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Over the years, extensive research has focused on both the structure and function of ribosomes. However, a fundamental question \u2013 how many ribosomes are present within whole cells \u2013 has remained largely unaddressed. In this study, we developed a microscopic method to quantify the total number of ribosomes in hTERT\u2010RPE\u20101 cells and in nematode cells from various tissues of <jats:italic>Caenorhabditis elegans<\/jats:italic> hermaphrodites. Using electron tomography of high\u2010pressure frozen, freeze\u2010substituted and resin\u2010embedded samples, we determined that the ribosome number in hTERT\u2010RPE\u20101 cells is in the same order of magnitude as biochemical measurements obtained via RNA capillary electrophoresis. As expected, control worms exhibited a higher number of ribosomes compared to RNA polymerase I A subunit (RPOA\u20101)\u2010depleted worms in two out of three analysed tissue types. Our imaging\u2010based approach complements established biochemical methods by enabling direct quantification of ribosome numbers in specific samples. This method offers a powerful tool for advancing our understanding of ribosome localisation and distribution in cells and tissues across diverse model systems.<\/jats:p>","DOI":"10.1111\/jmi.13380","type":"journal-article","created":{"date-parts":[[2025,1,15]],"date-time":"2025-01-15T14:49:14Z","timestamp":1736952554000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["In situ quantification of ribosome number by electron tomography"],"prefix":"10.1111","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-0812-383X","authenticated-orcid":false,"given":"Mounir","family":"El Hankouri","sequence":"first","affiliation":[{"name":"Faculty of Medicine Carl Gustav Carus Experimental Center Technische Universit\u00e4t Dresden  Dresden Germany"},{"name":"Cluster of Excellence Physics of Life Technische Universit\u00e4t Dresden  Dresden Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marco","family":"Nousch","sequence":"additional","affiliation":[{"name":"Department of Developmental Genetics Institute of Biology Martin\u2010Luther\u2010Universit\u00e4t Halle\u2010Wittenberg  Halle Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8030-8851","authenticated-orcid":false,"given":"Aayush","family":"Poddar","sequence":"additional","affiliation":[{"name":"Faculty of Medicine Carl Gustav Carus Experimental Center Technische Universit\u00e4t Dresden  Dresden Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0203-1436","authenticated-orcid":false,"given":"Thomas","family":"M\u00fcller\u2010Reichert","sequence":"additional","affiliation":[{"name":"Faculty of Medicine Carl Gustav Carus Experimental Center Technische Universit\u00e4t Dresden  Dresden Germany"},{"name":"Faculty of Medicine Carl Gustav Carus Core Facility Cellular Imaging Technische Universit\u00e4t Dresden  Dresden Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3017-0978","authenticated-orcid":false,"given":"Gunar","family":"Fabig","sequence":"additional","affiliation":[{"name":"Faculty of Medicine Carl Gustav Carus Experimental Center Technische Universit\u00e4t Dresden  Dresden Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2025,1,15]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.59.3.777"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.32607\/actanaturae.11541"},{"key":"e_1_2_6_4_1","doi-asserted-by":"publisher","DOI":"10.32607\/actanaturae.11540"},{"key":"e_1_2_6_5_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1212642"},{"key":"e_1_2_6_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ab.2007.09.032"},{"key":"e_1_2_6_7_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature02046"},{"key":"e_1_2_6_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature04785"},{"key":"e_1_2_6_9_1","doi-asserted-by":"publisher","DOI":"10.1002\/pmic.202000039"},{"key":"e_1_2_6_10_1","doi-asserted-by":"publisher","DOI":"10.1093\/genetics\/77.1.71"},{"key":"e_1_2_6_11_1","doi-asserted-by":"publisher","DOI":"10.1101\/pdb.prot072348"},{"key":"e_1_2_6_12_1","doi-asserted-by":"publisher","DOI":"10.1038\/35888"},{"key":"e_1_2_6_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1046\u20102023(03)00050\u20101"},{"key":"e_1_2_6_14_1","doi-asserted-by":"publisher","DOI":"10.21769\/BioProtoc.4849"},{"key":"e_1_2_6_15_1","first-page":"151","volume-title":"Methods in cell biology","author":"Kiewisz R.","year":"2021"},{"key":"e_1_2_6_16_1","first-page":"119","volume-title":"Methods in cell biology","author":"Fabig G.","year":"2019"},{"key":"e_1_2_6_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/0011\u20102240(74)90007\u20108"},{"key":"e_1_2_6_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jsb.2020.107492"},{"key":"e_1_2_6_19_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365\u20102818.2003.01250.x"},{"key":"e_1_2_6_20_1","doi-asserted-by":"publisher","DOI":"10.1017\/S1431927603445911"},{"key":"e_1_2_6_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jsb.2005.07.007"},{"key":"e_1_2_6_22_1","doi-asserted-by":"publisher","DOI":"10.1006\/jsbi.1997.3919"},{"key":"e_1_2_6_23_1","doi-asserted-by":"publisher","DOI":"10.1006\/jsbi.1996.0013"},{"key":"e_1_2_6_24_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jsb.2016.07.011"},{"key":"e_1_2_6_25_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365\u20102818.1977.tb00012.x"},{"key":"e_1_2_6_26_1","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1007\/978-0-387-69008-7_2","volume-title":"Electron tomography: Methods for three\u2010dimensional visualization of structures in the cell","author":"Luther P. 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