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Freeze Substitution

Freeze substitution is a technique used in electron microscopy to prepare biological samples for analysis.
It involves rapidly freezing the sample and then gradually replacing the frozen water with a solvent, such as acetone or methanol, at low temperatures.
This method preserves the ultrastructure of the sample and minimizes the artifacts that can occur with traditional chemical fixation.
Freeze substitution is particularly useful for studying the fine details of cellular organelles, membranes, and macromolecular complexes.
It is an important tool for researchers investigating the structure and function of biological systems at the nanoscale level.
The PubCompare.ai platform can help optimize freeze substitution protocols by providing access to relevant literature, preprints, and patents, as well as intelligent comparisons to identify the best methods and prodcuts for your research, streamlining your workflow and enusring reproducible results.

Most cited protocols related to «Freeze Substitution»

1
Cryo-Immobilization and Fluorescence Microscopy
Cited 14 times
S. pombe strain expressing RFP-mal3p and GFP-atb2p (gift of Stephen Huisman and Damian Brunner, EMBL, Heidelberg, Germany) and S.cerevisiae strains expressing EGFP- and mCherry-tagged endocytic proteins (described in Kaksonen et al., 2005 (link)) were grown in liquid YES medium and SD medium, respectively. All yeast cultures were grown at 30°C until reaching log phase, pelleted by filtering, and cryo-immobilized by HPF with a high pressure freezing system (EM PACT2; Leica).
MDCK-H2B-RFP cells (a gift of Daniela Holzer and Lars Hufnagel, EMBL, Heidelberg, Germany) were maintained in DME supplemented with 5% fetal calf serum and l-glutamine at 37°C under 5% CO2. Cells were grown on carbon-coated sapphire disks in 24-well dishes to 60–80% confluency. Purified HIV-eGFP-delEnv particles (Müller et al., 2004 (link); ∼250 ng p24/ well) provided by Manon Eckhardt (Universitätsklinikum Heidelberg, Heidelberg, Germany) were allowed to bind to the cell surface for 30–60 min on ice followed by cryo-immobilization by HPF with a high pressure freezing machine (HPM 010; BAL-TEC).
All samples were further processed by freeze substitution (FS) and embedding in a temperature-controlling device (model AFS2; Leica). FS occurred at −90°C for 48–58 h with 0.1% (wt/vol) uranyl acetate in glass-distilled acetone. Addition of 1–3% water, which is in some cases desired to improve membrane contrast (Walther and Ziegler, 2002 (link)), was tested and did not influence fluorescence retention. The temperature was then raised to −45°C (5°C/hour), and samples were washed with acetone and infiltrated with increasing concentrations (10, 25, 50, and 75%; 4 h each) of Lowicryl in acetone while the temperature was further raised to −25°C. 100% Lowicryl was exchanged three times in 10-h steps and samples were UV polymerized at −25°C for 48 h, after which the temperature was raised to 20°C (5°C/hour) and UV polymerization continued for 48 h. 300-nm sections were cut with a microtome (Ultracut UCT; Leica) and a diamond knife and picked up on carbon-coated 200 mesh copper grids. Blue (365 nm/415 nm) 0.02 µm FluoSpheres (Invitrogen) were pretreated (to reduce intensity) with 0.1% Tween 20 for 10 min, washed twice by ultracentrifugation at 100,000 g, resuspended in PBS, and adsorbed to the EM grids by placing the grids section face-down onto a 15-µl drop of FluoSpheres for 10 min. Grids were then washed with three drops of water and blotted with filter paper.
Kukulski W., Schorb M., Welsch S., Picco A., Kaksonen M, & Briggs J.A. (2011). Correlated fluorescence and 3D electron microscopy with high sensitivity and spatial precision. The Journal of Cell Biology, 192(1), 111-119.
