University of Twente Student Theses
The influence of fluid flow on trapped protoplast in a microfluidic device
Zijlstra, M.A. (2024) The influence of fluid flow on trapped protoplast in a microfluidic device.
|
PDF
37MB |
| Abstract: | Protoplasts are plant cells with their cell wall enzymatically removed, and therefore quite fragile. In this report, a protocol developed for trapping protoplasts (isolated of the leaves of Nicotiana Tabacum) inside a microfluidic device will be discussed. A chip is made of polydimethylsiloxane (PDMS) with U-shaped trapping structures, bonded to a microscope glass slide. No air bubbles on chip were established after filling the chip with fluid directly after bonding (using an oxygen plasma oven). When varying the flow speed, the most efficient trapping was reached with a flow speed of 2.0 μL/min. A protocol was made for the research into the influence of continuous flow on trapped protoplasts. Due to unforeseen external interference, these experiments failed. Thence, the influence of penicillin-streptomycin (pen/strep) on the viability of protoplasts was tested. Based on the morphology of the protoplasts, It seems that pen/strep does not harm the protoplast. However, more research is needed into the influence on cell wall regrowth and viability over a longer time. The protocol for continuous flow on trapped protoplasts seems promising based on morphology during the monitoring of the experiments. To quantify the viability of the protoplasts, the use of live/dead staining Fluorescein Diacetate (FDA) and propidium iodide (PI) is evaluated. On day 0, FDA staining evaluates the viability of the protoplasts, but harms the protoplasts, resulting in dead protoplasts after 24 hours. The results of this project can be used to trap protoplasts on chip and evaluate the viability for several days, but further research is needed to optimize the used protocols. |
| Item Type: | Essay (Bachelor) |
| Faculty: | TNW: Science and Technology |
| Subject: | 30 exact sciences in general, 42 biology, 48 agricultural science |
| Programme: | Biomedical Technology BSc (56226) |
| Link to this item: | https://purl.utwente.nl/essays/99894 |
| Export this item as: | BibTeX EndNote HTML Citation Reference Manager |
Show download statistics for this publication
Show download statistics for this publicationRepository Staff Only: item control page