Pp3010t cryo sem preparation system

Manufactured by Quorum Technologies

Sourced in United Kingdom

The PP3010T Cryo-SEM Preparation System is a laboratory instrument designed for the preparation of samples for cryo-scanning electron microscopy (Cryo-SEM) analysis. The core function of the PP3010T is to prepare samples by rapidly freezing them and transferring them to the SEM chamber while maintaining their cryogenic state.

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Lab products found in correlation

Jsm 7100f sem, by JEOL (3 mentions) Dm2500 microscope, by Leica (3 mentions) Jsm 7100f, by JEOL (2 mentions) Nis elements software, by Nikon (1 mentions) A1r confocal microscope, by Nikon (1 mentions) Css450, by Linkam (1 mentions) Agar sputter coater b7340, by Agar Scientific (1 mentions) Su8010 sem, by Hitachi (1 mentions) Mc170 hd color camera, by Leica (1 mentions)

Close spelling variants of this product

Quorum PP3010T Cryo-FIB/SEM Preparation System PP3010T

8 protocols using pp3010t cryo sem preparation system

Cryo-SEM Imaging and Structural Analysis

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A small amount of sample (1 – 2 g) was placed into a slot on a stub with rivets and cryo-vitrified with liquid nitrogen rapidly to prevent the formation of large ice crystals. After being transferred onto a cryo-stage (at −160 °C) of the PP3010T Cryo-SEM Preparation System (Quorum Technologies, UK), samples were fractured and subsequently the frozen water was sublimated at −90 °C under controlled vacuum conditions. After 15 min, samples were sputtered with platinum using argon gas to prevent charging during electron beam targeting before being transferred on to the JSM 7100F SEM stage (JEOL ltd., Japan) at −160 °C and scanned at a magnification of 1 × 10⁴ (Patel et al., 2015 (link)).
All cryo-SEM images are segmented using Otsu thresholding by the open-source ImageJ software (National Institutes of Health, USA) in order to extract the network information. Removal of outliers was performed to remove noise and unwanted small objects from the structure. We segmented images using watershed by pixel flooding for pore partitioning and labelled those segmented meshes and watershed ridges for fibril diameter and mesh size calculation. Three images were used for each sample and ten different segments on each image were randomly measured.

Song Q., Wu L., Li S., Zhao G., Cheng Y, & Zhou Y. (2022). Aggregation of konjac glucomannan by ethanol under low-alkali treatment. Food Chemistry: X, 15, 100407.

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Microstructure Analysis of Materials

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The microstructure was studied using Leica DM2500 microscope (Leica Microsystems, Belgium) under normal and polarized light. For confocal microscopy, samples were imaged using a Nikon A1R confocal microscope (Nikon Instruments Inc., USA). Excitation was performed by means of a 488 nm Ar laser and fluorescence was detected through a 525/50 bandpass filter. Images were acquired and processed with Nikon NIS Elements software. For cryo-SEM, samples were placed in the slots of a stub, plunge-frozen in liquid nitrogen, and transferred into the cryo-preparation chamber (PP3010T Cryo-SEM Preparation System, Quorum Technologies, UK) where it was freeze-fractured and subsequently sputter-coated with Pt and examined in JEOL JSM 7100F SEM (JEOL Ltd., Tokyo, Japan). For water containing samples, sublimation step was included to get rid of water.

Patel A.R, & Dewettinck K. (2015). Comparative evaluation of structured oil systems: Shellac oleogel, HPMC oleogel, and HIPE gel. European Journal of Lipid Science and Technology, 117(11), 1772-1781.

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Microstructure Analysis of Emulsions

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Optical and cryo-scanning electron microscopy (cryo-SEM) was utilized to study the microstructure of the samples. Optical microscopy was done on Leica DM2500 microscope (Leica Micro-systems, Belgium). For cryo-SEM, samples of the emulsions were placed in the slots of a stub, plunge-frozen in slush nitrogen and transferred into the cryo-preparation chamber (PP3010T cryo-SEM Preparation System, Quorum Technologies, UK) where they were freeze-fractured, sublimated for 20 min and subsequently sputter-coated with Pt and examined by a JEOL JSM 7100F SEM (JEOL Ltd, Tokyo, Japan).

Gao Z., Daneva A., Salanenka Y., Van Durme M., Huysmans M., Lin Z., De Winter F., Vanneste S., Karimi M., Van De Velde J., Vandepoele K., Van de Walle D., Dewettinck K., Lambrecht B.N, & Nowack M.K. (2018). KIRA1 and ORESARA1 terminate flower receptivity by promoting cell death in the stigma of Arabidopsis. Nature plants, 4(6), 365-375.

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3D Morphology of Fat Crystal Flocs

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Cryo-scanning electron microscopy (cryo-SEM) was applied to unravel the 3D morphology of the fat crystal flocs. The samples were crystallized following the protocols described above. Crystallization toward 20 °C or 25 °C was performed on the PE120 Peltier system of the PLM, without cover glass. Crystallization toward 0 °C was performed in a shear cell CSS450 equipped with liquid nitrogen cooling system (Linkam, Redhill, UK). After one hour of isothermal time, the sample (about the volume of a droplet) was transferred to a pre-tempered aluminum cryo-SEM stub covered with carbon tape. The sample was de-oiled by dripping 1 mL of isobutanol on top and letting it evaporate at the crystallization temperature. This was repeated four times. Lastly, 1 mL of acetone was dripped on top and also allowed to evaporate. After vitrification by means of a nitrogen slush, the sample was transferred into the cryo-preparation chamber (PP3010T Cryo-SEM Preparation System, Quorum Technologies, Lewes, UK) under vacuum and conditioned at −140 °C. All samples were sublimated for 45 min at −70 °C, sputter-coated with platinum for 90 s and visualized with a cryo-SEM JEOL JSM 7100F (JEOL Ltd., Tokyo, Japan). The SEM stage had a temperature of −140 °C, and the electron beam had an accelerated voltage of 3 keV.

