Microfluid incubator

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Labratory Notebook

Click here for Lab and Planning Notes


Spring Semester 2022

This semester we have worked hard in order to achieve control over CO2 concentration inside the incubator in order to prolong the life of the cells. We have received Zimmer & Peacock ISE OEM, pH sensor and software which we needed to integrate into our experiment in order to measure the pH concentration.

We have used some time to create new wafers with a new design, which we successfully used in our experiments.

PH sensor

When trying to integrate the Zimmer & Peacock pH sensor we did not receive the results we were hoping for. The issue of integrating the sensor together with the medium flow is still under planning stage.

Timeline for Spring 2022:

27.01.2022 - Kick off meeting to start off the new semester. We have discussed our goals and how to divide the project between the participants for more efficiency. Discussing how to solve the issues with bubbles and how to control the CO2 levels. The issues which have not yet been resolved in one place.

08.03.2022 - First Photolithography session since Spring 2021. After the procedure we have succesfully created new wafers which we used in upcoming experiments. All you want to know about Photolithography.

04.01.2022 - First cell adhesion experiment has been divided in 4 parts: Bonding, filling the wafer with water, filling it with fibronectin and in the end introducing the cells. Keep in mind that as the experiment proceeded, we created a new protocol for these types of procedures by trial and error which can be time consuming. The experiment did not complete stage 4 due to a mistake in stage 1.

04.04.2022 - Second cell adhesion experiment. Similar as the first cell adhesion experiment. This time stage 4 was completed and cells were filled into the chambers, but no follow up data is available due to an error with the miscroscope.

Autumn Semester 2021

This semester there has been only two trials of cell passaging. The first trial included the preparation of the microfluidic chip together with fibronectin aliquoting and injection of the cells. Some of the injected cells had successfully adhered to the fibronectin coated surface. The second trial was similar, with a few of the cells attaching to the surface. The end of this semester was dedicated to creating a new team of students which would continue to work together in the following semester.

Progression Summer 2021

The participants showed huge interest in the project and decided to continue working on it throughout the summer! This time around the team has used collagen to coat the surface of the chip instead of fibronectin.

The number one enemy for them were the air bubbles, which unfrotunately were killing the cells while entering through the stream and drying them out. In order to solve this issue, they have created a plan to restrict bubbles from coming in and wrote about in their entry called Microfluidics Bubble Genocide! In short, they used a syringe already filled with fluid and connected it to the outlet in order to create a backflow, so that the fluid will get sucked inside. They also made sure not to use high pressure, since it can create leaks which can cause bubbles. After doing this they had no sign of bubbles. Durng this time there has been 9 cell adhesion trials.

Progression Spring 2021

This semester consisted of a lot of meetings and planning in order to work out correct procedures for the experiments. There were 2 cell adhesion trials this semester.

Microfluid monitoring incubator

In depth information about the project

Here is a presentation of the concept

Check out this introduction to Cell biology: Microfluidic Concepts and methodologies

Check out Microfluidics for Cell Biology

Scientific paper on following topic [add papers here]

The rage of the 2020s are organs-on-a-chip [1]. These are systems that have to be specifically engineered for each type of organ. Growing and maintaining cell cultures, however is a more generic activity of cell biology. They are kept in expensive incubators and several times a week they have to be looked after and the growth media replaced. When there are too many cells, most of them are killed and some are re-applied in a new flask/petri dish. All this can be done inside a lab on a chip[4]. In addition, the incubator functions of controlling temperature, CO2 and O2 pressure can be reduced to small modules in a standalone cell culture system that is small enough to be moved from lab to lab and between research microscopes at different imaging facilities. Adding electrodes to the system will allow electrochemical or bioimpedance monitoring of the cell coverage, a feature that will be publishable in a scientific journal.

The system is perhaps not the most exciting project to start with, but it contains all the basic technology needed for more advanced organ-on-a-chip systems: temperature, gas and fluid control, hydrogel membranes, monitoring of cell state, etc. It will be a platform that LagLivLab students can build on and modify for years.

[1] B. Zhang, A. Korolj, B.F.L. Lai, M. Radisic, Advances in organ-on-a-chip engineering, Nat. Rev. Mater. 3 (2018) 257–278. doi:10.1038/s41578-018-0034-7.
[4]	F. Bunge, S. van den Driesche, M.J. Vellekoop, Microfluidic platform for the long-term on-chip cultivation of mammalian cells for Lab-on-a-Chip applications, Sensors (Switzerland). 17 (2017) 1–15. doi:10.3390/s17071603.