Publication 2011
Acetone Carbon Cells Copper Diamond Face Fetal Bovine Serum Fluorescence Freeze Substitution Glutamine Hyperostosis, Diffuse Idiopathic Skeletal Immobilization Lanugo Madin Darby Canine Kidney Cells Medical Devices Microtomy Polymerization Pressure Proteins Retention (Psychology) Saccharomyces cerevisiae Sapphire Schizosaccharomyces pombe Tissue, Membrane Tween 20 Ultracentrifugation uranyl acetate Yeast, Dried
2
Cryopreservation of Neural Tissues
Cited 8 times
A fundamental hurdle in freezing biological tissues is the formation of ice crystals that distort or damage structures. Cryoprotectants can damage structures, such as membranes, or distort cells by shrinking them, but cryoprotectants can be avoided if the heat can be removed in one millisecond or less. To date it has only been possible to achieve these rates of cooling within a few micrometers of the surface in a neural tissues (Heuser and Reese, 1981 (link)), limiting investigation almost entirely to disassociated cultures or surfaces of acute brain slices. High pressure freezing (HPF) methods (Moore, 1987 ), which inhibit formation of ice crystals by exertion of large pressures at the moment of freezing, can extend the depth of freezing to 100–300 μm or more and, when optimally implemented, can well preserve structures down to atomic resolutions (Dubochet, 2007 (link)). However, in practice, results may be capricious and current HPF machines limit the size of the tissue to one to three millimeters. Other HPF methods to freeze reliably larger volumes of tissue are under development. The heat transfer rate for HPF is generally lower than that of rapid slam freezing (Heuser and Reese, 1981 (link)), so HPF might not be the best choice for capturing very fast dynamic events. Despite of these challenges, HPF would be the method of choice for preparing nerve tissues for many future EM tomography investigations where the emphasis is focused on macromolecular organization in neurons. Here we present an example of a system for tomography based on HPF and freeze-substitution of dissociated neuronal cultures (Fig. 1).
Chen X., Winters C.A, & Reese T.S. (2008). Life Inside a Thin Section: Tomography. The Journal of Neuroscience, 28(38), 9321-9327.
Publication 2008
Biopharmaceuticals Brain Cardiac Arrest Cells Cryoprotective Agents Electron Microscope Tomography Freeze Substitution Nerve Tissue Neurons Pressure Tissue, Membrane Tissues Tomography
3
Drosophila Brain Tissue Preparation for FIB-SEM
Cited 7 times
Two different methods were used to prepare Drosophila brain tissue imaged by FIB-SEM. For one approach, the head of a 5-day-old adult female CantonS G1xw1118 Drosophila was cut into 200 μm slices with a Leica VT1000 microtome in 2.5% glutaraldehyde and 2.5% paraformaldehyde, in 0.1 M cacodylate at pH 7.3. The vibratome slice was fixed for a total of 10–15 min, transferred to 25% aqueous bovine serum albumin for a few minutes, and then loaded into a 220 μm deep specimen carrier and high-pressure frozen in a Wohlwend HPF Compact 01 High-Pressure Freezing Machine (Wohlwend Gmbh). The brain was then freeze-substituted in a Leica EM AFS2 system in 1% osmium tetroxide, 0.2% uranyl acetate and 5% water in acetone with 1% methanol, for three more days (Takemura et al., 2015 (link)). At the end of freeze-substitution, the temperature was raised to 22°C and tissues was rinsed in pure acetone, then infiltrated, and embedded in Durcupan epoxy resin (Fluka).
Alternatively, whole Drosophila brains were fixed in 2.5% formaldehyde and 2.5% glutaraldehyde in 0.1 M phosphate buffer at pH 7.4 for 2 hr at 22°C. After washing, the tissues were post-fixed in 0.5% osmium tetroxide in ddH2O for 30 min at 4°C. After washing and en bloc staining with 0.5% aqueous uranyl acetate for 30 min, a Progressive Lowering Temperature (PLT) procedure started from 1°C when the tissues were transferred into 10% acetone. The temperature was progressively decreased to −25°C, while the acetone concentration was gradually increased to 97%. The tissue was fixed in 1% osmium tetroxide and 0.2% uranyl acetate in acetone for 32 hr at −25°C. After PLT and low-temperature incubation, the temperature was increased to 22°C, and tissues were rinsed in pure acetone, then infiltrated, and embedded in Poly/Bed 812 (Luft formulation).
