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  • Bridging Structure and Dynamics at the Nanoscale through Optogenetics and Electrical Stimulation

    Nanoscale ultrastructural information is typically obtained by means of static imaging of a fixed and processed specimen. However, this is only a snapshot of one moment within a dynamic system in…
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  • Superior Ultrastructural Preservation and Structural Contrast in Drosophila Tissue

    Optimal structural preservation of tissue can only be obtained by high pressure freezing. However, preparing the samples in optimal conditions is challenging. This article explains in detail how to…
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  • Maple (Acer saccharum) Leaves - High Pressure Freezing and Freeze Substitution for TEM

    Application Note for Leica EM HPM100 - Leaves were immersed in hexadecene and placed under a gentle (0.3 bar) vacuum for 10 minutes to evacuate the internal air spaces. The leaves were then trimmed to…
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  • High-Pressure Freezing and Freeze Substitution of Hep-2 Cells Infected with Chlamydia pneumoniae

    Application Note for Leica EM HPM100 - Hep-2 cells infected with Chlamydia pneumoniae were cultured on carbon-coated 6 mm Sapphire discs. Cells were high-pressure frozen in an EM HPM100 using the 6 mm…
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  • Visualization of Membrane Dynamics with Millisecond Temporal Resolution

    Application Note for Leica EM ICE, Leica EM AFS2 - Electrical stimulation of neurons combined with high-pressure freezing allows physiological activation of synaptic activity and precise control over…
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  • Structural Study of C. elegans

    Application Note for Leica EM ICE, Leica EM AFS2 - Wildtype L4 stage C. elegans (N2 strain) were placed in the 100 μm deep side of Lecithin-coated (see detailed protocol*) type A 3 mm Cu/Au carriers…
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  • Cultured Rat Hippocampal Neurons

    Application Note for Leica EM ICE - Rat Hippocampal neurons, cultured on 50 μm thick Aclar (Aclar embedding film, EMS) for 19 days, were frozen in the 100 μm deep side of lecithin coated (detailed…
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  • Immuno - Electron Microscopy of High Pressure Frozen and Freeze Substituted Mouse Heart

    Application Note for Leica EM AFS2 - Mouse heart tissue from wild-type (WT) and αT -catenin KO (KO) mice was excised, immersed in 20% (w/v) BSA and frozen immediately in a high-pressure freezer Leica…
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  • Cell Envelope Structure of a Gram-negative Thermotolerant y-Proteobacterium Acidithiobacillus sp., Strain HV2/2 and its Interaction with Pyrite

    Application Note for Leica EM AFS2 - Cells of the the moderately thermophilic Acidithiobacillus sp.strain HV2/2, were centrifuged at 20,0009xg and then cryo-immobilized by high-pressure freezing on…
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  • Symmetric Synapse - Clathrin Coated Endocytosis Pit in the Postsynaptic Dendrite

    Application Note for Leica EM ICE - WT hippocampal neurons were plated at a density of 80,000 cells/cm2 on 6 mm sapphire disks for 14 days. Sample were frozen using a high-pressure freezer (Leica EM…
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  • Electron Microscopy of High Pressure Frozen and Freeze Substituted Arabidopsis Thaliana Root Tips Cells

    Application Note for Leica EM AFS2 - Arabidopsis thaliana roots (mutant PIN1pro:PIN1-GFP;bex5-1) were excised, immersed in 20% (w/v) BSA and frozen immediately in a high-pressure freezer. Freeze…
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  • Transmission Electron Microscopy Sample Preparation Protocols for the Ultrastructural Study of Cysts of Free-living Protozoa

    Cysts of free-living protozoa have an impact on the ecology and epidemiology of bacteria because they may act as a transmission vector or shelter the bacteria against hash environmental conditions.…
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  • Correlative In-Resin Super-Resolution and Electron Microscopy Using Standard Fluorescent Proteins

    We introduce a method for correlative in-resin super-resolution fluorescence and electron microscopy (EM) of biological structures in mammalian culture cells. Cryo-fixed resin embedded samples offer…
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  • New Insights into Cilia and Flagella by Cryo-EM

    Cilia and flagella were the first organelles to be discovered and have been studied for centuries. However, their essential role in humans and how ciliary defects cause diseases are still not well…
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  • Electron Tomography of Cryo-Immobilized Plant Tissue: A Novel Approach to Studying 3D Macromolecular Architecture of Mature Plant Cell Walls In Situ

    Cost-effective production of lignocellulosic biofuel requires efficient breakdown of cell walls present in plant biomass to retrieve the wall polysaccharides for fermentation. In-depth knowledge of…
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  • Capturing Cellular Dynamics with Millisecond Temporal Resolution

    The combination of two powerful techniques: optogenetics and high-pressure freezing now makes it possible to visualize a dynamic cellular activity with temporal resolution of 5 milliseconds. By…
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  • Freeze Substitution of Trypanosoma brucei

    Chemical fixation of biological specimens for ultrastructural investigation is a relatively slow and selective process, and therefore a common source of artifacts. Freezing, on the other hand, is an…
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  • Brief Introduction to Freeze Substitution

    Freeze-substitution is a process of dehydration, performed at temperatures low enough to avoid the formation of ice crystals and to circumvent the damaging effects observed after ambient-temperature…
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  • From Dynamic Live Cell Imaging to 3D Ultrastructure: Novel Integrated Methods for High Pressure Freezing and Correlative Light-Electron Microscopy

    To correlate dynamic events in adherent cells with both ultrastructural and 3D information, we developed a method for cultured cells that combines confocal time-lapse images of GFP-tagged proteins…
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  • Targeting of Peroxisomal Matrix Proteins in the Diatom Phaeodactylum Tricornutum

    P. tricornutum cells expressing different types of GFP fusion proteins were harvested via centrifugation at 1,500xg and cryoimmobilized by high-pressure freezing on gold carriers.
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