Lumbar spinal cord (L1-L3 spinal segments) samples were carefully dissected with the corresponding vertebrae and fixed in 10% neutral buffered formalin (Sigma-Aldrich, HT501128) for 48 h. Tissues were then transferred to decalcifying reagent Immunocal® (StatLab, 1414-32; StatLab Medical Products) for 96 h. Dehydration and paraffin infiltration were performed on a Tissue-Tek VIP 6 Vacuum Infiltration Processor (Sakura Finetek Europe). Following paraffin embedding, 5 μm-thick sections of spinal cord samples were cut and stained with Iba1 antibody, as described for brain sections.
Vip 6 vacuum infiltration processor
Manufactured by Sakura Finetek
The VIP 6 Vacuum Infiltration Processor is a laboratory equipment designed for the controlled infiltration of specimens under vacuum conditions. It facilitates the gradual displacement of air from the specimen's internal structures, enabling the penetration of infiltration media. The device operates using a programmable vacuum system to achieve the desired infiltration process.
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Lab products found in correlation
2 protocols using vip 6 vacuum infiltration processor
1
Immunohistochemical Analysis of Lumbar Spinal Cord
2
Spinal Cord and Sciatic Nerve Tissue Preparation
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Animals were euthanized with an overdose of isoflurane. L1-L3 spinal segments (with the corresponding vertebrae) of lumbar spinal cord were fixed in 10% neutral buffered formalin solution (NBF, Sigma-Aldrich, HT501128) for 48 hours at RT. Sciatic nerve samples were fixed in NBF for 24 hours at RT. Additional de-calcification in Immunocal reagent (StatLab Medical Products, 1414-32) was performed for 72 hours at RT for spinal cord samples. Dehydration and paraffin infiltration were performed on a Tissue-Tek VIP 6 Vacuum Infiltration Processor (Sakura Finetek Europe, Alphen aan den Rijn, the Netherlands). Before paraffin embedding, sciatic nerves were cut in three parts and embedded separately: R1, the branching point into the tibialis muscle, R2/R3, the crush site and R4, the proximal part. Following paraffin embedding 5 μm-thick sections of tissues were mounted on Superfrost Plus Adhesion Microscope Slides (Thermo Fisher Scientific, J1800AMNT). After 48-72 hours at 37°C slides were stored at RT.
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