Xu C.S., Hayworth K.J., Lu Z., Grob P., Hassan A.M., García-Cerdán J.G., Niyogi K.K., Nogales E., Weinberg R.J, & Hess H.F. (2017). Enhanced FIB-SEM systems for large-volume 3D imaging. eLife, 6, e25916.
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Publication 2017
Acetone Brain Buffers Cacodylate Cold Temperature Drosophila Durcupan Epoxy Resins Focused Ion Beam Scanning Electron Microscopy Formaldehyde Freeze Substitution Freezing Glutaral Head Methanol Microtomy Osmium Tetroxide paraform Phosphates Polybed 812 Pressure Serum Albumin, Bovine Tissues uranyl acetate Woman
4
High-Pressure Freezing for Ultrastructural Analysis
Cited 7 times
High-pressure freezing and freeze substitution was conducted as described previously for morphometric analysis (Weimer, 2006 ) with modifications. In short, about 10 young adult hermaphrodites were loaded into the 100 μm deep well of the freezing chamber (Specimen Carriers Type A (100 μm) and B (0 μm), Bal-Tec AG, Liechtenstein, acquired by Leica Microsystems, Wetzlar, Germany) filled with a mixture of 10% BSA and pelleted OP50 bacteria (ratio: 20:100, v/v) and processed by an EM HPM100 (Leica Microsystems, Wetzlar, Germany) at a freezing speed > 20 000 K/s and a pressure > 2000 bar. High-pressure frozen worm/bacteria pellets were directly transferred in liquid nitrogen to an EM AFS2 freeze substitution system (Leica Microsystems, Wetzlar, Germany). Freeze chambers were opened to allow better penetration during freeze substitution. Samples were incubated in 0.1% tannic acid, 0.5% glutaraldehyde in anhydrous acetone at -90 °C for 96 h, washed 4 times for 1 h with anhydrous acetone at -90 °C and fixed in 2% OsO4 in anhydrous acetone at -90 °C for 28 h, then temperature was ramped for 14 h to -20 °C, incubated 16 h at -20 °C, ramped again to 4 °C in 4 h and immediately washed with anhydrous acetone at 4 °C 4 times at 0.5 h intervals. Then the temperature was gradually raised to 20 °C in 1h and samples were subsequently transferred for embedding in a series of freshly prepared araldite solutions at increasing concentrations (50% araldite in acetone for 3 h at room temperature, 90% araldite in acetone overnight at 4 °C followed by 2 times pure araldite at room temperature). Araldite infiltrated samples were polymerized for 48 h at 60 °C.
Stigloher C., Zhan H., Zhen M., Richmond J, & Bessereau J.L. (2011). The Presynaptic Dense Projection of the Caenorhabiditis elegans Cholinergic Neuromuscular Junction Localizes Synaptic Vesicles at the Active Zone through SYD-2/Liprin and UNC-10/RIM-Dependent Interactions. The Journal of Neuroscience, 31(12), 4388-4396.
Publication 2011
Acetone araldite Bacteria Freeze Substitution Freezing Glutaral Helminths Hermaphroditism Nitrogen Pellets, Drug Pressure Tannins Young Adult
5
Ultrastructural Analysis of Chick Tendons and Mouse Tails
Cited 7 times
Freshly dissected chick metatarsal tendons were cut into 3-mm lengths and frozen to −196°C using an EM PACT high pressure freezer (Leica). Freeze substitution for ultrastructure was performed using an AFS system (Leica), starting at –90°C in 2% wt/vol osmium tetroxide in actone, going through pure acetone at −50°C and ending in several changes of Spurr's resin (Spurr, 1969 (link)). at 20°C. Polymerization in fresh resin was then performed at 60°C for 24 h. Freeze substitution for immunolabeling was performed using an AFS system (Leica) using pure acetone at –90°C, pure ethanol at −50°C in ethanol, and ending in several changes of HM20 Lowicryl resin at −50°C. UV polymerization in fresh resin was then performed at –50°C for 48 h and continued at 20°C for 48 h.
