Interview with Mr. Michael Moore

Mr. Moore, could you please tell us about your scientific background and experience in the microscopy field?

Fresh out of college, I took a position at a local quality control laboratory for Playtex products (Dover, DE) where my primary responsibility was to measure the pH of sunscreen lotions.  Ultimately, I realized this was not what I really wanted to do. I applied for a position as a Departmental Assistant in the Biology department at Delaware State University (DSU), a Historically Black College University (HBCU).  I was hired to prepare basic biology and ecology labs for major and non-major undergraduates.  I worked in this capacity for 12 years.  During my time at DSU, in the summer months, I joined a microscopy laboratory, researching single molecule and fluorescence imaging.  The summers I did research in that lab piqued my interest in microscopy. I began my career in microscopy at the Bioimaging facility at the Delaware Biotechnology Institute (DBI), the University of Delaware in 2012.  I was in charge of confocal microscopy training and supervised usage.  After 5 years I left the DBI Bioimaging core in 2017 to pursue a career as the OSCAR Imaging Facility Manager. Since July 2023, I have taken on the role of Associate Director of the facility. 

X-Light V3 spinning-disk acquisition of a plant section.

Digitaria exilis (Fonio) imaging to follow chlorophyll degradation or amplification after specific treatments. 10X objective, montage with Z stack.

What is your current role in the OSCAR facility and your scientific area of interest?

I am currently the Associate Director after serving as the OSCAR Imaging Facility Manager for 5 years. My primary area of interest is running a core facility, and assisting users and researchers to collect the best possible data. It is my responsibility to train and supervise students and researchers on our various instruments. I am also in charge of all microscope maintenance and repairs.

In addition, my research interest is understanding the latest technology available in microscopy and spectroscopy. I am personally interested in using fluorescence (FCS), SHG imaging, and spectroscopy to characterize the movement of molecules across cellular systems and membranes, as well as the use of novel imaging/spectroscopy techniques in the characterization of microplastics from environmental and biological samples.

In the OSCAR imaging facility, we are bringing together known imaging approaches (e.g., Widefield, Confocal) with new modalities, including NanoIR AFM, and Raman confocal.  We are also interested in spinning disks and super-resolution microscopy; therefore, we are installing an X-Light V3 spinning disk and a DeepSIM stand-alone from CrestOptics.

Can you tell us what instruments are available to the scientific community at OSCAR? 

Our facility serves the entire research community at the DSU. This includes over 40 diverse laboratory groups, including a diverse group of graduate and undergraduate students. We also serve 7 industry partners and external users. We offer confocal, super-resolution, electron, and atomic force microscopy, Raman confocal microscopy, and a host of spectroscopy-based instruments.

"The main advantages of the CrestOptics system are speed, sensitivity, ease of use, and flexibility”

Mr. Moore and PhD student Tiffany Bamfo (Harrington Lab, Delaware State University) are using the X-Light V3 spinning disk to perform Calcium imaging in Spinal Muscular Atrophy (SMA) cells.

OSCAR offers a wide range of high-end imaging systems. What are the main advantages of CrestOptics systems, in your opinion? 

The X-Light V3 and DeepSIM systems can be used to fill practically all the confocal and super-resolution applications of our user base. The ability to upgrade and swap out cameras is a definite plus and does not lock the instrument into a specific camera manufacturer or type. The flexibility of having customized disk boxes for specific applications, and the ability to change out disk patterns due to the sealed disk box design are notable advantages not available on competing systems. CrestOptics products are very cost-effective ways to collect and evaluate intricate biological processes. Moreover, the capability to mount on almost any microscope stand, including upright systems, makes these instruments very flexible. Finally, the CrestOptics systems can be used with several pieces of software, and this is important for flexibility in an imaging facility

Live answer

"Such a description of the setup makes it clear that flexibility and compatibility are the driving factors in making our decision to purchase a CrestOptics Spinning Disk Confocal”

What are your CrestOptics X-Light V3 imaging set-ups like? Which features of the X-Light V3 spinning disk are most helpful to your research? 

We have an X-Light V3 mounted to the side port of a IX83 body (Olympus). We are currently using two ORCA Fusion 2 cameras (Hamamatsu), a 6-line LDI laser system (89 North), and Ludl stage with a piezo insert. Moreover, the system is equipped with RapOptics and Visitron controller for FRAP with a 405 Laser. All devices are driven by Visiview software. 

The current setup we have allows us to take fast montage / z- stack projects that would usually take hours on our point scanning confocal down to just minutes. The ability to capture calcium signals in real-time has helped in pinpointing cellular events we could not have resolved using our point scanning confocal. The time-saving aspects of the spinning disk have increased productivity. 

Microscope set-ups are located at Optical Science Center for Applied Research  / OSCAR (Delaware State University). Thanks to our distributor Voxyl Microscopes.

For what applications is a DeepSIM super-resolution system mainly used and what are the advantages? 

The DeepSIM will be used for many of the projects that require that bump in the resolution that we normally would farm out to the DBI Bioimaging core. We currently have several projects that require resolutions approaching 100 nm and the DeepSIM will be used for these projects.  For instance, the DeepSIM is used for Caenorhabditis elegans studies, neuronal/astrocyte synaptic interactions, and brain co-localization in Parkinson’s disease mouse brain sections.

Live answer

Overall, what are all the biological applications that you are carrying out on the CrestOptics systems?

The current biological application we are using our CrestOptics systems for is C. elegans neuronal distribution experiments, Drosophila whole brain imaging, calcium imaging in spinal muscular atrophy (SMA), astrocyte and neuron interactions in mixed cultures. Moreover, whole mouse brain montages are helpful for neuronal disease research such as Parkinson’s, whereas organelle tracking is relevant in Alzheimer’s disease mouse cellular models.   

In my opinion, The X-Light V3 and DeepSIM stand-alone systems make the most sense in a core facility. The performance of both the stand-alone systems is superb and feedback from users is very positive. The stand-alone systems allow one user to use the DeepSIM while a second user can use the X-Light V3 without interrupting each other. 

X-Light V3 spinning-disk dual-camera live cell imaging.

Chicken astrocytes infected with the Calcium sensor GCamp6F (Red channel). The addition of Glutamate induces the elevation of calcium (Green channel). Imaged at 15 FPS using a 20X 0.8 N.A. Oil, UPLanSApo. Courtesy: Temburni Lab Delaware State University

In your opinion, what is the future of fluorescence microscopy and imaging?

We imagine the ability to use machine learning and machine vision will help to locate and analyze samples to better aid the researcher and technicians in core facilities. Coupling the ability to auto-find regions of interest in cells or samples can be integrated into fluorescence systems due to the discrete signals of fluorophores. The addition of spectroscopic analysis coupled into a super-resolution system would add contextual details in a correlative capacity.  

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