{"id":11869,"date":"2017-12-08T11:52:48","date_gmt":"2017-12-08T10:52:48","guid":{"rendered":"https:\/\/news.embl.de\/?p=11869"},"modified":"2024-03-22T11:51:52","modified_gmt":"2024-03-22T10:51:52","slug":"alasdair-mcdowall-slow-flash-freezing","status":"publish","type":"post","link":"https:\/\/www.embl.org\/news\/alumni\/alasdair-mcdowall-slow-flash-freezing\/","title":{"rendered":"Alasdair McDowall\u2019s slow road to flash freezing"},"content":{"rendered":"\n<p>In the late 1970s, water was the bane of the microscopist\u2019s existence. Since water doesn\u2019t stay put under the powerful vacuum of an electron microscope, trying to image a live cell\u2014plump with water\u2014resulted in a vacuum-dried, shriveled mass. The best option at the time was to dehydrate a cell in advance and fix it with chemicals and resins to preserve some semblance of its lifelike shape. But there was one other tantalising option to image a cell that didn\u2019t require drying it out: lock the cell\u2019s water in place by freezing it.<\/p>\n\n\n\n<p>Still in its infancy, this solution also posed its own problems. In order to use an electron microscope to see a frozen cell from, say, a thin slice of rat kidney or human liver meant trying to make out its critical structures\u2014swirls of cytoskeleton strands, rippled hillocks of reticulum\u2014despite the black spots of ice crystals that the electrons in the microscope can\u2019t penetrate. In the worst case, when the frozen water turned particularly vicious, ice crystals would spear the cell from the inside, causing it to burst.<\/p>\n\n\n\n<div class=\"wp-block-image wp-image-11872 size-full vf-u-width__30\"><figure class=\"vf-figure  | vf-figure--align vf-figure--align-inline-start  \"><img loading=\"lazy\" decoding=\"async\" width=\"310\" height=\"442\" class=\"vf-figure__image\" src=\"https:\/\/news.embl.de\/wp-content\/uploads\/2017\/12\/171208-mcdowall2.jpg\" alt=\"Alasdair McDowall at work at EMBL\" class=\"wp-image-11872\" srcset=\"https:\/\/www.embl.org\/news\/wp-content\/uploads\/2017\/12\/171208-mcdowall2.jpg 310w, https:\/\/www.embl.org\/news\/wp-content\/uploads\/2017\/12\/171208-mcdowall2-210x300.jpg 210w\" sizes=\"auto, (max-width: 310px) 100vw, 310px\" \/><figcaption class=\"vf-figure__caption\">Alasdair McDowall at work at EMBL. PHOTO: Alasdair McDowall<\/figcaption><\/figure><\/div>\n\n\n\n<p>In 1978, the dream of EMBL group leader Jacques Dubochet was to freeze water so fast that crystals couldn\u2019t form. The water would become vitrified\u2014glasslike\u2014allowing researchers to see cells as they had been in their living state using an electron microscope. For <a href=\"http:\/\/beckmaninstitute.caltech.edu\/tem.shtml\" target=\"_blank\" rel=\"noopener noreferrer\">Alasdair McDowall<\/a>, a Scot and self-described nomad who worked as Dubochet\u2019s research technician, this project was a departure from the type of histopathology work he\u2019d done before.<\/p>\n\n\n\n<p>\u201cI was a good at cutting thin sections of biological samples and hadn\u2019t thought a lot about cryobiology before,\u201d says McDowall. \u201cBut I wasn\u2019t at EMBL long before I got the bug, too,\u201d<\/p>\n\n\n\n<p>The path to vitrification, though, was uncertain. Initially, Dubochet\u2019s group thought the speed at which they cooled the sample would be important. McDowall helped to rig up a device that shot samples into a frigid bath of liquid nitrogen.<\/p>\n\n\n\n<p>They also tried minimising the size of the sample, reasoning that a smaller sample might freeze fast enough to beat crystal formation. After trying to vitrify tiny droplets, they figured even those might be too big, so they tried spraying the sample onto a grid in order to get an ultrathin layer of cells. They experimented with using different cryogens\u2014 liquid nitrogen slush, freons and propanes, ethanes and all sorts of ethylenes\u2014 and cryoprotectants to shield the cells from damaging ice crystals.<\/p>\n\n\n\n<div class=\"wp-block-image wp-image-11876 size-full\"><figure class=\"vf-figure  | vf-figure--align vf-figure--align-centered \"><img loading=\"lazy\" decoding=\"async\" width=\"620\" height=\"300\" class=\"vf-figure__image\" src=\"https:\/\/news.embl.de\/wp-content\/uploads\/2017\/12\/171208-mcdowall3.jpeg\" alt=\"The original cryoholder developed by Dubochet, McDowall, and their team\" class=\"wp-image-11876\" srcset=\"https:\/\/www.embl.org\/news\/wp-content\/uploads\/2017\/12\/171208-mcdowall3.jpeg 620w, https:\/\/www.embl.org\/news\/wp-content\/uploads\/2017\/12\/171208-mcdowall3-300x145.jpeg 300w\" sizes=\"auto, (max-width: 620px) 100vw, 620px\" \/><figcaption class=\"vf-figure__caption\">The original cryoholder developed by Dubochet, McDowall, and their team. PHOTO: EMBL<\/figcaption><\/figure><\/div>\n\n\n\n<p>After each shift in technique, McDowall peered at the latest sample in a microscope, hoping to see an improvement. \u201cMainly, what we saw was useless,\u201d he says, smiling.