Embryonic mouse tails were fixed in 2% glutaraldehyde in 100 mM phosphate buffer, pH 7.0, for 30 min at RT. The tails were then diced and fixed for 2 h at 4°C in fresh fixative. After washing in 200 mM phosphate buffer they were fixed after in 1% glutaraldehyde and 1% OsO4 in 50 mM phosphate buffer, pH 6.2, for 40 min at 4°C. After a rinse in distilled water they were en bloc stained with 1% aqueous uranyl acetate for 16 h at 4°C, dehydrated and embedded in Spurrs' resin.
Ultra-thin sections for normal transmission electron microscopy were collected on uncoated copper 200 grids, serial sections for 3-D reconstruction on formvar-coated copper 1,000 μm slot grids (stabilized with carbon film) and ultra-thin sections (∼60 nm) for immunolabeling on formvar-coated nickel 400 grids. A postembedding labeling technique was used to detect type I collagen using a rabbit anti–chicken collagen-I antibody (Biodesign International) at a dilution of 1:500 followed by a gold-conjugated goat anti–rabbit antibody (British Biocell International) at a dilution of 1:200. All sections were subsequently stained with uranyl acetate and lead citrate, and examined using either a JEOL 1200EX, Philips EM 400, or Philips BioTwin transmission electron microscope. Images were recorded on 4489 film (Kodak) and scanned using an Imacon Flextight 848 scanner (Precision Camera & Video). Images from EM serial sections were aligned and reconstructed in IMOD for Linux (Kremer et al., 1996 (link)) and visualized using OpenSynu for Linux (Hessler et al., 1992 (link)).
Canty E.G., Lu Y., Meadows R.S., Shaw M.K., Holmes D.F, & Kadler K.E. (2004). Coalignment of plasma membrane channels and protrusions (fibripositors) specifies the parallelism of tendon. The Journal of Cell Biology, 165(4), 553-563.
Publication 2004
Acetone Antibodies, Anti-Idiotypic Buffers Carbon Chickens Citrates Collagen Type I Copper Embryo Ethanol Fixatives Formvar Freeze Substitution Freezing Glutaral Goat Gold IMod Metatarsal Bones Mice, House Microtomy Nickel Osmium Tetroxide Phosphates Polymerization Pressure Rabbits Reconstructive Surgical Procedures Resins, Plant spurr resin Tail Technique, Dilution Tendons Transmission Electron Microscopy uranyl acetate

Most recents protocols related to «Freeze Substitution»

1
High-pressure Freezing TEM Sample Preparation of Lacrimia sp.
The sample preparation of Lacrimia sp. YPF1808 by the high-pressure freezing technique followed the protocol for TEM sample preparation. After freeze-substitution, the samples were subsequently stained with 1% thiocarbohydrazide in 100% acetone for 1.5 h, 2% OsO4 in 100% acetone for 2 h at room temperature, and 1% uranyl acetate in 100% acetone overnight at 4°C. After every staining step, the samples were washed 3 times with 100% acetone for 15 min. Samples were then infiltrated with 25%, 50%, or 75% acetone-resin mixture for 2 h at each step, and finally infiltrated in 100% Hard Resin Plus 812 (EMS) overnight and polymerized at 62°C for 48 h. Resin-embedded blocks were trimmed and imaged using an Apreo SEM equipped with a VolumeScope (Thermo Fisher Scientific, Germany). Serial images were acquired at 3.5 keV, 50 pA, 40 Pa with a resolution of 6 nm, 100-nm slice thickness, and dwell time per pixel of 4 μs. Image data were processed in Microscopy Image Browser v2.702 (65 (link)) and Amira v2020.2. The resin-embedded blocks were also collected in the form of 1-μm-thick sections on a silicon wafer and analyzed by SEM-EDX (Magellan 400L system, as described above).
Based on volumetric data, we calculated the percentage of increase in cell density based on measured volumes of crystals compared to the theoretical crystal-free cells of the same volume and reported average theoretical density of 1.07 g·cm−3 (37 ).
Pilátová J., Tashyreva D., Týč J., Vancová M., Bokhari S.N., Skoupý R., Klementová M., Küpper H., Mojzeš P, & Lukeš J. (2023). Massive Accumulation of Strontium and Barium in Diplonemid Protists. mBio, 14(1), e03279-22.
