Song, Jiyoung; Ryu, Hyunryul; Chung, Minhwan; Kim, Youngtaek; Blum, Yannick; Lee, Sung Sik; Pertz, Olivier; Jeon, Noo Li (2018). Microfluidic platform for single cell analysis under dynamic spatial and temporal stimulation. Biosensors & bioelectronics, 104, pp. 58-64. Elsevier 10.1016/j.bios.2017.12.038
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Recent research on cellular responses is shifting from static observations recorded under static stimuli to real-time monitoring in a dynamic environment. Since cells sense and interact with their surrounding microenvironment, an experimental platform where dynamically changing cellular microenvironments should be recreated in vitro. There has been a lack of microfluidic devices to support spatial and temporal stimulations in a simple and robust manner. Here, we describe a microfluidic device that generates dynamic chemical gradients and pulses in both space and time using a single device. This microfluidic device provides at least 12h of continuous stimulations that can be used to observe responses from mammalian cells. Combination of the microfluidic de-vice with live-cell imaging facilitates real-time observation of dynamic cellular response at single cell level. Using stable HEK cells with biosensors, ERK (Extracellular signal-Regulated Kinase) activities were observed un-der the pulsatile and ramping stimulations of EGF (Epidermal Growth Factor). We quantified ERK activation even at extremely low EGF concentration (0.0625µg/ml), which can not be observed using conventional techniques such as western blot. Cytoskeleton re-arrangement of the 3T3 fibroblast (stable transfection with Lifeact-GFP) was compared under abrupt and gradually changing gradient of PDGF.
Item Type: |
Journal Article (Original Article) |
|---|---|
Division/Institute: |
08 Faculty of Science > Department of Biology > Institute of Cell Biology |
UniBE Contributor: |
Pertz, Olivier |
Subjects: |
500 Science > 570 Life sciences; biology |
ISSN: |
0956-5663 |
Publisher: |
Elsevier |
Language: |
English |
Submitter: |
Olivier Pertz |
Date Deposited: |
19 Feb 2018 14:04 |
Last Modified: |
05 Dec 2022 15:09 |
Publisher DOI: |
10.1016/j.bios.2017.12.038 |
PubMed ID: |
29306762 |
Uncontrolled Keywords: |
Chemotaxis Dynamic gradient ERK kinetics Long-term live cell imaging Microfluidic Single cell analysis Temporal stimulation |
BORIS DOI: |
10.7892/boris.109210 |
URI: |
https://boris.unibe.ch/id/eprint/109210 |
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