{"id":42694,"date":"2021-10-22T10:00:00","date_gmt":"2021-10-22T08:00:00","guid":{"rendered":"https:\/\/www.embl.org\/news\/?p=42694"},"modified":"2024-03-22T15:07:00","modified_gmt":"2024-03-22T14:07:00","slug":"analysis-and-sorting-with-flow-cytometry","status":"publish","type":"post","link":"https:\/\/www.embl.org\/news\/lab-matters\/analysis-and-sorting-with-flow-cytometry\/","title":{"rendered":"Analysis and sorting with flow cytometry"},"content":{"rendered":"\n

Throughout the planet\u2019s oceans reside a modest group of organisms with a fairly monumental task.<\/p>\n\n\n\n

These special bacteria, known as diazotrophs, convert atmospheric nitrogen into the biologically usable form that is essential for all organisms to live. As oceans warm and their pH balances change, scientists want to know more about these small organisms that make such a big contribution to our survival as a species. In many instances, the first step of these investigations is done with a flow cytometer.<\/p>\n\n\n\n

That is certainly the case for Hugo Berthelot, a postdoc in the Pepperkok Team who is studying the diverse shapes of diazotrophs. He is interested in whether their shape affects how well diazotrophs convert nitrogen or their capacity to withstand climate change in their habitat.<\/p>\n\n\n\n

Berthelot is also one of an increasing number of EMBL researchers whose work at the EMBL flow cytometry facilities goes beyond simply analysing and sorting human cells to reflect a broadening One Health perspective.<\/p>\n\n\n\n

\u201cWe\u2019re just at the beginning of understanding the diversity within these cells,\u201d Berthelot said. He uses the flow cytometer to sort cells before subjecting them to genomic and other omics analyses \u2013 to further understand, for example, where in the cells photosynthesis happens.<\/p>\n\n\n\n

The \u2018fluidity\u2019 of a flow cytometer <\/strong><\/strong><\/h2>\n\n\n\n

Cytometers and cell sorters use laser beams to analyse, sort, and separate cells or organisms flowing in a stable stream of fluid.<\/p>\n\n\n\n

\u201cImagine shining a beam of light through a stream of water as droplets pass by,\u201d said Diana Ordonez, the head of the Flow Cytometry Core Facility<\/a> at EMBL Heidelberg.<\/p>\n\n\n\n

\u201cThe instrument generates droplets containing particles or cells that are excited by the laser beams, measures the emitted light, and then deflects the drop carrying a particle of interest out of the stream and into a collection tube or plate,\u201d she explained. \u201cThat\u2019s what makes it possible to separate out a given particle.\u201d<\/p>\n\n\n\n

For instance, scientists could separate a population of interest, such as T cells, from a blood sample containing several types of immune cells and use it for downstream analysis.<\/p>\n\n\n\n

\"Male
\u201cMy hope is to be actively involved in projects here,\u201d said Gerald Pfister, who is the head of the Flow Cytometry facility at EMBL Rome. Credit line: Rossana De Lorenzi\/EMBL<\/figcaption><\/figure><\/div>\n\n\n\n

Ordonez and Gerald Pfister, the head of the Flow Cytometry Facility in EMBL Rome\u2019s Epigenetics and Neurobiology Unit<\/a>, agree that if a sample can be suspended in a liquid, a flow cytometer can analyse it.<\/p>\n\n\n\n

\u201cThe most classic example is when you have blood test; cell populations in your blood are identified and quantified using a flow cytometer,\u201d Pfister said. \u201cBut flow cytometers are used for looking at a variety of things \u2013 algae cells, ciliates (protozoans with hairy edges known as cilia), bacteria in water samples, even in food quality control systems to assess bacteria in milk or yeast cells in beer.\u201d<\/p>\n\n\n\n

Flow cytometry and building bridges<\/strong><\/strong><\/h2>\n\n\n\n

Ordonez, who has worked at EMBL Heidelberg\u2019s flow cytometry facility since 2015, took the helm in February.<\/p>\n\n\n\n