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Publication 2023
Acetone Cells Freeze Substitution Freezing Microscopy Pressure Resins, Plant Silicon thiocarbohydrazide uranyl acetate
2
Transmission Electron Microscopy Sample Preparation
The protocol for the basic sample preparation of all kinds of electron microscopy approaches listed here is described in detail elsewhere (61 (link)). We used it with minor modifications, as stated below. Cell pellets were transferred to specimen carriers and immediately frozen in the presence of 20% (wt/vol) bovine serum albumin solution using a high-pressure freezer (Leica EM ICE, Leica Microsystems, Austria). Freeze substitution was performed in the presence of 2% osmium tetroxide diluted in 100% acetone at −90°C. After 96 h, specimens were warmed to −20°C at a step of 5°C/h. After another 24 h, the temperature was increased to 3°C (3°C/h). At room temperature, samples were washed in acetone and infiltrated with 25%, 50%, and 75% acetone/resin mixture for 1 h at each step. Finally, samples were infiltrated in 100% resin and polymerized at 60°C for 48 h. Semithin (250 nm) and ultrathin (70 nm) sections were cut using a diamond knife, placed on copper grids, and stained with uranyl acetate and lead citrate. TEM micrographs were taken with a Mega View III camera (SIS) using a JEOL 1010 TEM operating at an accelerating voltage of 80 kV.
For TEM-EDX, 10 μL of pelleted L. lanifica cells was spread over a holey carbon-coated copper grid, washed twice with 10 μL of distilled water in order to reduce the sea salts from the culture medium, and allowed to dry by evaporation at ambient temperature. Semithin sections of resin-infiltrated blocks of N. karyoxenos were prepared as stated above. For the identification of the crystalline phase, sections were studied by TEM on an FEI Tecnai 20 system (LaB6, 120 kV) equipped with an Olympus SIS charge-coupled-device camera Veleta (2,048 by 2,048 pixels) and an EDAX windowless EDX detector Apollo XLTW for elemental analysis. The diffraction data were collected by means of 3D electron diffraction (ED) (62 (link)). The data processing was carried out using PETS software (63 (link)). Structure solution and refinement were performed in the computing system Jana2006 (64 (link)).
Pilátová J., Tashyreva D., Týč J., Vancová M., Bokhari S.N., Skoupý R., Klementová M., Küpper H., Mojzeš P, & Lukeš J. (2023). Massive Accumulation of Strontium and Barium in Diplonemid Protists. mBio, 14(1), e03279-22.
Full text: Click here
Publication 2023
Acetone Carbon Cells Citrates Copper Culture Media Diamond Electron Microscopy Electrons Freeze Substitution Freezing L Cells Medical Devices Osmium Tetroxide Pellets, Drug Pets Pressure Resins, Plant Salts Serum Albumin, Bovine uranyl acetate
3
Immunogold Labeling and Electron Microscopy
High-pressure freezing (HPF), freeze substitution, ultra-thin sectioning, and imaging were performed as previously described (64 (link)). Immunogold labeling for EM was performed with anti-GFP at optimized concentrations, then with goat anti-rabbit 10 nm gold-coupled secondary antibodies at 1:40 dilution. TEM examination was performed with a Hitachi H-7650 transmission electron microscope with a charge-coupled device (CCD) camera operating at 80 kV (Hitachi High-Technologies). ET, 3D reconstruction, and modeling were performed as described previously (63 (link), 64 (link)).
Li B., Niu F., Zeng Y., Tse M.K., Deng C., Hong L., Gao S., Lo S.W., Cao W., Huang S., Dagdas Y, & Jiang L. (2023). Ufmylation reconciles salt stress-induced unfolded protein responses via ER-phagy in Arabidopsis. Proceedings of the National Academy of Sciences of the United States of America, 120(5), e2208351120.
