Validation of a Novel, Flash-Freezing Method: Aluminum Platform.

Autor: Imrali A; Pancreatic Cancer Research Fund Tissue Bank (PCRFTB) London United Kingdom., Hughes CS; Pancreatic Cancer Research Fund Tissue Bank (PCRFTB) London United Kingdom.; Centre for Tumour Biology, Barts Cancer Institute-A CR-UK Centre of Excellence Queen Mary University of London Charterhouse Square London United Kingdom., Coetzee AS; Centre for Tumour Biology, Barts Cancer Institute-A CR-UK Centre of Excellence Queen Mary University of London Charterhouse Square London United Kingdom., Delvecchio FR; Centre for Tumour Biology, Barts Cancer Institute-A CR-UK Centre of Excellence Queen Mary University of London Charterhouse Square London United Kingdom., Saad A; Pancreatic Cancer Research Fund Tissue Bank (PCRFTB) London United Kingdom., Roberts R; Pancreatic Cancer Research Fund Tissue Bank (PCRFTB) London United Kingdom., Chelala C; Pancreatic Cancer Research Fund Tissue Bank (PCRFTB) London United Kingdom., ChinAleong JA; Barts and the London HPB Centre, Department of Surgery and Pathology, Barts Health NHS Trust The Royal London Hospital London United Kingdom., Kocher HM; Pancreatic Cancer Research Fund Tissue Bank (PCRFTB) London United Kingdom.; Centre for Tumour Biology, Barts Cancer Institute-A CR-UK Centre of Excellence Queen Mary University of London Charterhouse Square London United Kingdom.; Barts and the London HPB Centre, Department of Surgery and Pathology, Barts Health NHS Trust The Royal London Hospital London United Kingdom.
Jazyk: angličtina
Zdroj: Current protocols essential laboratory techniques [Curr Protoc Essent Lab Tech] 2020 Dec; Vol. 21 (1), pp. e46. Date of Electronic Publication: 2020 Nov 25.
DOI: 10.1002/cpet.46
Abstrakt: Stored biological materials should have minimal pre-analytical variations in order to provide researchers with high-quality samples that will give reliable and reproducible results, yet methods of storage should be easy to implement, with minimal cost and health hazard. Frozen tissue samples are a valuable biological resource. Here we compare different methods, such as liquid nitrogen (LN) or dry ice (DI), to a cheap and safe alternative using an aluminum platform (AP). Murine fresh liver and pancreas tissues were used with varying lengths of warm ischemia time. Quality assessment was based on histological evaluation, DNA and RNA extraction and quantification, and RNA degradation analysis, as well preservation of antigens for immunofluorescence, in a blinded manner. Both in superficial and deep tissue sections, based on histological assessment, AP is superior to DI, or as good as LN techniques in terms of presence of ice crystals, cutting artifacts, and overall quality/structural preservation. DNA and RNA were successfully extracted in reasonable quantities from all freezing techniques, but RNA degradation was seen for pancreas samples across all techniques. Immunofluorescence with cytokeratin8 (CK-8), alpha smooth muscle actin (αSMA), CD3, and B220 shows equally good outcomes for AP and LN, which are better than DI. The aluminum platform is a cheap, yet reliable method to freeze samples, rapidly preserving histological, antigenic, and DNA/RNA quality. Wider testing is required across different sample types. © 2020 The Authors. Basic Protocol : Flash-freezing fresh tissue with aluminum platform Alternate Protocol 1 : Freezing fresh tissue with liquid nitrogen Alternate Protocol 2 : Freezing fresh tissue with dry ice.
Competing Interests: The authors declare that there is no conflict of interest.
(© 2020 The Authors.)
Databáze: MEDLINE
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