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Here, we introduce a method to mount fluorescent samples parallel to the optical axis. This mounting allows direct imaging of what would normally be an <jats:italic>x<\/jats:italic>\u2013<jats:italic>z<\/jats:italic> cross\u2010section of the object, in the <jats:italic>x<\/jats:italic>\u2013<jats:italic>y<\/jats:italic> plane of the microscope. With this approach, the <jats:italic>x<\/jats:italic>\u2013<jats:italic>y<\/jats:italic> cross\u2010sections of fluorescent beads were seen to have significantly lower shape\u2010distortions as compared to <jats:italic>x<\/jats:italic>\u2013<jats:italic>z<\/jats:italic> cross\u2010sections reconstructed from confocal <jats:italic>z<\/jats:italic>\u2010stacks. We further tested the method for imaging of nuclear and cellular heights in cultured cells, and found that they are significantly flatter than previously reported. This approach allows improved imaging of the <jats:italic>x<\/jats:italic>\u2013<jats:italic>z<\/jats:italic> cross\u2010section of fluorescent samples.<\/jats:p><\/jats:sec><jats:sec><jats:title>Lay description<\/jats:title><jats:p>Optical distortions are common in confocal microscopy. In particular, the mismatch between the refractive index of the immersion medium of the microscope objective and the refractive index of the sample medium distorts the shapes of fluorescent objects in the <jats:italic>x<\/jats:italic>\u2013<jats:italic>z<\/jats:italic> plane of the microscope. Here, we introduced a method to eliminate the shape\u2010distortion in the <jats:italic>x<\/jats:italic>\u2013<jats:italic>z<\/jats:italic> cross\u2010sections. This was achieved by mounting fluorescent samples on vertical glass slides such that the cross\u2010sections orthogonal to the glass surface could be imaged in the <jats:italic>x<\/jats:italic>\u2013<jats:italic>y<\/jats:italic> plane of the microscope. Our method successfully improved the imaging of nuclear and cellular heights in cultured cells and revealed that the heights were significantly flatter than previously reported with conventional approaches.<\/jats:p><\/jats:sec>","DOI":"10.1111\/jmi.12965","type":"journal-article","created":{"date-parts":[[2020,10,6]],"date-time":"2020-10-06T07:03:50Z","timestamp":1601967830000},"page":"224-230","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["A method for direct imaging of <i>x<\/i>\u2013<i>z<\/i> cross\u2010sections of fluorescent samples"],"prefix":"10.1111","volume":"281","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2888-6320","authenticated-orcid":false,"given":"A.","family":"KATIYAR","sequence":"first","affiliation":[{"name":"Department of Biomedical Engineering Texas A&amp;M University  College Station TX 77840 U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7402-983X","authenticated-orcid":false,"given":"J.D.","family":"ANTANI","sequence":"additional","affiliation":[{"name":"Artie McFerrin Department of Chemical Engineering Texas A&amp;M University  3122 TAMU College Station Texas 77843 U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"B.P.","family":"MCKEE","sequence":"additional","affiliation":[{"name":"Department of Chemical Engineering University of Florida  Gainesville Florida 32611 U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"R.","family":"GUPTA","sequence":"additional","affiliation":[{"name":"Artie McFerrin Department of Chemical Engineering Texas A&amp;M University  3122 TAMU College Station Texas 77843 U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2894-3487","authenticated-orcid":false,"given":"P.P.","family":"LELE","sequence":"additional","affiliation":[{"name":"Artie McFerrin Department of Chemical Engineering Texas A&amp;M University  3122 TAMU College Station Texas 77843 U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"T.P.","family":"LELE","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering Texas A&amp;M University  College Station TX 77840 U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2020,10,19]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1364\/AO.41.000685"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2818.1993.tb03315.x"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2818.2005.01457.x"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2818.2007.01822.x"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41596-020-0360-2"},{"key":"e_1_2_7_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2016.04.047"},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1091\/mbc.E20-01-0073"},{"key":"e_1_2_7_9_1","doi-asserted-by":"crossref","first-page":"690","DOI":"10.1007\/978-0-387-45524-2_39","volume-title":"Photobleaching, in Handbook of Biological Confocal Microscopy","author":"Diaspro A.","year":"2006"},{"key":"e_1_2_7_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bpj.2015.07.006"},{"key":"e_1_2_7_11_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep19689"},{"key":"e_1_2_7_12_1","doi-asserted-by":"publisher","DOI":"10.1002\/jcp.26031"},{"key":"e_1_2_7_13_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep19689"},{"key":"e_1_2_7_14_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep38063"},{"key":"e_1_2_7_15_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41556-017-0030-y"},{"key":"e_1_2_7_16_1","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.201902046"},{"key":"e_1_2_7_17_1","doi-asserted-by":"publisher","DOI":"10.1039\/C9SM01666J"},{"key":"e_1_2_7_18_1","doi-asserted-by":"publisher","DOI":"10.1002\/jcp.28672"},{"key":"e_1_2_7_19_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-05467-x"},{"key":"e_1_2_7_20_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep07362"},{"key":"e_1_2_7_21_1","doi-asserted-by":"publisher","DOI":"10.1007\/s12195-013-0270-2"},{"key":"e_1_2_7_22_1","doi-asserted-by":"publisher","DOI":"10.1038\/nnano.2015.88"},{"issue":"16","key":"e_1_2_7_23_1","first-page":"1744","article-title":"Mechanical stress triggers nuclear remodeling and the formation of transmembrane actin nuclear lines with associated nuclear pore complexes","volume":"31","author":"Hoffman L.M.","journal-title":"Mol. 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