Abstract
Super-resolution microscopy includes multiple techniques in optical microscopy that enable sub-diffraction resolution fluorescence imaging of cellular structures. Expansion microscopy (EXM) is a method of physical expansion to obtain super-resolution images of a biological sample on conventional microscopy. We present images of yeast organelles, applying the combination of super-resolution and ExM techniques. When preparing pre-expanded samples, conventional methods lead to breakage of dividing yeast cells and difficulties in studying division-related proteins. Here, we describe an improved sample preparation technique that avoids such damage. ExM in combination with Airyscan and structured illumination microscopy (SIM) collected sub-cellular structural images of nuclear pore complex, septin, and a-tubulin in yeast. Our method of expansion in yeast is well-suited for super-resolution imaging study of yeast.
Highlights
Optical microscopy is one of the most important tools in the fields of biology and medicine
The traditional immunofluorescence approach may result in structures such as septin being severely damaged during the preparation of spheroplasts and centrifugation
We illuminated the nuclear pore complex (NPC) located on the equator of the yeast nucleus, which display a circular distribution of nuclear pore spots (Figure 1B)
Summary
Optical microscopy is one of the most important tools in the fields of biology and medicine. Different methods have emerged to achieve optical super-resolution imaging, breaking the limits of diffraction. These super-resolution microscopy techniques can be divided into two categories—light source modulation and singlemolecule modulation. Airyscan microscopy is a new detector design that optimizes classic components, consisting of 32 elements acting as its own small pinhole in an Airyscan with positional information. This design increases the contrast of the high spatial frequency information but extends the imaging time [8]. Each of the aforementioned super-resolution techniques have its own advantages and disadvantages, which are applicable to different types of samples for increased resolution
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
