Tumor tissue analogs were collected and fixed in 10% formalin for 2 h, washed with PBS and dehydrated twice with 50 and 70% ethanol for 15 min each. The dehydrated TTA were transferred to the cryomold (Thermo Fisher Scientific Inc., Waltham, MA) and embedded in HistoGel™ (Thermo Scientific™, Kalamazoo, MI) liquefied by heating at 60° ± 5°C. The TTA containing Cryomolds® were subsequently solidified ice for 10 min. The HistoGel blocks with TTA were wrapped within a piece of Bio-Wrap™ (Leica Biosystems, Buffalo Grove, IL) and placed into tissue biopsy cassette (Thermo Fisher Scientific) for processing. Tissue biopsy cassettes containing HistoGel blocks were dehydrated by grades of alcohol and then cleaned with xylene and impregnated with paraffin. Processed HistoGel blocks were then removed from the Bio-Wrap and placed in wax (Paraplast X-TRA®, Sigma-Aldrich) to prepare paraffin blocks. Sections (4–5 μm) were cut with a microtome (Leica Biosystems).
Bio wrap
Manufactured by Leica
Sourced in United States
Bio-Wrap is a laboratory equipment product designed for wrapping and protecting sensitive samples and materials. It serves as a protective barrier to ensure the integrity and safety of the enclosed contents.
Automatically generated - may contain errors
2 protocols using bio wrap
1
Formalin-Fixed Tissue Analog Embedding
2
Paraffin Embedding of 3D Spheroids
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The MTS were sectioned using a microtome (LEICA, RM2125RTF). Firstly, the spheroids were fixed in 4% PFA for 1 hour at RT. They were then washed in PBS and added to a bio-wrap (Leica Biosystems Richmond, USA) and placed in a plastic cassette. The cassette was then placed into increasing concentrations (70%, 90% and 100%) of ethanol for 1 hour each, then into histoclear for 1 hour. The fixed spheroids were then placed into paraffin wax at approximately 60 °C for 6 hours to allow the wax to penetrate the spheroids. After 6 hours, the spheroids in wax were placed into a mould and allowed to cool to RT and placed at −20 °C for 1 hour before being sectioned. The sections were added to polysine adhesion glass slides (Epredia, Fisher Scientific, USA) and baked at 60 °C for 2 hours for drying.
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