Encapsulation of Algal Cells in Microdroplets for Cultivation and Screening
Lead Research Organisation:
University of Cambridge
Department Name: Chemistry
Abstract
The aim of this PhD project will be to develop a high-throughput microfluidic platform for screening algae cells. This will focus on two strains of algae, Phaeodactylum tricornutum (Pt) and Nannochloropsis gaditana (Ng). The first step will be to incubate algae cells in microdroplets and then track the growth of individual algal cells using fluorescence. This will inform on how algal cells behave in a microfluidic device. The cells will them be sorted by fluorescence activated droplet sorting to isolate key subpopulations.
Once this has been established attention will move to identification of transformant algae lines, using different fluorescent proteins (EGFP, EYFP, ECFP, etc.) as markers. By using this method, the transformant cells will be recovered directly from a cell population that has been electroporated, avoiding the process of waiting several weeks for visible colonies to appear on agar plates, and thus accelerating the algal engineering pipeline.
A further focus will be to develop a co-culture microdroplet method with the aim of culturing algae cells with other microorganisms. Some bacterial species have been shown to exchange nutrients with the algae, and the aim will be to examine this in microdroplets. The intensity of chlorophyll fluorescence or protein fluorescence can be used for an indirect way to monitor the growth of algae cells in microdroplets.
Once this has been established attention will move to identification of transformant algae lines, using different fluorescent proteins (EGFP, EYFP, ECFP, etc.) as markers. By using this method, the transformant cells will be recovered directly from a cell population that has been electroporated, avoiding the process of waiting several weeks for visible colonies to appear on agar plates, and thus accelerating the algal engineering pipeline.
A further focus will be to develop a co-culture microdroplet method with the aim of culturing algae cells with other microorganisms. Some bacterial species have been shown to exchange nutrients with the algae, and the aim will be to examine this in microdroplets. The intensity of chlorophyll fluorescence or protein fluorescence can be used for an indirect way to monitor the growth of algae cells in microdroplets.
Organisations
People |
ORCID iD |
| Chris Abell (Primary Supervisor) | |
| Tina Leontidou (Student) |
Studentship Projects
| Project Reference | Relationship | Related To | Start | End | Student Name |
|---|---|---|---|---|---|
| BB/M011194/1 | 01/10/2015 | 31/03/2024 | |||
| 1838346 | Studentship | BB/M011194/1 | 01/10/2016 | 31/12/2020 | Tina Leontidou |
| Description | A droplet microfluidic platform was established, which enables the screening of algal cells based on their chlorophyll content. The algal cells were encapsulated in microdroplets and then the droplet sorting platform was used to detect the chlorophyll fluorescence emitted by individual cells and to isolate droplets with higher fluorescence. The ability of the platform to distinguish between cells with high chlorophyll content and cells with low chlorophyll content was successfully tested by detecting the chlorophyll fluorescence of P. Tricornutum cells cultured under different light conditions. The results were presented at the EMBL Microfluidics 2018 conference through a poster presentation. Algal cells encapsulated in microdroplets were stained using BODIPY 505/515, a lipophilic dye. The staining enabled the detection of the lipid content of the cells. Quantitative detection of the lipid content of the cells with the microfluidic droplet sorting platform was not possible due to high background fluorescence levels. Currently, different methods are tested to reduce the background fluorescence and enable more accurate detection of the intracellular lipid content. |
| Exploitation Route | A platform has been established which enables high throughput screening of algal cells. Further down the line, this platform could be used by researchers, or potentially the industry, to identify algal cell strains that are most suitable for different biotechnological applications. |
| Sectors | Agriculture, Food and Drink,Energy,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology |