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Ask the Experts 03

Ask the Experts

Welcome to Ask the Experts, your one-stop portal for life science microscopy queries. Join our experts on live webinars to discuss trending topics, or contact our experts at any time with your questions and challenges. Missed a webinar? No problem, you can listen again. Don’t see a topic you’d like to discuss? Use our feedback form or contact our experts directly for help.

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Tissue Optical Clearing Imaging: From In Vitro to In Vivo

Tissue Optical Clearing Imaging: From In Vitro to In Vivo

Experts
Dr. Dan Zhu
Professor
Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Wuhan, China
Biomedical optical Imaging, as a powerful tool has been applied for observing biomedical tissue structural and functional information with high resolution and contrast unattainable by any other method. However, the high scattering of turbid biological tissues limits the penetration of light, leading to strongly decreased imaging resolution and contrast as light propagates deeper into the tissue. Fortunately, novel tissue optical clearing technique provide a way for reducing the scattering of tissue and improving the optical imaging quality. This presentation will introduce our progress from in vitro and in vivo of tissue optical clearing imaging, including developing in vitro optical clearing methods, such as FDISCO and MACS. Meanwhile, we will also demonstrate in vivo skull/skin optical clearing window for imaging structural and functional of cutaneous / cortical vascular and cells, also manipulating cortical vasculature.
3D Microscopy: Understanding the Give and Take on Instrument Performance to Enable Informed Decisions

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

Experts
Dr. Graham Wright
Acting Director
Research Support Centre (RSC), A*STAR, Singapore
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.
Advances in 3D Optical Imaging Technologies: An Overview

Advances in 3D Optical Imaging Technologies: An Overview

Experts
Mr. Srivats Hariharan
Manager, Applications and Marketing
Olympus Singapore
With rapid development in fluorescent proteins, synthetic fluorochromes, and digital imaging, advanced 3D imaging technologies are now available to investigators to provide critical insights into the fundamental nature of cellular and tissue functions. 3D and 4D imaging systems have become very common tools among biologists. However, there are several technical challenges and limitations in performing successful 3D and 4D imaging. Olympus has developed a wide range of 3D imaging microscopes to overcome these challenges and to satisfy the requirements of researchers across different disciplines.
Investigating Spheroid Architecture Using the FV3000 Confocal Microscope

Investigating Spheroid Architecture Using the FV3000 Confocal Microscope

Experts
Ms. Gency Gunasingh
Research Assistant
The University of Queensland Diamantina Institute
Phenotypic tumour heterogeneity arising due to differentially cycling cell populations has been implicated in increased therapy resistance. This phenomenon cannot be assessed in adherent cell culture, where microenvironmental conditions are homogeneous. Thus, we utilise melanoma spheroids to model the 3D tumour microenvironment including the extracellular matrix (ECM) and study spheroid structure, necrotic region, individual cell arrangement within and gene expression patterns. We achieve this by exploiting the fluorescence ubiquitination cell cycle indicator (FUCCI) system to monitor cell cycle stages as a surrogate marker for phenotypic tumour heterogeneity, tissue clearing and confocal microscopy using FV3000.
Prostate Cell Lineage Hierarchy and Plasticity

Prostate Cell Lineage Hierarchy and Plasticity

Experts
Dr. Dong Gao
Principal Investigator
Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences
Prostate cancer is one of the most common cancers worldwide and also the second leading cause of cancer-related death in males in Western countries. Although the majority of human primary prostate cancers have a luminal phenotype, both basal cells and luminal cells can serve as cellular origins of prostate cancer in model systems. However, the stem cell-like plasticity of defined prostate epithelial cells and the cellular origin of prostate cancer under physiological conditions have not been identified. Recently, prostate basal and luminal cell populations were both shown to be self-sustaining, and both cell types could initiate prostate cancer. However, the oncogenic transformation of basal cells requires basal to luminal cell transition. In addition, luminal cells were shown to have greater tendency to be the cells of origin for prostate cancer in some contexts.
3D Segmentation for Fluorescence Images: From Qualitative to Quantitative

3D Segmentation for Fluorescence Images: From Qualitative to Quantitative

Experts
Dr. Yu Weimiao
Head of Computational & Molecular Pathology Lab (CMPL) Agency of Science
Technology and Research
Cells are 3D functional elements in biology science and they are actively moving to perform their functions. Collective cell migration is appreciated as an important model for the understanding of the mechanism governing the cell movement in Vivo and in Vitro. It is a highly kinetic process involved in immune response, wound healing, tissue development and cancer metastasis. Recent decades have seen the fast development of various optical imaging techniques with excellent spatial-temporal resolution, dimensionality and scale. The generation of novel probes have also allowed us to acquire the movies of migrating cells with specific proteins/molecules. However, we lack of advanced solution to analyse such high-content and highly dynamic images/videos.
An In Vitro System for Evaluating Anticancer Drugs Using Patient-Derived Tumor Organoids

An In Vitro System for Evaluating Anticancer Drugs Using Patient-Derived Tumor Organoids

Experts
Dr. Motoki Takagi
Professor
Medical-Industrial Translational Research Center, Fukushima Medical University
Patient-derived tumor organoids (PDOs) represent a promising preclinical cancer model that better replicates disease, compared with traditional cell culture models. We have established a novel series of patient-derived tumor organoids (PDOs) from various types of tumor tissues from the Fukushima Translational Research Project, which are designated as Fukushima (F)-PDOs. F-PDOs could be cultured for >6 months and formed cell clusters with similar morphologies to their source tumors. Comparative histological and comprehensive gene-expression analyses also demonstrated that the characteristics of PDOs were similar to those of their source tumors, even following long-term expansion in culture. In addition, suitable high-throughput assay systems were constructed for each F-PDO in 96- and 384-well plate formats.
Study the Function of Stromal Cells through Intestinal Organoid Co-Culture Technology

Study the Function of Stromal Cells through Intestinal Organoid Co-Culture Technology

Experts
Dr. Ningbo Wu
Associate Professor
Shanghai Institute of Immunology, Shanghai Jiao Tong University School of Medicine
For a long period of time, intestinal mesenchymal stromal cells have been considered as a relatively simple and homogeneous group of cells. With the help of single cell transcriptomics studies, it has now been clear that these cells are quite complex and heterogeneous. However, the detailed cellular and molecular mechanisms that regulate the function of these cells remains poorly understood. Therefore, the ability to perturb and evaluate the function of these stromal cells is critical to the understanding of intestinal stem cell niche and the etiology of the inflammatory bowel diseases and colitis associated colorectal cancer.
NoviSight™ Demonstration: 3D Image Analysis and Statistical Software for Organoids and Spheroids

NoviSight™ Demonstration: 3D Image Analysis and Statistical Software for Organoids and Spheroids

Experts
Hiroya Ishihara
Applications Scientist
Olympus Technologies Asia, Olympus Corporation
Three-dimensional cell culture models such as patient-derived organoids (PDO) and spheroids have increased in popularity because they can provide a 3D microenvironment that more closely reproduces in vivo conditions compared to 2D monolayer culture. Phenotypic and functional heterogeneity arise among cancer cells within the same tumor because of genetic change, environmental differences and reversible changes in cell properties. Therefore, evaluation of cell-specific responses is important for accurate prediction of drug efficacy and kinetics in vivo.
Depth Matters: Transforming Biology for More Realistic and Meaningful Pursuits

Depth Matters: Transforming Biology for More Realistic and Meaningful Pursuits

Experts
Dr. Gowri Balachander
Research Fellow, Translation Mechanobiology lab, National University of Singapore
Improvements to in vitro three-dimensional (3D) models are making them increasingly better at mimicking in vivo-like cellular behavior. Every tissue presents a distinct microenvironment with a unique blend of biochemical and biophysical components that dictate cellular behavior. Recreation of critical features of tissues that nurtures recapitulation of in vivo-like cellular behavior is the essence of an effective 3D model. In this webinar, through specific examples of 3D models for tissue development and cancer, we will revisit the fundamental principles of designing 3D models that can effectively recapitulate critical features of the tissues in vitro and applications of such models in mechanistic studies and drug testing. Our work also highlights the importance of 3D imaging systems, such as laser scanning confocal microscopes, which are necessary for such work.

Three-Dimensional High-Throughput Image Analysis of Patient-Derived Organoids and Spheroids

Experts
Hiroya Ishihara
Applications Scientist
Olympus Technologies Asia, Olympus Corporation
Organoids and spheroids can more faithfully reproduce in vivo conditions, and imaging-based analysis can monitor cell-specific responses with high spatial resolution. Therefore, we have been developing imaging-based three-dimensional analysis and drug evaluation methods using patient-derived cancer organoids and spheroids.

Digital Image Processing Part 2: Advanced Image Processing Filter

Experts
Heiko Gäthje
Senior Trainer, Training Academy
Olympus Europa
In our first webinar on this topic, we learnt that images taken with a light microscope are never quite a perfect reflection of our samples. There are always sources of error, and as we can only minimize such errors, the raw data of an imaging experiment will often need some digital image processing before final image analysis.
ICSI—Past, Present & Future

ICSI—Past, Present & Future

Experts
Dr. Hrishikesh Pai
Medical Director, Bloom IVF Group
In this webinar, Dr. Pai will be sharing on what’s new in the management of male infertility with special focus on Intra Cytoplasmic Sperm Injection (ICSI). He will be describing the latest diagnostic modalities in male infertility especially sperm DNA fragmentation, genetic testing, cryotozoospermia and microsurgical TESA in cases of non-instructive azoospermia.

Microscope Objectives—Where the Magic Happens

Experts
Klaus Willeke
Product Marketing Manager
Lauren Alvarenga
Product Manager, Life Science Microscopy
In this webinar, Lauren and Klaus would like to emphasize the importance of good optics in a complex system like a microscope. They will discuss different aspects which are important for the final image quality.

