Utskrift från Malmö högskolas webbplats www.mah.se

Development of new cell-based in vitro techniques using digital holography and phase contrast microscopy

Contact person: Anette Gjörloff Wingren
Responsible: Anette Gjörloff Wingren
Co-workers: Tautgirdas Ruzgas
Partner: Phase Holographic Imaging AB, Lund, Christer Wingren, Lund
Funding: Magnus Bergwalls stiftelse, UMAS cancerstiftelse
Timeframe: 2010-01-01 -- 2014-12-31
Research profile: Biologiska gränsytor
Research programme: Biofilms Research Center for Biointerfaces
Faculty/Department: Faculty of Health and Society, The Department of Biomedical Science
Subject: Medicin
Webpage: http://www.mah.se/anettegw

We are using the novel label-free technique of digital holography to study cellular parameters such as cell number, confluence, cell area and thickness in cultured cells. We have investigated whether cell death induced cells can be distinguished from untreated control cells by using this technique. The analysis was performed directly in the cell culture flasks without any de-attachment of the cells. Adherent cancer cell lines were treated with anti-tumor agents, or left untreated, for up to 3 days. Measurements by digital holography showed differences in cell number, confluence, cellular area and thickness. The analysis was followed by a flow cytometric analysis of the same cells, determining AnnexinV-FITC labelling together with the proportion of propidium iodide labelled cells. The results obtained with digital holography and flow cytometric analysis of viable and dead cells were highly comparable. Also, recombinant focused antibody microarrays (<10 antibodies) are fabricated on a glass substrate and can directly be interfaced with digital holography for label-free sensing of captured cells. Certain populations of leukemia and lymphoma cells will be sorted out by attracting the cells to the recombinant antibody-functionalized nano-areas. By combining cellular analysis with digital holography and antibody-targeted “cell sorting” we can determine how defined subpopulations of cells behave when treated by a specific compound. Another aim is to develop in-vitro methods for studies of cell surface interactions and artificial wound-healing assays. We want to understand the first steps in integration of implanted devices with tissues and oppositely the effect of long-term cellular systems on the life-time and performance of implanted devices, e.g., biosensor-biofuel cell for continuous monitoring purposes. We typically work with artificial wound-healing assays, which generally involve first the growing of a confluent cell monolayer, then electrical wounding of cell monolayer and monitoring its recovery. Future applications can include real-time cell monitoring of various parameters of cells in response to clinically relevant compounds. 

Beskrivning på svenska

Vi studerar  viabilitet och celldöd hos cancer celler med digital holografi, en helt ny teknik för att studera celler. Vi använder denna innovativa bildbehandlingsmetod för cellstudier, där cellerna ej förstörs under behandlingen, ej heller behöver märkas in med radioaktivitet eller antikroppar/färger. Vi studerar cellöverlevnad och celldöd (apoptos) hos cancer celler. Preliminära resultat visar att tekniken är jämförbar med konventionella apoptosmätningar som görs med hjälp av flödescytometri och mikroskop idag. Tekniken kommer också att för att jämföra olika typer av lymfom cancer celler och studera om digitala holografimätningar av cellstorlek, täthet, yta, tjocklek, etc är ett diagnostiskt verktyg. Ett annat mål är att utveckla in-vitro metoder för att studera cellulära interaktioner på olika ytor och artificiella ” wound-healing assays”.

Senast uppdaterad av Magnus Jando