{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,31]],"date-time":"2026-10-31T08:30:11Z","timestamp":1793435411444,"version":"4.5.0"},"reference-count":67,"publisher":"Radiation Research Society","issue":"1","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Radiation Research"],"published-print":{"date-parts":[[2022,4,7]]},"abstract":"<jats:p>Clonogenic assays are the gold standard for measuring cell clonogenic survival and enable quantification of a cell line's radiosensitivity through the calculation of the surviving fraction, the ratio of cell clusters (colonies) formed after radiation exposure compared to the number formed without exposure. Such studies regularly utilize Cs-137 irradiators. While uncertainties for specific procedural aspects have been described previously, a comprehensive review has not been completed. We therefore quantified uncertainties associated with clonogenic assays performed using a Cs-137 Shepherd irradiator, and a recently established brachytherapy afterloader in vitro radiation delivery apparatus (BAIRDA), through a series of experiments and a literature review. The clonogenic assay is subject to uncertainties that affect the determination of the surviving fraction (e.g., accuracy of the number of cells seeded, potential effects of hypothermia, and the threshold number of cells for a cluster to be identified as a colony). Furthermore, dose delivery uncertainties related to both the Cs-137 irradiator and BAIRDA were also quantified. The combined standard (k = 1) uncertainty was \u00b1 6.0% in the surviving fraction for the Cs-137 irradiator (\u00b16.3% for BAIRDA), up to \u00b1 2.2% in the dose delivered by the Cs-137 irradiator, and up to \u00b1 4.3% in the dose delivered by BAIRDA. The largest individual uncertainties were associated with the number of cells seeded on a plate (3.4%) and inter-observer variability in counting (4.1%), suggesting that effective reduction of uncertainties in the conduct of the clonogenic assay may provide the greatest relief on the uncertainty budget. Finally, measurable impact on experimental findings was assessed by applying this uncertainty to clonogenic assays of SW756 cells using either a Cs-137 irradiator or BAIRDA, introducing a maximum shift in the reported radiobiological parameters \u03b1\/\u03b2 and T1\/2 of 0.3 Gy and 0.4 h, respectively, while the 95% confidence interval increased by 0.5 Gy and decreased by 0.4 h, respectively. Though the overall impact on radiobiological parameter estimation was small, the individual uncertainties could have a significant influence in other applications of in vitro experiments in radiation biology. Hence, better understanding of the uncertainties associated with both clonogenic assays and the radiation source used can improve the accuracy of experimental analysis and reproducibility of the results.<\/jats:p>","DOI":"10.1667\/rade-21-00205.1","type":"journal-article","created":{"date-parts":[[2022,4,7]],"date-time":"2022-04-07T17:10:05Z","timestamp":1649351405000},"page":"40-56","source":"Crossref","is-referenced-by-count":0,"title":["Uncertainties Associated with Clonogenic Assays using a Cs-137 Irradiator and Ir-192 Afterloader: A Comprehensive Compilation for Radiation Researchers"],"prefix":"10.1667","volume":"198","author":[{"given":"Braden","family":"Chow","sequence":"first","affiliation":[{"name":"Department of Oncology, University of Alberta, Edmonton, Alberta, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brad","family":"Warkentin","sequence":"additional","affiliation":[{"name":"Department of Oncology, University of Alberta, Edmonton, Alberta, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Malcolm","family":"McEwen","sequence":"additional","affiliation":[{"name":"Ionizing Radiation Standards, National Research Council of Canada, Ottawa, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fleur","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Oncology, University of Alberta, Edmonton, Alberta, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kareena","family":"Nanda","sequence":"additional","affiliation":[{"name":"Department of Oncology, University of Alberta, Edmonton, Alberta, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Armin M","family":"Gamper","sequence":"additional","affiliation":[{"name":"Department of Oncology, University of Alberta, Edmonton, Alberta, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Geetha","family":"Menon","sequence":"additional","affiliation":[{"name":"Department of Oncology, University of Alberta, Edmonton, Alberta, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1243","reference":[{"key":"5","doi-asserted-by":"crossref","unstructured":"NAP Franken, HM Rodermond, J Stap, et al.  \"Nat Protoc.\"  1, \"Clonogenic assay of cells in vitro.\" 2315 (2006)","DOI":"10.1038\/nprot.2006.339"},{"key":"23","doi-asserted-by":"crossref","unstructured":"DJ. 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American Association of Physicists in Medicine.\" 2093 (1998)","DOI":"10.1118\/1.598407"},{"key":"38","doi-asserted-by":"crossref","unstructured":"A Niroomand-Rad, S Chiu-Tsao, MP Grams, et al.  \"Med Phys.\"  47, \"Report of AAPM Task Group 235 Radiochromic Film Dosimetry: An Update to TG-55.\" 5986 (2020)","DOI":"10.1002\/mp.14497"},{"key":"39","doi-asserted-by":"crossref","unstructured":"H Morrison, G Menon, RS Sloboda,  \"Med Phys.\"  41, \"Radiochromic film calibration for low-energy seed brachytherapy dose measurement.\" 072101 (2014)","DOI":"10.1118\/1.4881146"},{"key":"41","doi-asserted-by":"crossref","unstructured":"M Dos Santos, V Paget, MB Kacem, et al.  \"Int J Radiat Biol.\"  94, \"Importance of dosimetry protocol for cell irradiation on a low x-rays facility and consequences for the biological response.\" 597 (2018)","DOI":"10.1080\/09553002.2018.1466205"},{"key":"42","doi-asserted-by":"crossref","unstructured":"S Strohmaier  and G Zwierzchowski  \"J Contemp Brachytherapy.\"  3, \"Comparison of 60Co and 192I sources in HDR brachytherapy.\" 199 (2011)","DOI":"10.5114\/jcb.2011.26471"},{"key":"43","doi-asserted-by":"crossref","unstructured":"LA DeWerd, GS Ibbott, AS Meigooni, et al.  \"Med Phys.\"  38, \"A dosimetric uncertainty analysis for photon-emitting brachytherapy sources: Report of AAPM Task Group No. 138 and GEC-ESTRO.\" 782 (2011)","DOI":"10.1118\/1.3533720"},{"key":"44","doi-asserted-by":"crossref","unstructured":"K Tanderup, D Georg, R Potter, et al.  \"Semin Radiat Oncol.\"  20, \"Adaptive management of cervical cancer radiotherapy.\" 121 (2010)","DOI":"10.1016\/j.semradonc.2009.11.006"},{"key":"45","doi-asserted-by":"crossref","unstructured":"C Kirisits, MJ Rivard, D Baltas, et al.  \"Radiother Oncol.\"  110, \"Review of clinical brachytherapy uncertainties: Analysis guidelines of GEC-ESTRO and the AAPM.\" 199 (2014)","DOI":"10.1016\/j.radonc.2013.11.002"},{"key":"46","doi-asserted-by":"crossref","unstructured":"GV Menon, MC Carlone, RS Sloboda,  \"Phys Med Biol.\"  53, \"Transit dose contributions to intracavitary and interstitial PDR brachytherapy treatments.\" 3447 (2008)","DOI":"10.1088\/0031-9155\/53\/13\/003"},{"key":"47","unstructured":"R van der Laarse, Y Niatsetki, J Henning, et al.  \"Flexitron\u00ae and microSelectron\u00ae HDR\/PDR afterloaders' transit time and dose.\"  12 (2018)"},{"key":"48","doi-asserted-by":"crossref","unstructured":"MJ Rivard, BM Cousey, LA DeWerd, et al.  \"Med Phys.\"  31, \"Update of AAPM Task Group No. 43 Report: A revised AAPM protocol for brachytherapy dose calculations.\" 633 (2004)","DOI":"10.1118\/1.1646040"},{"key":"49","doi-asserted-by":"crossref","unstructured":"SA White, G Landry, GP Fonseca, et al.  \"Med Phys.\"  41, \"Comparison of TG-43 and TG-186 in breast irradiation using a low energy electronic brachytherapy source.\" 061701 (2014)","DOI":"10.1118\/1.4873319"},{"key":"50","doi-asserted-by":"crossref","unstructured":"MJ Rivard, JLM Venselaar, L Beaulieu,  \"Med Phys.\"  36, \"The evolution of brachytherapy treatment planning.\" 2136 (2009)","DOI":"10.1118\/1.3125136"},{"key":"51","doi-asserted-by":"crossref","unstructured":"L Beaulieu, A Carlsson Tedgren, J Carrier, et al.  \"Med Phys.\"  39, \"Report of the Task Group 186 on model-based dose calculation methods in brachytherapy beyond the TG-43 formalism: current status and recommendations for clinical implementation.\" 6208 (2012)","DOI":"10.1118\/1.4747264"},{"key":"52","doi-asserted-by":"crossref","unstructured":"J Hofbauer, C Kirisits, A Resch, et al.  \"J Contemp Brachytherapy.\"  8, \"Impact of heterogeneity-corrected dose calculation using a grid-based Boltzmann solver on breast and cervix cancer brachytherapy.\" 143 (2016)","DOI":"10.5114\/jcb.2016.59352"},{"key":"53","doi-asserted-by":"crossref","unstructured":"EP Pappas, V Peppa, CJ Hourdakis, et al.  \"Phys Medica.\"  45, \"On the use of a novel Ferrous Xylenol-orange gelatin dosimeter for HDR brachytherapy commissioning and quality assurance testing.\" 162 (2018)","DOI":"10.1016\/j.ejmp.2017.12.015"},{"key":"54","unstructured":"\"Evaluation of measurement data - Guide to the expression of uncertainty in measurement.\"  (2008)"},{"key":"55","doi-asserted-by":"crossref","unstructured":"JD Chapman  and AE Nahum  \"Radiotherapy treatment planning.\"  (2016)","DOI":"10.1201\/b18349"},{"key":"56","doi-asserted-by":"crossref","unstructured":"J. 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