<\/p>\n\n\n\n<p>Then one afternoon in 1980, after plunging a sample into ethane, a cryogen they hadn\u2019t yet tried, Alasdair McDowall peered at a grid under a microscope and saw something strange: diffuse, round drops. He called over his lab head, Jacques Dubochet, to have a look. But Dubochet hadn\u2019t seen the structureless splotches before either, so he bombarded the sample with electrons to determine whether or not it had a crystal structure using a process called diffraction. In the image produced by the diffraction, he expected to see a regular pattern of concentric circles, indicating the substance was crystallised.<\/p>\n\n\n\n<blockquote class=\"vf-blockquote\"><p>This is it, this is the one!<\/p><\/blockquote>\n\n\n\n<p>But the lines he saw were fuzzy. Dubochet lept from his chair, said something in French (which McDowall didn\u2019t understand) and ran through the lab. Although Dubochet\u2019s next declaration was in English\u2014\u201cThis is it, this is the one!\u201d\u2014McDowall already understood that they had finally succeeded in outpacing the ice crystals. The mysterious substance was the vitrified water they\u2019d spent two years chasing.<\/p>\n\n\n\n<p>Since then, the field of cryo-electron microscopy has, in McDowall\u2019s words, \u201cgone ballistic\u201d. Further developments have made it possible to image single particles, which has allowed researchers to better understand the structure of viruses such as Zika and Ebola, as well as the Tau protein, implicated in Alzehimer\u2019s disease. For his contribution to sample preparation for cryo-electron microscopy, <a href=\"https:\/\/news.embl.de\/science\/jacques-dubochet-awarded-nobel-prize-for-chemistry\/\" rel=\"noopener\">Dubochet has been awarded the Nobel Prize in Chemistry<\/a> while McDowall, now equipped with a doctorate, is in the process of setting up his third cryo-electron microscopy lab\u2014this latest one at Caltech.<\/p>\n\n\n\n<p>But despite all the changes to cryoelectron microscopy over the years\u2014improvements in cameras and computers and the increasing use of robots to image samples\u2014McDowall notes one constant.<\/p>\n\n\n\n<p>\u201cEverybody has to vitrify the sample,\u201d he says, referring to the method he contributed to developing back in his time a research technician. \u201cThat\u2019s still step one.\u201d<\/p>\n","protected":false},"excerpt":{"rendered":"<p>How a research technician with a master\u2019s degree contributed to Nobel Prize-winning work<\/p>\n","protected":false},"author":56,"featured_media":11871,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"footnotes":""},"categories":[5,17593],"tags":[50,54,548,136,43,241,35],"embl_taxonomy":[],"class_list":["post-11869","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-alumni","category-people-perspectives","tag-biochemistry","tag-chemical-biology","tag-dubochet","tag-electron-microscopy","tag-heidelberg","tag-lennart-philipson-award","tag-structural-biology"],"acf":{"article_intro":"<p>How a research technician with a master\u2019s degree contributed to Nobel Prize-winning work<\/p>\n","related_links":[{"link_description":"A curious case of serendipity","link_url":"https:\/\/news.embl.de\/alumni\/a-curious-case-of-serendipity\/"}],"article_sources":false,"vf_locked":false,"featured":false,"color":"#007B53"},"embl_taxonomy_terms":[],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v26.2 - https:\/\/yoast.com\/wordpress\/plugins\/seo\/ -->\n<title>Alasdair McDowall\u2019s slow road to flash freezing | EMBL<\/title>\n<meta name=\"description\" content=\"Alasdair McDowall, a research technician in Jacques Dubochet\u2019s lab (1978-1987), played a key role in developing technology to vitrify water.\" \/>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/www.embl.org\/news\/alumni\/alasdair-mcdowall-slow-flash-freezing\/\" \/>\n<meta property=\"og:locale\" content=\"en_US\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Alasdair McDowall\u2019s slow road to flash freezing | EMBL\" \/>\n<meta property=\"og:description\" content=\"Alasdair McDowall, a research technician in Jacques Dubochet\u2019s lab (1978-1987), played a key role in developing technology to vitrify water.\" \/>\n<meta property=\"og:url\" content=\"https:\/\/www.embl.org\/news\/alumni\/alasdair-mcdowall-slow-flash-freezing\/\" \/>\n<meta property=\"og:site_name\" content=\"EMBL\" \/>\n<meta property=\"article:publisher\" content=\"https:\/\/www.facebook.com\/embl.org\/\" \/>\n<meta property=\"article:published_time\" content=\"2017-12-08T10:52:48+00:00\" \/>\n<meta property=\"article:modified_time\" content=\"2024-03-22T10:51:52+00:00\" \/>\n<meta property=\"og:image\" content=\"https:\/\/www.embl.org\/news\/wp-content\/uploads\/2017\/12\/171208-mcdowall1.jpg\" \/>\n\t<meta property=\"og:image:width\" content=\"620\" \/>\n\t<meta property=\"og:image:height\" content=\"425\" \/>\n\t<meta property=\"og:image:type\" content=\"image\/jpeg\" \/>\n<meta name=\"author\" content=\"Sarah B. 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