A Single-Cell Study of a Highly Effective Hog1 Inhibitor for in Situ Yeast Cell Manipulation
Artikel i vetenskaplig tidskrift, 2014

We present a single cell study of a highly effective Hog1 inhibitor. For this application, we used sequential treatment of a Saccharomyces cerevisiae cell array, with the Hog1 inhibitor and osmotic stress. For this purpose, a four-inlet microfluidic chamber with controlled introduction of two different cell strains within the same experimental setting and a subsequent rapid switching between treatments was designed. Multiple cell strains within the same experiment is a unique feature which is necessary for determining the expected absent cellular response. The nuclear translocation of the cytosolic MAPK, Hog1, was monitored by fluorescence imaging of Hog1-GFP on a single-cell level. An optical tweezers setup was used for controlled cell capture and array formation. Nuclear Hog1-GFP localization was impaired for treated cells, providing evidence of a congenial microfluidic setup, where the control cells within the experiments validated its appropriateness. The chamber enables multiple treatments with incubation times in the order of seconds and the possibility to remove either of the treatments during measurement. This flexibility and the possibility to use internal control cells ensures it a valuable scientific tool for unraveling the HOG pathway, similar signal transduction pathways and other biological mechanisms where temporal resolution and real time imaging is a prerequisite.

optical trapping

cell-to-cell variability

Saccharomyces cerevisiae

HOG

optical tweezers

MAPK

single-cell analysis

system biology

inhibitor

microfluidics

Författare

Charlotte Hamngren Blomqvist

SuMo Biomaterials

Göteborgs universitet

Chalmers, Teknisk fysik, Eva Olsson Group

Peter Dinér

Göteborgs universitet

Morten Grötli

Göteborgs universitet

Mattias Goksör

Göteborgs universitet

Caroline B. Adiels

Göteborgs universitet

Micromachines

2072-666X (ISSN)

Vol. 5 1 81-96 mi5010081

Ämneskategorier

Nanoteknik

DOI

10.3390/mi5010081