Edinburgh and an MBA from the Warwick Business School at The University of Warwick. He is the Acting Director of A*STAR’s Research Support Centre (RSC) and the Director of the A*STAR Microscopy Platform (AMP). RSC offers everything from on-demand access to sophisticated scientific instruments and services, located within varied Technology Platforms, to a research consumables webstore, playing a crucial role in powering biomedical innovation in Singapore.

Dr. Graham has extensive experience, and a strong publication record, in applying advanced light microscopy to a wide range of biomedical research projects. Outside the laboratory, he is committed to science outreach and has featured as a judge on MediaCorp’s National Science Challenge TV show, presented a TEDx talk, had microscopy images displayed on the big screen in Times Square, New York and exhibited work at the National Museum of Singapore.

3D Microscopy: Understanding the Give and Take on Instrument Performance to Enable Informed DecisionsOct 09 2024
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