De Witte F., Penagos I.A., Van de Walle D., Skirtach A.G., Dewettinck K, & Van Bockstaele F. (2024). From Nucleation to Fat Crystal Network: Effects of Stearic–Palmitic Sucrose Ester on Static Crystallization of Palm Oil. Foods, 13(9), 1372.

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Visualizing Native and Processed Wheat Bran

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Native, pre-digested and fermented wheat bran samples were visualized using cryo-SEM and desktop SEM. For the purpose of SEM microscopy, the bran samples were chemically dried with hexamethyldisilazane (HMDS) as described by Araujo et al. (2003) (link). After complete evaporation of the HMDS, samples were mounted on an aluminum pin (diameter: 12 mm) using double sided carbon tape and subsequently gold sputtered for 45 s at 30 mA (Agar Sputter Coater B7340, Agar Scientific, UK). Images were collected using a Phenom Pro X SEM microscope (Phenom-World B.V., the Netherlands) with a beam intensity of 10 keV.
As an alternative to SEM microscopy, samples were also visualized via cryo-SEM using a Jeol JSM 7100F scanning electron microscope (JEOL Ltd, Tokyo, Japan). A small amount of wheat bran was placed on a sticky carbon surface mounted on an aluminium stub, vitrified in a nitrogen slush and transferred under vacuum conditions into the cryo-preparation chamber (PP3010T Cryo-SEM Preparation System; Quorum Technologies, Lewes, UK) conditioned at −140 °C. Subsequently, the sample was sublimated for 20 min at −70 °C to remove frost artefacts, sputter-coated with platinum using argon gas, transferred to the SEM stage at −140 °C and electron beam targeted at 3 keV.

De Paepe K., Verspreet J., Rezaei M.N., Hidalgo Martinez S., Meysman F., Van de Walle D., Dewettinck K., Raes J., Courtin C, & Van de Wiele T. (2019). Isolation of wheat bran-colonizing and metabolizing species from the human fecal microbiota. PeerJ, 7, e6293.

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Cryo-SEM Imaging of Immobilized Gels

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SU8010 SEM (Hitachi Co., Ltd., Tokyo, Japan) was applied to obtain cryo-SEM images. Various gels were immobilized in a copper holder and then frozen and subjected to liquid nitrogen slush, which was quickly delivered to a cryo-preparation chamber (PP3010T cryo-SEM preparation system, Quorum Technologies, Hertfordshire, UK) under vacuum. Various samples were further subjected to freeze-fracture, sublimation (−60 °C, 40 min), and Pt spraying (10 mV, 60 s). Finally, cryo-SEM images were captured at a 10.0 kV accelerating voltage.

Yan J.N., Nie B., Zhang Z.J., Gao L.Y., Lai B., Wang C., Zhang L.C, & Wu H.T. (2023). Monovalent Salt and pH-Stimulated Gelation of Scallop (Patinopecten yessoensis) Male Gonad Hydrolysates/κ-Carrageenan. Foods, 12(19), 3598.

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Wax Crystal Microstructure Visualization

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The microstructure of wax crystals was observed under normal and polarized light (PLM) using a Leica DM2500 microscope (Wetzlar, Germany) equipped with a Leica MC170 HD color camera. For cryo-scanning electron microscopy (cryo-SEM), deoiling of organogel samples was carried out using butanol to remove the surface liquid oil in order to visualize the wax crystals. Precisely, the deoiling was carried out in two different ways: (1) A known quantity of organogel was weighed in a glass vial followed by addition of butanol (at a gel:butanol ratio of 1:50 w/w approximately), which led to the collapse of the gel structure and resultant sedimentation of the crystalline fraction. After overnight storage, the supernatant liquid was decanted to collect the sediment, which was then placed on the sample holder. (2) The organogel sample was directly placed on a specialized stub (sample holder with grooves) followed by a dropwise addition of butanol to remove the liquid oil. The stub was left for overnight drying in inverted position to drain out all the solvent. The stub was then plunge-frozen in liquid nitrogen and transferred into the cryo-preparation chamber (PP3010T Cryo-SEM Preparation System, Quorum Technologies, UK), where it was freeze-fractured and subsequently sputter-coated with Pt and examined in a JEOL JSM 7100F SEM (JEOL Ltd., Tokyo, Japan).

Patel A.R., Babaahmadi M., Lesaffer A, & Dewettinck K. (2015). Rheological profiling of organogels prepared at critical gelling concentrations of natural waxes in a triacylglycerol solvent. Journal of agricultural and food chemistry, 63(19).

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SEM Visualization of Wheat Bran Samples

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As an alternative to SEM microscopy, samples were also visualised using a Jeol JSM 7100F scanning electron microscope (JEOL Ltd, Tokyo, Japan). A small amount of wheat bran was placed on a sticky carbon surface mounted on an aluminium stub, vitrified in a nitrogen slush and transferred under vacuum conditions into the cryopreparation chamber (PP3010T Cryo-SEM Preparation System, Quorum Technologies, UK) conditioned at -140°C. Subsequently, the sample was sublimated for 20 min at -70°C to remove frost artefacts, sputter-coated with platinum using argon gas, transferred to the SEM stage at -140°C and electron beam targeted at 3 keV.

De Paepe K ., Verspreet J ., Rezaei MN ., Martinez SH ., Meysman F ., Van de Walle D ., Dewettinck K ., Courtin CM , & Van de Wiele T . (2019). Modification of wheat bran particle size and tissue composition affects colonisation and metabolism by human faecal microbiota. Food & function, 10(1).

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