{"id":3388,"date":"2020-05-29T10:35:58","date_gmt":"2020-05-29T14:35:58","guid":{"rendered":"https:\/\/engineering.jhu.edu\/lefd\/?page_id=3388"},"modified":"2020-06-02T11:30:32","modified_gmt":"2020-06-02T15:30:32","slug":"lung-cell","status":"publish","type":"page","link":"https:\/\/engineering.jhu.edu\/lefd\/lung-cell\/","title":{"rendered":"Lung Cell Exposure Chamber"},"content":{"rendered":"<h3>People<\/h3>\n<table style=\"height: auto;\" width=\"700\" align=\"center\">\n<tbody>\n<tr>\n<td width=\"250\">Postdoc<\/td>\n<td width=\"250\">Lakshmana Chandrala<\/td>\n<\/tr>\n<tr>\n<td width=\"250\">Graduate Student<\/td>\n<td width=\"250\">Name<\/td>\n<\/tr>\n<tr>\n<td width=\"250\">Project Supervisor<\/td>\n<td width=\"250\">Prof. Joseph Katz<\/td>\n<\/tr>\n<tr>\n<td width=\"250\">Design &amp; Technical Support<\/td>\n<td width=\"250\">Dr. Yury Ronzhes<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<h3>Introduction<\/h3>\n<p style=\"text-align: justify;\">Aerosolization of oil by wave splashing on crude oil slicks may pose a respiratory health threat to oil-spill workers and communities nearby. However, the health effects of inhaling oily aerosols remain mostly unknown. Exposure systems for <em>in vitro\u00a0<\/em> testing of bronchial epithelial cells to airborne environmental agents are crucial in understanding the airway disease pathogenesis. Several such systems have been developed and standardized over the years for toxicity testing of airborne particles. However, a significant limitation of these systems is that they do not permit direct observation on the cells during the exposure and the cells have to be removed from the chamber for microscopic observations for, e.g., Ciliary Beat Frequency (CBF) measurements.\u00a0 Hence, despite considerable efforts, little is known about the biological responses of epithelial cells to airborne particles during the exposure.<\/p>\n<h3>Objective<\/h3>\n<p style=\"text-align: justify;\">The present study introduces an exposure chamber for Real-Time Examination of Cell Exposure (RTECE) that enables <em>in-situ<\/em> live imaging of cell cultures while being exposed to airborne materials under controlled conditions.<\/p>\n<h3>Real-Time Examination of Cell Exposure system<\/h3>\n<h3><a href=\"https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LK123.png\"><img loading=\"lazy\" decoding=\"async\" class=\"size-full wp-image-3404 aligncenter\" src=\"https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LK123.png\" alt=\"\" width=\"1080\" height=\"507\" srcset=\"https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LK123.png 1080w, https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LK123-300x141.png 300w, https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LK123-1024x481.png 1024w, https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LK123-200x94.png 200w, https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LK123-768x361.png 768w, https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LK123-150x70.png 150w\" sizes=\"auto, (max-width: 1080px) 100vw, 1080px\" \/><\/a>Features of the system<\/h3>\n<ul>\n<li>In-situ imaging of cells involving different illumination and microscopic observation techniques<\/li>\n<li>Optimized nozzle for uniform deposition of aerosols on the cell culture surface<\/li>\n<li>Disposable 3D-printed nozzles to avoid cross-contamination between the experiments.<\/li>\n<\/ul>\n<h3 style=\"text-align: justify;\"><a href=\"https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_components-of-chamber.png\"><img loading=\"lazy\" decoding=\"async\" class=\" wp-image-3407 aligncenter\" src=\"https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_components-of-chamber.png\" alt=\"\" width=\"482\" height=\"405\" srcset=\"https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_components-of-chamber.png 1276w, https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_components-of-chamber-300x252.png 300w, https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_components-of-chamber-1024x861.png 1024w, https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_components-of-chamber-200x168.png 200w, https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_components-of-chamber-768x646.png 768w, https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_components-of-chamber-150x126.png 150w\" sizes=\"auto, (max-width: 482px) 100vw, 482px\" \/><\/a>Optical Modalities<\/h3>\n<p style=\"text-align: justify;\">Two optical modalities, oblique illumination microscopy and fluorescence imaging, are employed to visualize the cell culture <em>in situ<\/em>. Oblique illumination microscopy is used for monitoring the cilia motion and cell migration, and fluorescence microscopy is used for observing fluorescently-stained cells.<\/p>\n<h3><a href=\"https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_optical_modalities.png\"><img loading=\"lazy\" decoding=\"async\" class=\"size-full wp-image-3413 aligncenter\" src=\"https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_optical_modalities.png\" alt=\"\" width=\"2337\" height=\"1219\" srcset=\"https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_optical_modalities.png 2337w, https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_optical_modalities-300x156.png 300w, https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_optical_modalities-1024x534.png 1024w, https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_optical_modalities-200x104.png 200w, https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_optical_modalities-768x401.png 768w, https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_optical_modalities-1536x801.png 1536w, https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_optical_modalities-2048x1068.png 2048w, https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_optical_modalities-150x78.png 150w\" sizes=\"auto, (max-width: 2337px) 100vw, 2337px\" \/><\/a>Results<\/h3>\n<p style=\"text-align: justify;\">To demonstrate the capability of this system, a confluent monolayer of Normal Human Bronchial Epithelial (NHBE) cells are subjected to two exposure separated by a rest period of 60 min, each consisting of two cigarettes.