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

Live cell super resolution imaging: The big picture of small things

In this webinar, Stefan, Lauren, and Chunsong will discuss how to more easily create super resolution images. Olympus super resolution uses the IXplore SpinSR spinning disc confocal system to combine speed, sensitivity, and resolution for live-cell-compatible super resolution imaging. They will explain the steps involved to create super resolution images, including optical reassignment, Fourier analysis, and 3D deconvolution.

Presenters:

Stefan Marawske, Senior Sales Specialist, High-End Life Science Systems
Lauren Alvarenga, Product Manager, Life Science Microscopy
Chunsong Yan, Business Development Manager, Life Science

FAQ

Webinar FAQs | Live Cell Super-Resolution Imaging

What are the advantages of the IXplore SpinSR system over widefield-based structured illumination microscopes?

Speed is one of the major advantages of working with the IXplore SpinSR microscope. On widefield-based microscopes, the grid has to rotate many times, and the camera’s exposure time limits speed on the multiple exposures. On the IXplore SpinSR system, the spinning-disk-based optics eliminate the need for a structured illumination grid so that you can collect images faster and with fewer frames. Another advantage is that the IXplore SpinSR microscope has better depth capability—you can use a thicker specimen and still get a better image because of its confocal capabilities.

Is 3D deconvolution necessary to achieve 120 nm resolution?

The dedicated hardware optics and the Olympus Super Resolution (OSR) algorithm are the system’s key components that achieve a 120 nm resolution. However, deconvolution will further enhance the image quality and contrast.

What software runs the Olympus IXplore SpinSR microscope?

The system is run by Olympus cellSens software, which is widely used for Olympus microscopes and can instantly recognize the correct hardware. Moreover, the user interface is straightforward and intuitive. 

What kinds of detection systems are available with the Olympus IXplore SpinR microscope, and what’s the frame rate?

When imaging, the smaller the pixel, the better the resolution. With smaller pixels, you need to have better sensitivity and you also need to be careful of noise, so a Peltier-cooled scientific CMOS camera is the preferred option. We use a 2k × 2k scientific camera with a quantum efficiency of more than 80%. However, the main limitation is the sample. If you have a brighter sample, your camera’s exposure time decides the frame rate, and you won’t have to acquire multiple images to construct one super resolution image. So, if for every exposure you get one super-resolved image, you reach up to 100 frames per second. In addition, the system gives you the option to have two cameras, enabling you to image two colors simultaneously. However, the system also comes with a built-in fast emission filter that enables you to do fast sequential multicolor imaging, depending on the number of lasers and emission filters.

What types of lasers does the IXplore SpinSR system come with?

We provide all the usual lasers that conventional confocal microscopes have. We have solid state lasers that are very robust and stable and last longer than conventional gas lasers, with 405, 488, 560, and 640 as standard.

Can you use the Olympus IXplore SpinSR system for high-content screening?

Yes, absolutely. Olympus offers the advanced scanR high-content screening software. It comes with many unique features, such as the latest self-learning AI module (deep learning). It has a powerful interactive interface, can perform multilevel scanning, and is extremely versatile. Specifically concerning high-content screening, it offers exceptional speed and accuracy due to the parallel acquisition feature built into the scanR software.

Is it possible to carry out photo stimulation experiments on the IXplore SpinSR system in combination with confocal imaging?

Yes. Also, the system’s real-time controller can control older hardware through the TTL, which is much more accurate for time-dependent experiments.

Do your objective lenses have correction collars for spherical aberrations? Are they easy to adjust?

We offer A Line application-driven objectives with correction collars and a silicone immersion objective that has a long working distance. It has a correction collar, and the silicone oil more closely matches the refractive index of the sample, so you can image deeper with less aberrations. We also have some special high numerical aperture objectives, plan apochromat lenses that have an NA of 1.5 and come with a correction collar that you can set at different temperatures. So, ideally you can go up to 200 microns if you’re using a 30x silicone immersion objective.


相关产品

超分辨率显微镜系统

IXplore SpinSR

  • 超级分辨率可让XY分辨率达到120 nm
  • 更低的光毒性和光漂白可以在共聚焦时间序列成像时降低对细胞活力的影响
  • 宽场、共聚焦和超分辨模式一键式切换
  • 使用世界首个数值孔径(NA)值1.5的平场复消色差物镜进行精确的3D重构*
    *截至2018年11月。根据奥林巴斯的研究。

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

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

In this webinar, Stefan, Lauren, and Chunsong will discuss how to more easily create super resolution images. Olympus super resolution uses the IXplore SpinSR spinning disc confocal system to combine speed, sensitivity, and resolution for live-cell-compatible super resolution imaging. They will explain the steps involved to create super resolution images, including optical reassignment, Fourier analysis, and 3D deconvolution.

FAQ

Webinar FAQs | Live Cell Super-Resolution Imaging

What are the advantages of the IXplore SpinSR system over widefield-based structured illumination microscopes?

