Hokkaido University Faculty of Medicine, Physiological Science, Physiology
Could you please tell us about your scientific background and research interest?
Prof. Ohba, Hokkaido University
I obtained my PhD from the University of Tokyo as a cell biologist. I did my post-doc here at Hokkaido University in the laboratory of Prof. Yusuke Ohba, where I gained a lot of experience in microscopy, especially in live cell imaging. We have an interest in the spatial temporal relation of signal transduction including membrane trafficking and organelle biology. In particular, I’m studying the mechanism by which viruses enter host cells and the cellular responses to the viruses.
When you had to buy a new microscope, how did you end up choosing CrestOptics systems?
We used another spinning disk microscope, but the field of view was not wide enough, so we looked for a confocal spinning disk with a larger field of view. Also, we previously purchased the X-Light V2 which is a very good system and so we easily choose the X-Light V3 spinning disk.
Can you please describe your microscope set-up?
In the laboratory of Prof. Ohba, we have the X-Light V3 spinning disk combined with DeepSIM X-Light on an inverted Nikon Ti with stage top incubator, dual Prime BSI cameras and operated with VisiView software.
Which features of X-Light V3 spinning disk are helping your research?
We previously had a spinning disk microscope which we used to observe many samples, but we were not able to image many cells at once. Instead, with the X-Light V3, we could see many more cells in a single field of view thanks to its larger size of 25 mm. Moreover, we usually use an endosomal marker protein and overexpression of such protein causes mislocalization to the cytoplasm, although fluorescence is easily detected. But when the expression is low, we cannot see it using conventional confocal microscopy. Instead, the X-Light V3 allows us to easily detect even such low-intensity signals, and we are extremely happy about this. This happened in many other organelles and thanks to the X-Light V3 high throughput we could observe this event. We were also able to resolve very small organelles, so the resolution of the system is another characteristic that is of great importance.
"We are able to resolve very small organelles, so the resolution of the CrestOptics system is another characteristic of great importance for us”
What is your experience with the DeepSIM super-resolution system instead?
We have tried many super-resolution systems, but it was difficult to perform super-resolution imaging of our samples in living cells. For example, we could not detect weak signals. Nowadays, we can easily perform super-resolution imaging in a variety of living samples using the DeepSIM without stress in three different colours and using a stage top incubator.
“Nowadays, we can easily perform super-resolution imaging in a variety of living samples using the DeepSIM”
Are there other applications that utilize the CrestOptics setup in your lab?
We have many collaborators and perform imaging for them. A couple of examples are the imaging of brain tissues to study exosome transport, cell fusion to study the properties of SARS-CoV-2 variants, and virus entry to host cells to study HBV infection.
You invited us to participate at the Imaging bootcamp you’re organizing. Would you like to share some more insights about it?
We run the Imaging boot camp every year, this is going to be the 10th edition which will start on the 16th and finish on the 20th of January 2023. The topics are spinning disk confocal and super-resolution microscopy. It will be a mix of theory and practical sessions, with demonstrations and hands-on sessions using two CrestOptics configurations: the X-Light V3+DeepSIM X-Light and DeepSIM stand-alone system.
Thank you to Givetech for installation support.