Microscope Objectives—Where the Magic Happens

Experts
Ganesh Kadasoor
Application Specialist at Olympus Medical Systems India Ltd
Wei Juan Wong
Application Specialist
Olympus Soft Imaging Solutions
In this webinar, Olympus microscope experts Ganesh and Wei Juan discuss the importance of good optics in a complex microscope system and what optical features are important for high final image quality.
High-Content Screening: Customized Analysis Made Easy

High-Content Screening: Customized Analysis Made Easy

Experts
Manoel Veiga
Application Specialist, Life Science Research
Olympus Soft Imaging Solutions
Shohei Imamura
Strategic Project Manager, Regenerative Medicine and Drug Discovery Solutions
In this webinar, Manoel and Shohei, our deep learning and high-content screening experts, will introduce the scanR system’s assay builder. The scanR system is Olympus’ dedicated high-content screening platform with a unique sample navigation approach and analysis inspired by flow cytometry.
Live Cell Super Resolution Imaging: The Big Picture of Small Things

Live Cell Super Resolution Imaging: The Big Picture of Small Things

Experts
Chunsong Yan
Business Development Manager, Life Science
Olympus Australia & New Zealand
Lauren Alvarenga
Product Manager, Life Science Microscopy
Stefan Marawske
Senior Sales Specialist, High-End Life Science Systems
Olympus Europa
In this webinar, Olympus imaging experts Stefan, Lauren, and Chunsong will discuss how to more easily create super resolution images.
Digital Image Processing: Point and Local Operation Filters

Digital Image Processing: Point and Local Operation Filters

Experts
Heiko Gäthje
Senior Trainer, Training Academy
Olympus Europa
Takeo Ogama
Senior Product and Strategy Planner and Product Manager
Images captured with a light microscope are never true representations of the specimen; there are always sources of error that must be controlled. In this webinar, we will discuss how these sources of error can be managed.
Think Deep, See Deeper with Near-Infrared Laser Scanning Confocal Microscope

Think Deep, See Deeper with Near-Infrared Laser Scanning Confocal Microscope

Experts
Chunsong Yan
Business Development Manager, Life Science
Olympus Australia & New Zealand
Srivats Hariharan
Manager, Product and Application at Olympus Singapore
If you’re interested in having more fluorescent markers in your sample, imaging deeper into your sample at high resolution, and conducting live cell imaging with minimal phototoxicity, check out this on-demand webinar from Olympus imaging experts.
Light Sheet Microscopy for Deeper Insight into Life

Light Sheet Microscopy for Deeper Insight into Life

Experts
Irina Rakotoson
Life Science Product Manager, PhaseView
In this webinar, you’ll learn how the Alpha3 light sheet microscope combines very thin optical sectioning and high-quality Olympus optics for high-resolution 3D imaging of both live and fixed biological samples.
Imaging Multiple Subcellular Structures at the Nanometer Scale in 3D

Imaging Multiple Subcellular Structures at the Nanometer Scale in 3D

Experts
Dr. Nicolas Bourg
Chief Technical Officer and Co-Founder of Abbelight
Join Dr. Nicolas Bourg, the co-founder of Abbelight, as he discusses how fast and easy it can be to obtain high-quality, reliable, and understandable single-molecule images of your samples with 15 nm resolution in 3D.
FluidFM: A Novel Approach to CRISPR Gene Editing by Direct Intra-Nuclear Delivery

FluidFM: A Novel Approach to CRISPR Gene Editing by Direct Intra-Nuclear Delivery

Experts
Paul Monnier
Life Science Application Scientist
In this webinar, Paul Monnier from Cytosurge will show you how to overcome one of the biggest challenges in gene editing: delivery of the genetic material into the nucleus. Paul will discuss the FluidFM nanosyringe that can gently pierce the nuclear envelope to deliver reagents directly into the nucleus.
3D Assays: Intelligent Software, Insightful Analyses

3D Assays: Intelligent Software, Insightful Analyses

Experts
Daniel Bemmerl
Area Manager, High-End Microscopy Systems
Shohei Imamura
Strategic Project Manager, Regenerative Medicine and Drug Discovery Solutions
In this webinar, we discuss how to obtain the best statistical data for spheroids and microplate-based assays. These data can help you quantify the response of a 3D model to a compound and makes it easier to compare the effect of different treatments, such as varying concentrations.
Multiplexing and Deep Tissue Imaging with Near-Infrared Confocal Laser Scanning Microscopy

Multiplexing and Deep Tissue Imaging with Near-Infrared Confocal Laser Scanning Microscopy

Experts
Bülent Peker
Senior Product Marketing Manager
Olympus Europa
Rebecca Bonfig
Product Manager, Confocal Microscopy
In this webinar, you’ll learn about fluorescence multiplexing and deep tissue imaging using near-infrared (NIR) laser light.
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