Speed is one of the major advantages of working with the IXplore SpinSR microscope. On widefield-based microscopes, the grid has to rotate many times, and the camera’s exposure time limits speed on the multiple exposures. On the IXplore SpinSR system, the spinning-disk-based optics eliminate the need for a structured illumination grid so that you can collect images faster and with fewer frames. Another advantage is that the IXplore SpinSR microscope has better depth capability—you can use a thicker specimen and still get a better image because of its confocal capabilities.

Is 3D deconvolution necessary to achieve 120 nm resolution?

The dedicated hardware optics and the Olympus Super Resolution (OSR) algorithm are the system’s key components that achieve a 120 nm resolution. However, deconvolution will further enhance the image quality and contrast.

What software runs the Olympus IXplore SpinSR microscope?

The system is run by Olympus cellSens software, which is widely used for Olympus microscopes and can instantly recognize the correct hardware. Moreover, the user interface is straightforward and intuitive. 

What kinds of detection systems are available with the Olympus IXplore SpinR microscope, and what’s the frame rate?

When imaging, the smaller the pixel, the better the resolution. With smaller pixels, you need to have better sensitivity and you also need to be careful of noise, so a Peltier-cooled scientific CMOS camera is the preferred option. We use a 2k × 2k scientific camera with a quantum efficiency of more than 80%. However, the main limitation is the sample. If you have a brighter sample, your camera’s exposure time decides the frame rate, and you won’t have to acquire multiple images to construct one super resolution image. So, if for every exposure you get one super-resolved image, you reach up to 100 frames per second. In addition, the system gives you the option to have two cameras, enabling you to image two colors simultaneously. However, the system also comes with a built-in fast emission filter that enables you to do fast sequential multicolor imaging, depending on the number of lasers and emission filters.

What types of lasers does the IXplore SpinSR system come with?

We provide all the usual lasers that conventional confocal microscopes have. We have solid state lasers that are very robust and stable and last longer than conventional gas lasers, with 405, 488, 560, and 640 as standard.

Can you use the Olympus IXplore SpinSR system for high-content screening?

Yes, absolutely. Olympus offers the advanced scanR high-content screening software. It comes with many unique features, such as the latest self-learning AI module (deep learning). It has a powerful interactive interface, can perform multilevel scanning, and is extremely versatile. Specifically concerning high-content screening, it offers exceptional speed and accuracy due to the parallel acquisition feature built into the scanR software.

Is it possible to carry out photo stimulation experiments on the IXplore SpinSR system in combination with confocal imaging?

Yes. Also, the system’s real-time controller can control older hardware through the TTL, which is much more accurate for time-dependent experiments.

Do your objective lenses have correction collars for spherical aberrations? Are they easy to adjust?

We offer A Line application-driven objectives with correction collars and a silicone immersion objective that has a long working distance. It has a correction collar, and the silicone oil more closely matches the refractive index of the sample, so you can image deeper with less aberrations. We also have some special high numerical aperture objectives, plan apochromat lenses that have an NA of 1.5 and come with a correction collar that you can set at different temperatures. So, ideally you can go up to 200 microns if you’re using a 30x silicone immersion objective.


相关产品

超分辨率显微镜系统

IXplore SpinSR

  • 超级分辨率可让XY分辨率达到120 nm
  • 更低的光毒性和光漂白可以在共聚焦时间序列成像时降低对细胞活力的影响
  • 宽场、共聚焦和超分辨模式一键式切换
  • 使用世界首个数值孔径(NA)值1.5的平场复消色差物镜进行精确的3D重构*
    *截至2018年11月。根据奥林巴斯的研究。
Experts
Chunsong Yan
Process Systems业务发展经理
Olympus Australia & New Zealand

您好,我叫Chunsong Yan,我是奥林巴斯澳大利亚和新西兰公司生命科学业务发展经理。我目前负责共聚焦、多光子、光片和全玻片扫描系统。我于2003年加入奥林巴斯并任过各类职务,任职期间始终致力于为我们的客户提供最佳奥林巴斯解决方案。

Lauren Alvarenga
生命科学显微镜产品经理

大家好。我是Lauren Alvarenga,在奥林巴斯担任研究成像产品经理,目前负责成像软件以及倒置和超分辨率显微镜。我拥有罗彻斯特理工学院的生物医学摄影通讯专业理学学士学位。
甲状旁腺激素对肾脏磷酸转运蛋白Npt2a的转录后调节。我自2015年起一直在奥林巴斯工作,主要为美国、加拿大和拉丁美洲的FLUOVIEW系列产品提供支持。

Stefan Marawske
高端生命科学系统高级销售专员
Olympus Europa

您好,我叫Stefan Marawske,是您的超分辨率显微镜专家。在攻读物理化学领域博士学位期间,我曾制作过用于基于定位超分辨率和粒子跟踪的自搭建显微镜。我特别沉迷于那些能够突破著名的阿贝极限以及能够分辨以前无法观察结构的方法。

我在奥林巴斯工作了超过7年,主要负责TIRF和转盘等高端成像系统。这类系统通常由于能够将许多不同设备组合用于各种应用而具有高度的灵活性,因此,如果您在确定专用配置方面需要任何帮助,欢迎与我们联系。

Live Cell Super Resolution Imaging: The Big Picture of Small Things2021年12月3日
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