Lyophilization and homogenization of biological samples improves reproducibility and reduces standard deviation in molecular biology techniques.
Freeze drying
Homogenization
Lyophilization
Protein stability
RNA stability
Tissue storage
Journal
Amino acids
ISSN: 1438-2199
Titre abrégé: Amino Acids
Pays: Austria
ID NLM: 9200312
Informations de publication
Date de publication:
Jun 2021
Jun 2021
Historique:
received:
26
08
2020
accepted:
23
04
2021
pubmed:
19
5
2021
medline:
15
12
2021
entrez:
18
5
2021
Statut:
ppublish
Résumé
Lyophilization is a cost-effective method for biological specimen preservation but detailed tissue-specific reference protocols are still lacking. Moreover, data are limited on the long-term stability of proteins and nucleic acids in lyophilized samples.Here, we offer lyophilization protocols for various rat and mouse tissues (kidney, heart, liver, lung, aorta, and skin) coupled with technical hints for optimal sample preparation. We demonstrate that lyophilized samples stored at 4 °C for 20 months can yield protein and RNA of similar quantity and quality to -80 °C storage, while phosphorylated proteins are preserved as well. Freeze-dried and subsequently pulverized samples can provide more consistent, more reliable data especially when investigating focal injuries, such as fibrosis. We developed a protocol for the concentration of biological solutions and achieved 20-times concentration in human peritoneal dialysis effluent solution which enables the previously unattainable detection of proteins in these samples. We established a method for water removal as well as accurate water content measurement of fecal samples, which can be valuable for gut metabolome analysis.Taken together, lyophilization is a valuable tool for the preservation of biological samples with many advantages. We aim to draw attention to the wide range of possibilities offered by freeze drying in pre-clinical or basic research.
Identifiants
pubmed: 34002278
doi: 10.1007/s00726-021-02994-w
pii: 10.1007/s00726-021-02994-w
pmc: PMC8128086
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
917-928Subventions
Organisme : National Research, Development and Innovation Office
ID : FK-124491
Organisme : National Research, Development and Innovation Office
ID : 2017-1.3.1-VKE-2017-00006
Organisme : National Research, Development and Innovation Office
ID : OTKA-PD 131637
Organisme : National Research, Development and Innovation Office
ID : K135398
Organisme : Higher Education Institutional Excellence Program
ID : 2020-4.1.1.-TKP2020-6183069269
Organisme : Higher Education Institutional Excellence Program
ID : 2020-4.1.1.-TKP2020-6183169273
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