The obtained material was fixed in neutral 10% formalin solution. The bone tissue was decalcified in an acid-free solution. Standard histologic diagnosis was performed using the Excelsior ES apparatus (Thermo Scientific, USA). After the diagnosis, paraffin blocks were made using the HistoStar embedding workstation (Thermo Scientific). 4–6 μm thick sections were obtained using the Microm HM 325 microtome (Thermo Scientific), stained with hematoxylin and eosin using the Gemini AS staining station (Thermo Scientific). For morphometric processing and creation of a video archive of the obtained material, the Leica 2500 microscope (Leica Biosystems, Germany) was used, field lens ×4, ×10, ×20, ×40, ×100, eyepiece lens ×10.
Microm hm 325 microtome
Manufactured by Thermo Fisher Scientific
The Microm HM 325 is a manual rotary microtome designed for sectioning a wide range of samples for microscopic analysis. It features a sturdy construction, a precise specimen feed mechanism, and a range of specimen orientations to accommodate various specimen types.
Automatically generated - may contain errors
Lab products found in correlation
Excelsior es apparatus, by Thermo Fisher Scientific (2 mentions)
Gemini as staining station, by Thermo Fisher Scientific (2 mentions)
Histostar embedding workstation, by Thermo Fisher Scientific (1 mentions)
Cm1850 cryostat, by Leica (1 mentions)
Histostar, by Thermo Fisher Scientific (1 mentions)
Gemini as, by Thermo Fisher Scientific (1 mentions)
4 protocols using microm hm 325 microtome
1
Histological Preparation and Analysis of Bone Tissue
2
Tissue Preparation for Histological Analysis
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For fresh frozen tissues, 3mm sections of the liver large lobe were embedded in the OCT compound (TissueTek) following standard protocols, and 18m-thick sections were cut using a Leica CM1850 cryostat and stored at -80 °C.
For paraffin-embedded tissues, 3mm sections of the liver large lobe were fixed in 4% neutralbuffered formalin (VWR Chemicals) at room temperature (RT) overnight, and stored in 70% ethanol at 4°C. Fixed tissues were processed using standard protocols and embedded in paraffin wax. Three-µm-thick sections were cut using a Thermo Scientific Microm HM325 microtome, dried at 37 °C overnight and stored at 4 °C.
For paraffin-embedded tissues, 3mm sections of the liver large lobe were fixed in 4% neutralbuffered formalin (VWR Chemicals) at room temperature (RT) overnight, and stored in 70% ethanol at 4°C. Fixed tissues were processed using standard protocols and embedded in paraffin wax. Three-µm-thick sections were cut using a Thermo Scientific Microm HM325 microtome, dried at 37 °C overnight and stored at 4 °C.
3
Histological Evaluation of Intestinal Tissue
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At the end of the experiment, regions of interest of the intestinal wall studied with OCTA were excised and taken for blinded histological evaluation. Tissues were fixed in 10% formalin, sent to standard histological wiring, and embedded in paraffin blocks (HistoStar, Thermo Scientific, Waltham, MA, USA). Serial histological sections 4–6 microns thick (Microm HM 325 microtome, Thermo Scientific) were stained with hematoxylin and eosin (Gemini AS, Thermo Scientific). The presence of hemorrhages and mechanical ruptures in the layers of the intestinal wall which could be the result of using a vacuum stabilizer were assessed by an independent histopathologist.
4
Histological Examination of Bone Regeneration
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Sampling for histological examination was carried out by carefully separating muscles from bones followed by isolating bone segments 1–1.5 cm long including the bone regeneration areas. This bone material was fixated in 10% buffered formalin solution for three days and then decalcified in Trilon-V solution (Khimreaktiv, Russia). The standard histological processing was performed using an Excelsior ES apparatus (Thermo Fisher Scientific, USA). After that, paraffin blocks were prepared using a HistoStar filling station (Thermo Fisher Scientific). Sections 4–6 μm thick were obtained with the help of a Microm HM 325 microtome (Thermo Fisher Scientific) and stained with hematoxylin and eosin using a Gemini AS staining station (Thermo Fisher Scientific). Microscopic examination was carried out with a Leica DM2500 light-optical microscope (Leica Microsystems) under ×100 or ×200 magnifications.
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