Pfister came to EMBL Rome\u2019s facility in June after more than 20 years in different roles in industry and academia around the world, from Austria and Brazil to Belgium to Qatar. In Qatar, he spearheaded the development of a flow cytometer core lab. In all his roles, he has been mindful of end-user service as well as public outreach.<\/p>\n\n\n\n

\u201cMy hope is to be actively involved in projects here,\u201d Pfister said. \u201cAt this point, I have a lot of tips and ideas, so I can really contribute as people plan projects.\u201d<\/p>\n\n\n\n

Ordonez also has a long history with flow cytometry. Before coming to EMBL, she used flow cytometry at the Centre d’immunologie de Marseille-Luminy to study a variety of immune cells \u2013 T cells, B cells, dendritic cells, natural killer cells, monocytes, and progenitor cells \u2013 in mice. Her goal was to establish a reliable model to analyse the role that a specific kind of white blood cell has in the immune response.<\/p>\n\n\n\n

\"Female
Diana Ordonez has worked at EMBL Heidelberg\u2019s flow cytometry facility since 2015 but began her leadership role there in February. Credit: Stuart Ingham\/EMBL<\/figcaption><\/figure><\/div>\n\n\n\n

She then moved to EMBL, principally because \u2013 like Pfister \u2013 she enjoys training, advising, and assisting other researchers in flow cytometry. It\u2019s this attitude that is a guiding force for both scientists as they share their plans for the future.<\/p>\n\n\n\n

Looking ahead<\/strong><\/strong><\/p>\n\n\n\n

As EMBL embarks on its next five-year research programme, EMBL\u2019s flow cytometry facilities are looking at ways to adapt to an increasingly multidisciplinary environment, and one that will begin exploring living systems in their natural context.<\/p>\n\n\n\n

\u201cI have a PhD in freshwater biology, so I am looking forward to this challenge,\u201d Pfister said. \u201cThis opens up opportunities to collaborate with different research teams, not just on biomedicine or related to medical conditions.\u201d<\/p>\n\n\n\n

The two EMBL facilities have flow cytometers and cell sorters with slightly different capabilities. Ordonez and Pfister perform most of the flow cytometry work themselves, advising individual researchers on their given projects, developing optimal flow applications, and troubleshooting their experimental setup.<\/p>\n\n\n\n

Training helps to expand the pool of hands-on users and allows others to learn how flow cytometry can be used in their own varied research projects. From data analysis workshops to practical courses in flow cytometry and cell sorting presented in cooperation with the German Cancer Research Center (DFKZ), EMBL\u2019s flow cytometry training is offered to the international science community and is always in high demand. It\u2019s why Ordonez is looking for creative ways to expand it.<\/p>\n\n\n\n

Teamwork makes the flow cytometry work\u2026better<\/strong><\/strong><\/h2>\n\n\n\n

Collaborations with other EMBL core facilities have been part of that creative management approach in the past, so will likely play a role in the future. Ordonez is quick to note the convenient location of the Genomics Core Facility (GeneCore), which is right across the hall doing gene sequencing \u2013 a logical follow-up to flow cytometry for epigenetics and genetics researchers.<\/p>\n\n\n\n

Over the years, the challenging scientific needs of EMBL\u2019s flow cytometry customers have made these facilities a good testbed for piloting new technology. In fact, Berthelot\u2019s research benefits from exactly this kind of \u2018beta testing\u2019 of a new instrument. He is using detectors to create rapid-fire images and use them in addition to conventional flow cytometry parameters to analyse and separate particles of interest. The images help Berthelot to more easily identify and understand the complexity of organisms present in his samples.<\/p>\n\n\n\n

Ordonez and Pfister agree that the two facilities will increasingly find ways to collaborate to enhance their offerings and position themselves better at EMBL. \u201cI started out studying microbiology because I had this dream to do oil-spill remediation,\u201d Ordonez recounted. \u201cThough I ended up studying immunology, we all need to do something for this planet. It just has to be done. All these resources and bright minds here at EMBL can make a difference.\u201d<\/p>\n","protected":false},"excerpt":{"rendered":"

