Step by Step Guide to Optimized Sample Preparation for GSD Microscopy

Authors: 1 , 1
Institute: 1Leica Microsystems, Wetzlar, Germany
13. January 2012

Ground state depletion microscopy followed by individual molecule return (GSDIM) is a super-resolution technique based on single molecule localization. To build a high-resolution image the ensemble of overlapping fluorophores in the diffraction-limited image has to be temporally “separated” to allow the high precision detection of single dyes.To achieve the optimal (super-)resolution the overall system, including the biological samples as well as optical, electronical and mechanical components, needs to be optimally integrated. Commercial solutions for GSD take care of the microscope components, and it is up to the researcher to prepare the sample in an optimal manner. This video tutorial presents a reliable way to produce samples for super-resolution GSD imaging with a special focus on the mounting step. Stable and flat mounting of the coverslip increases the performance of the overall system, leading to an improved resolution of the GSD image.

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Marko Lampe, Dr.

Marko Lampe joined Leica Microsystem in Wetzlar, Germany, in March 2011 as Application Manager for widefield super-resolution microscopy and cell biology applications. He studied Biochemistry at the University of Hanover and focused on single molecule detection fluorescently labeled motor-proteins with TIR-FM in vitro.

During his Ph.D. thesis he studied the entry of fluorescently labeled HIV-1 particles into the host cell at the University Hospital of Heidelberg, Germany. From 2008 until 2011, he worked as a postdoctoral fellow at the MRC Laboratory of Molecular Biology (LMB) in Cambridge, UK, studying clathrin-mediated endocytosis of GPCRs with live cell fluorescence microscopy.

Marko.Lampe@leica-microsystems.com

Beate Braun

Beate Braun is working for the "Application Management Widefield" team of Leica Microsystems in Wetzlar. Her main responsibilities are the organization of launch/training events for "Widefield Marketing" and the creation of tutorial videos about special applications.

After completing a professional education as Industral Sales Management Assistant, she studied English, Business Administration and Spanish at the Justus Liebig University of Gießen, Germany, focussing on linguistics and international marketing. She then worked for a communications agency for several years, before joining the Marketing department of Leica Microsystems in 2001.

Beate.Braun@leica-microsystems.com

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