<\/p>\n<p style=\"text-align: center;\"><a href=\"https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_cilia_movie.gif\"><img loading=\"lazy\" decoding=\"async\" class=\"size-full wp-image-3416 aligncenter\" src=\"https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_cilia_movie.gif\" alt=\"\" width=\"728\" height=\"676\" \/><\/a>Sample movie showing the time evolution of cilia during the exposure to cigarette smoke. The video is obtained after the high-pass filtering the time-series of the intensity of each pixel.<\/p>\n<h3>In situ measurement of CBF:<\/h3>\n<ul>\n<li style=\"text-align: justify;\">NHBE cells are subjected to two exposures, each exposure consisting of two cigarettes.<\/li>\n<li style=\"text-align: justify;\">Using oblique illumination microscopy, the RTECE is used for in situ measurements of the time evolution of CBF.<\/li>\n<\/ul>\n<h3><a href=\"https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_time_cbf.png\"><img loading=\"lazy\" decoding=\"async\" class=\"size-full wp-image-3421 aligncenter\" src=\"https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_time_cbf.png\" alt=\"\" width=\"1982\" height=\"887\" srcset=\"https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_time_cbf.png 1982w, https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_time_cbf-300x134.png 300w, https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_time_cbf-1024x458.png 1024w, https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_time_cbf-200x90.png 200w, https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_time_cbf-768x344.png 768w, https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_time_cbf-1536x687.png 1536w, https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_time_cbf-150x67.png 150w\" sizes=\"auto, (max-width: 1982px) 100vw, 1982px\" \/><\/a>In situ measurement of Cell migration:<\/h3>\n<ul>\n<li style=\"text-align: justify;\">Using Particle image velocimetry, the cell motions are monitored for twelve hours.<\/li>\n<\/ul>\n<h3>Exposure to smoke increases the spatially-averaged cell velocity by an order of magnitude<br \/>\n<a href=\"https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_time_migration.png\"><img loading=\"lazy\" decoding=\"async\" class=\"size-full wp-image-3423 aligncenter\" src=\"https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_time_migration.png\" alt=\"\" width=\"2032\" height=\"1370\" srcset=\"https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_time_migration.png 2032w, https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_time_migration-300x202.png 300w, https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_time_migration-1024x690.png 1024w, https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_time_migration-200x135.png 200w, https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_time_migration-768x518.png 768w, https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_time_migration-1536x1036.png 1536w, https:\/\/engineering.jhu.edu\/lefd\/wp-content\/uploads\/2020\/05\/LC_time_migration-150x101.png 150w\" sizes=\"auto, (max-width: 2032px) 100vw, 2032px\" \/><\/a>Publications<\/h3>\n<p style=\"text-align: justify;\">Chandrala, L. D., Afshar-Mohajer, N., Nishida, K., Ronzhes, Y., Sidhaye, V. K., Koehler, K., &amp; Katz, J. (2019). <a href=\"https:\/\/www.nature.com\/articles\/s41598-019-43784-5\" target=\"_blank\" rel=\"noopener noreferrer\"><em>A Device for measuring the in-situ response of Human Bronchial Epithelial Cells to airborne environmental agents<\/em><\/a>. Scientific reports, 9(1), 1-12.<\/p>\n","protected":false},"excerpt":{"rendered":"<p>People Postdoc Lakshmana Chandrala Graduate Student Name Project Supervisor Prof. Joseph Katz Design &amp; Technical Support Dr. Yury Ronzhes Introduction Aerosolization of oil by wave splashing on crude oil slicks may pose a respiratory health threat to oil-spill workers and &hellip; <a href=\"https:\/\/engineering.jhu.edu\/lefd\/lung-cell\/\">Continue reading <span class=\"meta-nav\">&rarr;<\/span><\/a><\/p>\n","protected":false},"author":5558,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"_exactmetrics_skip_tracking":false,"_exactmetrics_sitenote_active":false,"_exactmetrics_sitenote_note":"","_exactmetrics_sitenote_category":0,"footnotes":""},"class_list":["post-3388","page","type-page","status-publish","hentry"],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v27.8 - https:\/\/yoast.com\/product\/yoast-seo-wordpress\/ -->\n<title>Lung Cell Exposure Chamber - Laboratory for Experimental Fluid Dynamics<\/title>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/engineering.jhu.edu\/lefd\/lung-cell\/\" \/>\n<meta property=\"og:locale\" content=\"en_US\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Lung Cell Exposure Chamber - Laboratory for Experimental Fluid Dynamics\" \/>\n<meta property=\"og:description\" content=\"People Postdoc Lakshmana Chandrala Graduate Student Name Project Supervisor Prof. Joseph Katz Design &amp; 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