Publication 2023
Anti-Antibodies Freeze Substitution Goat Gold Medical Devices Pressure Rabbits Reconstructive Surgical Procedures Technique, Dilution Transmission Electron Microscopy
4
Freeze Substitution Protocol for Muscle Samples
The samples were transferred to an EM AFS2 freeze substitution system (Leica Microsystems GmbH) and incubated using the following FS protocol either with ethanol absolute or acetone as solving agents: For 48 h, the muscle filets were incubated in ethanol absolute (Th. Geyer GmbH & Co. KG, Renningen, Germany) or anhydrous acetone (AppliChem GmbH, Darmstadt, Germany) at −90 °C. To efficiently remove residual water, the solution was refreshed after 24 h. After 48 h, the temperature was gradually raised to −60 °C within 15 h. At −60 °C, the samples were incubated with 2% PFA (AppliChem GmbH) in ethanol absolute [21 ] or anhydrous acetone, respectively, for 8 h. Within 18 h, the temperature was gradually raised to −30 °C, at which the samples were repeatedly washed with 2% PFA in either ethanol or acetone. After 17 h at −30 °C, the temperature was increased to 4 °C within 6 h. At 4 °C, the samples were washed with ethanol absolute/anhydrous acetone four times within 4 h, followed by a rehydration described as follows: In total, eight washings steps of 10 min each were performed with ethanol absolute/acetone in phosphate buffered saline (PBS) including 95%, 90%, 80%, 70%, 50%, 30% ethanol absolute/anhydrous acetone in PBS, and two rinses in 100% PBS.
Mrestani A., Lichter K., Sirén A.L., Heckmann M., Paul M.M, & Pauli M. (2023). Single-Molecule Localization Microscopy of Presynaptic Active Zones in Drosophila melanogaster after Rapid Cryofixation. International Journal of Molecular Sciences, 24(3), 2128.
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Publication 2023
Absolute Alcohol Acetone Ethanol Freeze Substitution Muscle Tissue Phosphates Rehydration Saline Solution
5
Ultrastructural Analysis of Extracellular Space
Mice were injected with 10% chloral hydrate and transcardially perfused with 0.1 M sodium cacodylate buffer followed by ice-cold 2% PFA and post-fixed with 0.5% PFA at 4 °C. Tissue was sectioned on a vibratome, and freeze substitution and low temperature embedding of the specimens was performed as described previously83 (link),84 (link),85 (link). Slices were cryoprotected by immersion in increasing concentrations of glycerol (from 10% to 30% in PBS) (v/v). Sections were plunged rapidly into liquid propane cooled by liquid nitrogen (−190 °C) in a Universal Cryofixation System KF80 (Reichert-Jung, Vienna, Austria). The samples were immersed in 1.5% uranyl acetate dissolved in anhydrous methanol (−90 °C, 24 h) in a cryosubstitution AFS unit (Leica, Vienna, Austria). The temperature was raised from −90 °C to −45 °C in steps of 4 °C/h. After washing with anhydrous methanol, the samples were infiltrated with Lowicryl HM20 resin (Electron Microscopy Sciences, Fort Washington, PA) at −45 °C. Polymerization with ultraviolet light (360 nm) was performed for 48 h at −45 °C, followed by 24 h at 0 °C. Ultrathin sections (80 nm) were cut with a diamond knife on a Leica UC7 ultramicrotome and mounted on 300 mesh copper grids using a Coat-Quick adhesive pen (Electron Microscopy Sciences). Images (n=10/animal) were taken using a Hitachi 7700 electron microscope (Hitachi High-Technologies Corporation America, Inc.) equipped with a XR81-B-M1-BT-FX, 8 Megapixel digital camera (Advanced Microscopy Techniques, Woburn, MA). Images were then imported into Adobe Photoshop (Adobe, 2022) and the extracellular space was manually scored using a computer tablet. Scoring was done by two independent investigators blinded to experimental conditions. Images were then imported into ImageJ (v1.53f51)86 (link) and the percentage of marked area/total area was calculated.
Cathomas F., Lin H.Y., Chan K.L., Li L., Durand-de Cuttoli R., Parise L.F., Aubry A.V., Muhareb S., Desland F., Shimo Y., Ramakrishnan A., Estill M., Ferrer-Pérez C., Parise E.M., Wang J., Sowa A., Janssen W.G., Costi S., Rahman A., Fernandez N., Swirski F.K., Nestler E.J., Shen L., Merad M., Murrough J.W, & Russo S.J. (2023). Peripheral immune-derived matrix metalloproteinase promotes stress susceptibility. Research Square.