A technology around since the \u201860s, flow cytometry has increasing applications. New leadership at EMBL\u2019s flow cytometry facilities is looking to ease use, expand training, and encourage more collaboration.<\/p>\n","protected":false},"author":100,"featured_media":43458,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"footnotes":""},"categories":[3,17591],"tags":[580,64,227,1704,43,33,382,514,76],"embl_taxonomy":[9788,3742,10128,19089],"class_list":["post-42694","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-lab-matters","category-science-technology","tag-bacteria","tag-cell-biology","tag-core-facility","tag-embl-programme","tag-heidelberg","tag-instrumentation","tag-plankton","tag-rome","tag-service","embl_taxonomy-embl-rome","embl_taxonomy-flow-cytometry-core-facility","embl_taxonomy-flow-cytometry-facility","embl_taxonomy-rainer-pepperkok"],"acf":{"featured":true,"show_featured_image":false,"field_target_display":"","article_intro":"

EMBL\u2019s flow cytometry facilities support a variety of life sciences research, with new leaders planning to enhance their offerings<\/p>\n","related_links":[{"link_description":"eLife: Core strengths: an interview with Diana Ordonez","link_url":"https:\/\/elifesciences.org\/interviews\/bc173bec\/diana-ordonez"},{"link_description":"EMBL Flow Cytometry Core Facility","link_url":"https:\/\/www.embl.org\/groups\/flow-cytometry-heidelberg\/"},{"link_description":"Flow Cytometry Facility in EMBL Rome\u2019s Epigenetics and Neurobiology unit","link_url":"https:\/\/www.embl.org\/groups\/flow-cytometry-rome\/"},{"link_description":"Go with the flow","link_url":"https:\/\/www.embl.org\/news\/science\/1410_flow\/"}],"source_article":false,"in_this_article":false,"press_contact":"None","vf_locked":false},"embl_taxonomy_terms":[{"uuid":"a:3:{i:0;s:36:\"b14d3f13-5670-44fb-8970-e54dfd9c921a\";i:1;s:36:\"89e00fee-87f4-482e-a801-4c3548bb6a58\";i:2;s:36:\"741d5d3d-f92f-4eb8-9195-55c96454a36b\";}","parents":[],"name":["EMBL Rome"],"slug":"embl-rome","description":"Where > All EMBL sites > EMBL Rome"},{"uuid":"a:3:{i:0;s:36:\"302cfdf7-365b-462a-be65-82c7b783ebf7\";i:1;s:36:\"ef0437fc-a5b7-4c73-bcfd-63bff16cb35e\";i:2;s:36:\"53113f1a-14eb-4788-b0e1-0f89a00845cb\";}","parents":[],"name":["Flow Cytometry Core Facility"],"slug":"flow-cytometry-core-facility","description":"What > Services and facilities > Flow Cytometry Core Facility"},{"uuid":"a:3:{i:0;s:36:\"302cfdf7-365b-462a-be65-82c7b783ebf7\";i:1;s:36:\"ef0437fc-a5b7-4c73-bcfd-63bff16cb35e\";i:2;s:36:\"7fc947bf-6dee-44d4-8280-e30a14ab129e\";}","parents":[],"name":["Flow Cytometry Facility"],"slug":"flow-cytometry-facility","description":"What > Services and facilities > Flow Cytometry Facility"},{"uuid":"a:2:{i:0;s:36:\"4428d1fd-441a-4d6d-a1c5-5dcf5665f213\";i:1;s:36:\"b75fccdd-f939-4989-9803-f876975537d3\";}","parents":[],"name":["Rainer Pepperkok"],"slug":"rainer-pepperkok","description":"Who > Rainer Pepperkok"}],"yoast_head":"\nAnalysis and sorting with flow cytometry | EMBL<\/title>\n<meta name=\"description\" content=\"A technology around since the \u201860s, flow cytometry has increasing applications. 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