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6. Cost-efficient nanoscopy reveals nanoscale architecture of liver cells and platelets

Author

Mao Hong, Diekmann Robin, Liang Hai Po H., Cogger Victoria C., Le Couteur David G., Lockwood Glen P., Hunt Nicholas J., Schüttpelz Mark, Huser Thomas R., Chen Vivien M., and McCourt Peter A.G.

Subjects
liver, endothelium, optical nanoscopy, fenestration, platelet, Physics, QC1-999
Abstract

Single-molecule localization microscopy (SMLM) provides a powerful toolkit to specifically resolve intracellular structures on the nanometer scale, even approaching resolution classically reserved for electron microscopy (EM). Although instruments for SMLM are technically simple to implement, researchers tend to stick to commercial microscopes for SMLM implementations. Here we report the construction and use of a “custom-built” multi-color channel SMLM system to study liver sinusoidal endothelial cells (LSECs) and platelets, which costs significantly less than a commercial system. This microscope allows the introduction of highly affordable and low-maintenance SMLM hardware and methods to laboratories that, for example, lack access to core facilities housing high-end commercial microscopes for SMLM and EM. Using our custom-built microscope and freely available software from image acquisition to analysis, we image LSECs and platelets with lateral resolution down to about 50 nm. Furthermore, we use this microscope to examine the effect of drugs and toxins on cellular morphology.

Published
2019
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9. Effect of caffeine and other xanthines on liver sinusoidal endothelial cell ultrastructure

Author

Mao, Hong, primary, Szafranska, Karolina, additional, Kruse, Larissa, additional, Holte, Christopher, additional, Wolfson, Deanna L., additional, Ahluwalia, Balpreet Singh, additional, Whitchurch, Cynthia B., additional, Cole, Louise, additional, Lockwood, Glen P., additional, Diekmann, Robin, additional, Le Couteur, David, additional, Cogger, Victoria C., additional, and McCourt, Peter A.G., additional

Published
2023
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12. Implementation of a 4Pi-SMS super-resolution microscope

Author

Wang, Jingyu, primary, Allgeyer, Edward S., additional, Sirinakis, George, additional, Zhang, Yongdeng, additional, Hu, Kevin, additional, Lessard, Mark D., additional, Li, Yiming, additional, Diekmann, Robin, additional, Phillips, Michael A., additional, Dobbie, Ian M., additional, Ries, Jonas, additional, Booth, Martin J., additional, and Bewersdorf, Joerg, additional

Published
2020
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15. Organotypic slice culture model demonstrates interneuronal spreading of alpha-synuclein aggregates

Author

Elfarrash, Sara, primary, Jensen, Nanna Møller, additional, Ferreira, Nelson, additional, Betzer, Cristine, additional, Thevathasan, Jervis Vermal, additional, Diekmann, Robin, additional, Adel, Mohamed, additional, Omar, Nisreen Mansour, additional, Boraie, Mohamed Z., additional, Gad, Sabry, additional, Ries, Jonas, additional, Kirik, Deniz, additional, Nabavi, Sadegh, additional, and Jensen, Poul Henning, additional

Published
2019
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16. Characterization of an industry-grade CMOS camera well suited for single molecule localization microscopy – high performance super-resolution at low cost

Author

Diekmann, Robin, Till, Katharina, Müller, Marcel, Simonis, Matthias, Schüttpelz, Mark, and Huser, Thomas

Subjects
lcsh:R, ComputingMethodologies_IMAGEPROCESSINGANDCOMPUTERVISION, lcsh:Medicine, lcsh:Q, lcsh:Science, Article
Abstract

Many commercial as well as custom-built fluorescence microscopes use scientific-grade cameras that represent a substantial share of the instrument’s cost. This holds particularly true for super-resolution localization microscopy where high demands are placed especially on the detector with respect to sensitivity, noise, and also image acquisition speed. Here, we present and carefully characterize an industry-grade CMOS camera as a cost-efficient alternative to commonly used scientific cameras. Direct experimental comparison of these two detector types shows widely similar performance for imaging by single molecule localization microscopy (SMLM). Furthermore, high image acquisition speeds are demonstrated for the CMOS detector by ultra-fast SMLM imaging.

Published
2017

19. Chip-based wide field-of-view nanoscopy

Author

Diekmann, Robin, primary, Helle, Øystein I., additional, Øie, Cristina I., additional, McCourt, Peter, additional, Huser, Thomas R., additional, Schüttpelz, Mark, additional, and Ahluwalia, Balpreet S., additional

Published
2017
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21. Nanoscopy of bacterial cells immobilized by holographic optical tweezers

Author

Diekmann, Robin, Wolfson, Deanna L., Spahn, Christoph Klaus, Heilemann, Mike, Schüttpelz, Mark, Huser, Thomas, Diekmann, Robin, Wolfson, Deanna L., Spahn, Christoph Klaus, Heilemann, Mike, Schüttpelz, Mark, and Huser, Thomas

Abstract

Imaging non-adherent cells by super-resolution far-field fluorescence microscopy is currently not possible because of their rapid movement while in suspension. Holographic optical tweezers (HOTs) enable the ability to freely control the number and position of optical traps, thus facilitating the unrestricted manipulation of cells in a volume around the focal plane. Here we show that immobilizing non-adherent cells by optical tweezers is sufficient to achieve optical resolution well below the diffraction limit using localization microscopy. Individual cells can be oriented arbitrarily but preferably either horizontally or vertically relative to the microscope’s image plane, enabling access to sample sections that are impossible to achieve with conventional sample preparation and immobilization. This opens up new opportunities to super-resolve the nanoscale organization of chromosomal DNA in individual bacterial cells.

Published
2016

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