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Imaging Multiple Subcellular Structures at the Nanometer Scale in 3D

During this webinar, Nicolas will show you 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. He will go through the entire imaging workflow—from sample preparation to data analysis—to show you what the single-molecule local microscopy (SMLM) community has made possible, what the common mistakes are, and how Abbelight’s tools and support can make the imaging and analysis journey much easier.

Presenter:

Dr. Nicolas Bourg, Chief Technical Officer and Co-Founder of Abbelight

FAQ

Webinar FAQs | Imaging multiple subcellular structures at the nanometer scale

How long does it take to acquire 3D multicolor nanoscopy images?

The spectral demixing strategy means that acquisition of three-color images typically takes less than one minute for a standard field of view in 3D (about 50 × 50 µm), and a few minutes for larger fields of view (150 × 150 µm).

How difficult is it to prepare nanoscopy structures?

Sample preparation requires only simple modification of the standard preparation procedure, such as changing the concentration of the fixative (e.g., PFA) or increasing the antibody concentration. It only requires fine tuning of your own protocol. The Abbelight team can provide guidance for this if necessary.  

How do you deal with chromatic aberrations in multi-color experiments and with sample drifts?

The spectral demixing strategy allows simultaneous imaging of 2 to 3 far-red channels like 647 nm, 660 nm, and 680 nm. Since the 3 colors are very close in wavelength, there is no chromatic aberrations with this technique.

Sample drifts are always calculated and applied on the image in real time during the acquisition, and they can be refined with different published algorithms and methods in post-acquisition processing.

What is the importance of nanoscopy in diagnosing diseases such as Parkinson’s, Alzheimer’s, and viral infection?

The power of SMLM nanoscopy relies on the fact that the output of an acquisition is not only an image, but a text file with the X,Y, and Z position of every molecule detected during the acquisition. This enables:

  • Quantification of the targeted molecules (up to 3 simultaneously), for instance to follow the evolution of clusters of molecules under drug therapy
  • Nanometric 3D spatial information for structural analysis or colocalization of proteins
  • Tracking of individual molecules in time for dynamic analysis

This potential is currently used to help diagnose diseases, develop novel therapies, diagnose viral or bacterial infection, and much more.

Imaging Multiple Subcellular Structures at the Nanometer Scale in 3D

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Ask the Experts: Imaging multiple subcellular structures at the nanometer scale in 3D

During this webinar, Nicolas will show you 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. He will go through the entire imaging workflow—from sample preparation to data analysis—to show you what the single-molecule local microscopy (SMLM) community has made possible, what the common mistakes are, and how Abbelight’s tools and support can make the imaging and analysis journey much easier.

FAQ

Webinar FAQs | Imaging multiple subcellular structures at the nanometer scale

How long does it take to acquire 3D multicolor nanoscopy images?

The spectral demixing strategy means that acquisition of three-color images typically takes less than one minute for a standard field of view in 3D (about 50 × 50 µm), and a few minutes for larger fields of view (150 × 150 µm).

How difficult is it to prepare nanoscopy structures?

Sample preparation requires only simple modification of the standard preparation procedure, such as changing the concentration of the fixative (e.g., PFA) or increasing the antibody concentration. It only requires fine tuning of your own protocol. The Abbelight team can provide guidance for this if necessary.  

How do you deal with chromatic aberrations in multi-color experiments and with sample drifts?

The spectral demixing strategy allows simultaneous imaging of 2 to 3 far-red channels like 647 nm, 660 nm, and 680 nm. Since the 3 colors are very close in wavelength, there is no chromatic aberrations with this technique.

Sample drifts are always calculated and applied on the image in real time during the acquisition, and they can be refined with different published algorithms and methods in post-acquisition processing.

What is the importance of nanoscopy in diagnosing diseases such as Parkinson’s, Alzheimer’s, and viral infection?

The power of SMLM nanoscopy relies on the fact that the output of an acquisition is not only an image, but a text file with the X,Y, and Z position of every molecule detected during the acquisition. This enables:

  • Quantification of the targeted molecules (up to 3 simultaneously), for instance to follow the evolution of clusters of molecules under drug therapy
  • Nanometric 3D spatial information for structural analysis or colocalization of proteins
  • Tracking of individual molecules in time for dynamic analysis

This potential is currently used to help diagnose diseases, develop novel therapies, diagnose viral or bacterial infection, and much more.

Experts
Nicolas Bourg博士
Abbelight首席技术官兼联合创始人

大家好。我是Abbelight的首席技术官(CTO)兼联合创始人Nicolas Bourg博士,是您的专职单分子定位显微镜(SMLM)专家- 这类显微镜也被称为纳米显微镜-其中包括PALM、dSTORM、SPT-PALM和DNA-PAINT等技术。

我是一名经过培训的光电工程师,并在巴黎-萨克雷大学获得生物光子学研究领域的博士学位,致力于通过独特3D纳米技术实现前所未有的高分辨率的研究。我与我的研究团队创建了Abbelight这家初创公司,决定分享我在博士学位期间获得的所有知识,致力于让纳米显微镜功能更加强大,且让任何生物学家无需进行高级培训即可掌握显微镜的全面操作。我的工作任务是回答您有关纳米显微镜的所有问题,欢迎与我联系。

Imaging Multiple Subcellular Structures at the Nanometer Scale in 3D2021年12月3日
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