A Method of Low-Temperature Storing of Agarose Slides with Lysed Cells

• Published in: Bulletin of experimental biology and medicine 2013-10, Vol.155 (6), p.757-759
• Main Authors: Kuznetsova, E. A., Dyukina, A. R., Chernigina, I. A., Sirota, N. P.
• Format: Article
• Language: English
• Subjects: Adult; Aged; Animals; Biomedical and Life Sciences; Biomedicine; Blood Banks; Cell Biology; Cell Extracts; Cryopreservation; DNA; DNA Damage; Female; Glycerin; Glycerol; Humans; Internal Medicine; Laboratory Medicine; Leukocytes, Mononuclear - physiology; Male; Methods; Mice; Middle Aged; Pathology; Phosphates; Sepharose - chemistry; Tissue Embedding; Young Adult
• ISSN: 0007-4888; 1573-8221
• DOI: 10.1007/s10517-013-2245-7

A method has been developed for a long-term low-temperature storage (-10 to -15°C) of the agarose slides with nucleoids (lysed eukaryotic cells). After lysis of agarose-immobilized cells, the slides were incubated for 30 min in phosphate buffer with 50% glycerol and 100 mM EDTA, thereupon they were stored in a freezer at -10 to -15°C. After long-term storage, the slides were re-incubated for 30 min in lysing solution. The measurements of the baseline and in vitro induced DNA damage in nucleoids of the human and mouse leukocytes, which had been stored in agarose slides at low temperature, showed that DNA damage level determined after a 40-day storage did not significantly differ from that of the fresh slides. The advanced storage method is simple and reliable; it opens the way to avoid cryopreservation of the biological samples and to process little by little a great number of the identically prepared slides.