BACKGROUND: Fresh frozen tissue from radical prostatectomy specimens is highly valuable material for research on gene expression and cellular metabolites. The purpose of this study was to develop a standardized method to provide a representative high quality research sample from radical prostatectomy specimens without interfering with the routine histopathological procedure. METHODS: A complete transversal slice is collected and snap-frozen before formalin fixation and routine processing of the remaining gland. The freezing preserves the original geometric shape, thus allowing subsampling of specific cell populations without thawing. RNA was extracted from 53 cylindrical subsamples (diameter 3 mm, thickness 2 mm) from 16 consecutive frozen slices. The histological pattern was evaluated by microscopy of a cryosection from sample before further analysis. RESULTS: Using this novel harvesting method close to 400 slices have been collected. Whenever tumor was present in both adjacent surrounding hematoxylin-eosin sections, we found cancer in 88% of the frozen slices. The extracted RNA showed very high quality with a mean RNA integrity number of 9.16 (SD 0.53). The MR spectra showed metabolic profiles containing several resonances, which deserve further evaluation as possible biomarkers for prostate cancer. After MR analysis the RNA was still highly intact with a mean RNA integrity number of 8.40 (SD 1.53), which makes it possible to correlate transcriptomic and metabolomic profiles of the extracted samples. CONCLUSION: We present a safe and standardized method for procurement of a high quality fresh frozen prostate slice, suitable for gene expression analysis and MR spectroscopy.
BACKGROUND: Fresh frozen tissue from radical prostatectomy specimens is highly valuable material for research on gene expression and cellular metabolites. The purpose of this study was to develop a standardized method to provide a representative high quality research sample from radical prostatectomy specimens without interfering with the routine histopathological procedure. METHODS: A complete transversal slice is collected and snap-frozen before formalin fixation and routine processing of the remaining gland. The freezing preserves the original geometric shape, thus allowing subsampling of specific cell populations without thawing. RNA was extracted from 53 cylindrical subsamples (diameter 3 mm, thickness 2 mm) from 16 consecutive frozen slices. The histological pattern was evaluated by microscopy of a cryosection from sample before further analysis. RESULTS: Using this novel harvesting method close to 400 slices have been collected. Whenever tumor was present in both adjacent surrounding hematoxylin-eosin sections, we found cancer in 88% of the frozen slices. The extracted RNA showed very high quality with a mean RNA integrity number of 9.16 (SD 0.53). The MR spectra showed metabolic profiles containing several resonances, which deserve further evaluation as possible biomarkers for prostate cancer. After MR analysis the RNA was still highly intact with a mean RNA integrity number of 8.40 (SD 1.53), which makes it possible to correlate transcriptomic and metabolomic profiles of the extracted samples. CONCLUSION: We present a safe and standardized method for procurement of a high quality fresh frozen prostate slice, suitable for gene expression analysis and MR spectroscopy.
Authors: Rebecca Buchholz; Sebastian Krossa; Maria K Andersen; Michael Holtkamp; Michael Sperling; Uwe Karst; May-Britt Tessem Journal: Metallomics Date: 2022-03-28 Impact factor: 4.526
Authors: Guro F Giskeødegård; Helena Bertilsson; Kirsten M Selnæs; Alan J Wright; Tone F Bathen; Trond Viset; Jostein Halgunset; Anders Angelsen; Ingrid S Gribbestad; May-Britt Tessem Journal: PLoS One Date: 2013-04-23 Impact factor: 3.240
Authors: Therese S Høiem; Maria K Andersen; Marta Martin-Lorenzo; Rémi Longuespée; Britt S R Claes; Anna Nordborg; Frédéric Dewez; Benjamin Balluff; Marco Giampà; Animesh Sharma; Lars Hagen; Ron M A Heeren; Tone F Bathen; Guro F Giskeødegård; Sebastian Krossa; May-Britt Tessem Journal: Proteomics Date: 2022-03-03 Impact factor: 5.393
Authors: Morten Beck Rye; Helena Bertilsson; Finn Drabløs; Anders Angelsen; Tone F Bathen; May-Britt Tessem Journal: BMC Med Genomics Date: 2014-08-12 Impact factor: 3.063
Authors: May-Britt Tessem; Helena Bertilsson; Anders Angelsen; Tone F Bathen; Finn Drabløs; Morten Beck Rye Journal: PLoS One Date: 2016-04-21 Impact factor: 3.240