The following chemicals and drugs were used: propofol-Lipuro (B. Braun Melsungen AG, Melsungen, Germany), diethyl ether for anesthesia (Kuzbassorghim, Kemerovo, Russia), Tween 80 (Merck, Darmstadt, Germany), 2,3,5-triphenyl tetrazolium chloride (TTC) (Sigma, St. Louis, MO, USA), thiopental sodium (Syntez, Kurgan, Russia), 10% neutral formalin, embedding medium Histomix, Vitrogel, Masson Trichrome Stain Kit, picrofuchsin solution for Van Gieson staining (BioVitrum, St. Petersburg, Russia), ethanol (Konstanta-Farm M, Moscow, Russia), and o-xylene (EKOS-1, Moscow, Russia).
Vitrogel
Manufactured by BioVitrum
Sourced in Sao Tome and Principe
Vitrogel is a laboratory equipment designed for cell culture applications. It provides a specialized gel-based substrate for the growth and maintenance of cells in vitro. The core function of Vitrogel is to create a three-dimensional, biomimetic environment that supports the natural behavior and development of cultured cells.
Automatically generated - may contain errors
Lab products found in correlation
Histomix, by BioVitrum (2 mentions)
Tween 80, by Merck Group (1 mentions)
2 3 5 triphenyltetrazolium chloride ttc, by Merck Group (1 mentions)
Propofol lipuro, by B. Braun (1 mentions)
Axioimager a1 light microscope, by Zeiss (1 mentions)
Axiocam mrc5 camera, by Zeiss (1 mentions)
Histostar workstation, by Thermo Fisher Scientific (1 mentions)
Perfection v600 scanner, by Epson (1 mentions)
Microm stp 120, by Thermo Fisher Scientific (1 mentions)
Finesse me microtome, by Thermo Fisher Scientific (1 mentions)
4 protocols using vitrogel
1
Histological Analysis of Tissue Injury
2
Histological Analysis of Teratoma Tissue
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Excised teratomas were washed in PBS and fixed for 24 h at RT in two types of fixators: Bouin’s fluid and 10% formalin. After performing Bouin’s fixation, the teratomas were washed in several changes of 70% alcohol until the solution was lightened. After the formalin fixation, teratomas were washed in running water for 10 min and transferred to 70% alcohol. Specimens were dehydrated in an ethanol series and isobutanol:paraffin series and then embedded in paraffin (McCormick Scientific, now Leica Biosystems, Nussloch, Germany). For each teratoma, blocks of approximately 5 × 5 mm were used for analysis. Paraffin sections were washed in xylene, rehydrated using an ethanol series (100–70%), and then washed with water. Next, sections were incubated in hematoxylin for 6 min, washed with water, and incubated with eosin for 3 min. After washing and dehydration, sections were mounted in Vitrogel (BioVitrum, St-Petersburg, Russia) for further analysis.
3
Microscopic Analysis of Phloem and Xylem
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Three samples of phloem and xylem tissues from each tree were analyzed. Sample preparation for microscopy was conducted as described previously [26 (link)]. The samples were fixed in glutaraldehyde, dehydrated in a series of alcohols of rising concentrations, and embedded in an Araldite-Embed-812 mixture following the published technique [42 (link)]. Cross sections 2 µm thick were cut with an Ultrotome IV (LKB, Bromma, Sweden) and stained with a 1% aqueous solution of safranin. Permanent slides were made using synthetic mounting medium Vitrogel (BioVitrum, St. Petersburg, Russia). Microscopic analysis was carried out under an AxioImager A1 light microscope (Carl Zeiss, Jena, Germany) equipped with an ADF PRO03 camera. Images were processed with ADF Image Capture software (ADF Optics Ningbo, China). Anatomical measurements were made following the available guidelines using panoramic cross sections with an area of 7–10 mm2 [43 ,44 (link),45 (link)].
4
Assessing Lung Pathology in Mice
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Lungs from each mouse were fixed in 10% neutral buffered formalin (Soluformtm, JLS-Chemical, Russia) at +4°C, dehydrated in isoprepe (BioVitrum, Russia) using Microm STP 120 (Thermo Scientific, USA), and embedded in HISTOMIX (BioVitrum, Russia) using HistoStar workstation (Thermo Scientific, USA). 5-μm thick sections (10 per mouse) were cut using a Finesse ME+ microtome (Thermo Scientific, USA), stained with haematoxylin and eosin and mounted in Vitrogel (all BioVitrum, Russia). Pictures were obtained either using an Epson Perfection V600 scanner (9600 dpi) or using an Imager. Z1 microscope with an AxioCam MRc 5 camera (all ZEISS, Germany) at 20х and 63х magnification. Pathological changes were assessed using three parameters: (1) the acute lung injury (ALI) score, showing mostly the immunopathology in the lung parenchyma (from 0 to 1); (2) the peribronchiolar infiltration score, estimating the degree of inflammation of the tissue around the bronchioles (from 0 to 5); and (3) the perivascular infiltration score, estimating the degree of inflammation of the tissue around vessels (from 0 to 5). The exact parameters of tissue scoring systems are provided in the Supplementary Materials (p. 9).
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