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Publication Preprint 2023
Animals Buffers Cacodylate Cold Temperature Copper Cryopreservation Diamond Electron Microscopy Extracellular Space Fingers Freeze Substitution Glycerin Hydrate, Chloral Immersion Methanol Mice, House Microscopy Nitrogen Polymerization Propane Resins, Plant Sodium Tablet Tissues Ultramicrotomy Ultraviolet Rays uranyl acetate

Top products related to «Freeze Substitution»

1
Em afs2 by Leica
Sourced in Germany
The EM AFS2 is a high-performance automated freeze substitution system designed for the preparation of biological samples for electron microscopy. It provides precise control of temperature, time, and reagent delivery to ensure consistent and reproducible sample preparation.
2
Em pact2 by Leica
Sourced in Germany, United Kingdom
The Leica EM PACT2 is a high-performance freeze-substitution and cryofixation system designed for advanced sample preparation in electron microscopy. It is a versatile instrument that can be used for a wide range of cryogenic applications.
3
Hpm100 by Leica
Sourced in Germany, United States
The HPM100 is a high-performance microscope platform designed for laboratory use. It provides a stable and precise optical system to support various imaging and analysis applications.
4
Em hpm100 by Leica
Sourced in Germany, Austria
The EM HPM100 is a high-pressure freezing machine manufactured by Leica. It is designed to rapidly freeze biological samples under controlled high-pressure conditions.
5
Em afs by Leica
Sourced in Germany, United Kingdom
The EM AFS is a high-performance automated freeze-substitution system developed by Leica for the preparation of biological samples for electron microscopy. It provides a controlled, reproducible, and efficient method for dehydrating and embedding samples while preserving their ultrastructural details.
6
Lowicryl hm20 resin by Polysciences
Sourced in United States, Germany
Lowicryl HM20 resin is a low-viscosity, methacrylate-based embedding medium designed for use in electron microscopy sample preparation. It is formulated to provide good preservation of ultrastructural details and is compatible with various types of specimen staining and labeling techniques.
7
Lowicryl hm20 resin by Electron Microscopy Sciences
Sourced in United States
Lowicryl HM20 is a water-miscible acrylic resin designed for low-temperature embedding of biological samples for electron microscopy. It is compatible with cryogenic sectioning and can be used for immunolabeling procedures.

More about "Freeze Substitution"

Freeze substitution is a powerful technique used in electron microscopy to prepare biological samples for high-resolution analysis.
This cryopreservation method involves rapidly freezing the sample and then gradually replacing the frozen water content with a solvent like acetone or methanol at low temperatures.
This process preserves the delicate ultrastructure of cellular organelles, membranes, and macromolecular complexes, minimizing the artifacts that can occur with traditional chemical fixation.
Freeze substitution is an invaluable tool for researchers investigating the nanoscale structure and function of biological systems.
It allows for the study of fine details that would otherwise be obscured, making it particularly useful for analyzing the intricate workings of cells and their subcellular components.
Optimizing freeze substitution protocols can be a complex process, but the PubCompare.ai platform can help streamline your workflow.
By providing access to relevant literature, preprints, and patents, as well as intelligent comparisons to identify the best methods and prodcuts, PubCompare.ai empowers researchers to make data-driven decisions and achieve reproducible results.
To further enhance your freeze substitution techniques, consider exploring complementary technologies like EM AFS2, EM PACT2, HPM100, EM HPM100, EM AFS, and Lowicryl HM20 resin.
These advanced tools and materials can help you push the boundaries of your research and uncover new insights into the intricate workings of biological systems at the nanoscale level.
OtherTerms: Cryopreservation, Electron Microscopy, Biological Samples, Ultrastructure, Cellular Organelles, Membranes, Macromolecular Complexes, Nanoscale, EM AFS2, EM PACT2, HPM100, EM HPM100, EM AFS, Lowicryl HM20 resin, PubCompare.ai, Data-driven Decision Making, Reproducible Results