Corporate Banner
Satellite Banner
Spectroscopy
Scientific Community
 
Become a Member | Sign in
Home>News>This Article
  News
Return

Investigating Protein Targets and Cellular Pathways in Yeasts

Published: Monday, October 07, 2013
Last Updated: Monday, October 07, 2013
Bookmark and Share
Scientists at the Donnelly Centre for Cellular and Biomolecular Research have chosen generations of Tecan microplate readers to monitor the effects of environmental or drug perturbation on molecularly barcoded yeasts.

The Donnelly Centre for Cellular and Biomolecular Research, based at the University of Toronto, Canada, is an interdisciplinary research institute that houses scientists from a wide variety of backgrounds, integrating the fields of biology, computer science, engineering and chemistry, as well as leading areas of biomedical research. Corey Nislow, Associate Professor at the Donnelly Centre’s Banting and Best Department of Medical Research (BBDMR) faculty, explained: “The Donnelly Centre houses faculty from many different departments, adopting a multidisciplinary approach to experimental studies with the intention of cross-pollinating between bioinformaticians, genomic and proteomic scientists. My own laboratory, and the laboratory of my collaborator Dr Guri Giaever, is responsible for running a next generation chemogenomics facility, and our work involves monitoring the growth of large pools of molecularly barcoded yeast, simultaneously screening 6,000 different mutants. The presence of a unique mutant barcode identifier enables us to distinguish each one of the mutants and deconvolute a complex sample pool at the end of an experiment. This simplifies the procedure, allowing the pool to be treated as a simple culture and challenged with different environmental or drug perturbations.” 

Corey continued: “Guri, Michael Proctor (a research scientist working at Stanford University at the time) and myself  first developed a workstation for this work in 2000, consisting of four Tecan GENios™ microplate readers to monitor the growth of the culture, integrated with a Perkin Elmer multiprobe. The basic principle is the same for this and the second generation system we later created using Tecan’s Safire™ reader and Freedom EVO® 200 liquid handling workstation. Based on a predetermined parameter, the liquid handler samples and reinoculates the culture, keeping it in logarithmic phase for up to 100 generations and allowing subtle effects on different mutants to be investigated. Initially, the liquid handler moves some of the culture to fresh media and, at the time of transfer, saves a sample so that the abundance of each strain can be decoded at the end of the experiment by microarray hybridization or next generation sequencing. The readers monitor both optical density and fluorescence – 95 % of the time we monitor optical density, but occasionally we study the readout from a fluorescent reporter – and each well of each plate is independently monitored. Essentially, we chart the abundance of every strain under a particular condition and, based on the abundance of the different mutants, infer the particular protein targets or cellular pathways that are important for culture survival under those circumstances. In the absence of these protein targets or cellular pathways, the culture is sensitive to that particular condition.” 

“For the first system we chose the GENios reader because it was the only instrument at the time that had a sufficient orbit to keep yeast cells well suspended and that could maintain temperature without condensation. However, although this system is still in use, the workflow means that the readers are effectively serving as shaking incubators, operating 24 hours a day, and shaking and reading at 15 minute interviews. The process of ejecting each plate to allow the liquid handler to sample the culture and return the plate takes a minute each time and is very much a rate-limiting step. In contrast, the second generation screening system is equipped with six shaking incubators, and the Safire microplate reader is just used to read! The speed of the Safire reader allows six to eight plates to be accommodated without any additional waiting time, and we can now interrogate model organisms that require light, such as the model algae Chlamydomonas; this would previously have been impossible in the dark of the reader. In addition, using dedicated shaking incubators has enabled us to increase capacity.”

“We didn’t just ‘settle on’ Tecan, we purposely selected the Company, and so did all of our collaborators, who loved the growth curves they saw from our work. Every time we ran large screens we needed to confirm individual strains, and that required a server rack full of readers. We have remained with Tecan and, between our Stanford site and Toronto, have 24 GENios systems, which are all still running! The flexibility of the Tecan systems is a big advantage, particularly the smooth information transfer.”

“As well as increasing throughput, our Tecan instruments have given us new avenues of exploration. We are now focusing on data collection and the introduction of new organisms, developing simple barcoding strategies for other organisms and investigating E. coli strains associated with Crohn’s Disease. We also plan to study more model organism genomes, de novo genomes, and will be doing a lot of next generation sequencing library preparation. In the future, we plan a third generation system, with two new generation Tecan readers and twelve shaking incubators, which will further increase our capabilities,” concluded Corey.


Further Information
Access to this exclusive content is for Technology Networks Premium members only.

Join Technology Networks Premium for free access to:

  • Exclusive articles
  • Presentations from international conferences
  • Over 2,500+ scientific posters on ePosters
  • More than 3,700+ scientific videos on LabTube
  • 35 community eNewsletters


Sign In



Forgotten your details? Click Here
If you are not a member you can join here

*Please note: By logging into TechnologyNetworks.com you agree to accept the use of cookies. To find out more about the cookies we use and how to delete them, see our privacy policy.

Related Content

Too Gracious a Host
Scientists at the University of Toronto are using Tecan’s Infinite M200 PRO and Gas Control Module to study host-pathogen interactions for Legionella bacteria.
Monday, October 07, 2013
A Watchful Eye on Biosecurity
Intended for government laboratory and pharmaceutical screening applications, BioSentinel now recommends the Infinite F500 to all its customers.
Monday, October 07, 2013
Scientific News
A Highly Sensitive Graphene Based Sensor
Researchers at EPFL and ICFO have developed a sensor made from graphene to detect molecules such as proteins and drugs.
Cannabis May Be Used to Treat Fractures
TAU researcher finds non-psychotropic compound in marijuana can help heal bone fissures.
Researchers Reveal Elusive Molecule
A long-standing chemistry puzzle has been solved, with potential implications ranging from industrial processes to atmospheric chemistry.
Optical 'Dog's Nose' Developed to Detect Cancer, Other Diseases
Researchers are using optical spectroscopy to develop a quick, non-invasive “breath test” they believe will have the potential to screen for a variety of diseases, including diabetes, infections and cancers.
Unravelling the Mysteries of Carbonic Acid
Researchers have shown how gaseous carbon dioxide molecules are solvated by water to initiate the proton transfer chemistry that produces carbonic acid and bicarbonate.
NIR Spectroscopy Produces a Handy Image of Blood Circulation
Poor blood circulation can be revealed by a novel form of near infrared (NIR) spectroscopy, say Italian medical researchers.
Combination Imaging Reveals Fuel Cell Damage
A simultaneous view of both chemical distribution and bonding states in fuel cell membranes shows how and where irreversible degradation takes place.
Shining A New Light On The Immune System
Scientists at the University of St Andrews have developed a revolutionary method of identifying cells of the immune system with “molecular fingerprints” which could pave the way for the rapid detection of conditions such as leukaemia and lymphoma from a small blood sample.
Holes in Gold Enhance Molecular Sensing
Electrochemical techniques produce tuneable porous gold films, where the empty spaces enhance light scattering and sensing signals.
Damming hemorrhagic diseases
A potential mechanism to combat diseases caused by haemorrhagic fever viruses has been discovered by researchers.
Skyscraper Banner

Skyscraper Banner
Go to LabTube
Go to eposters
 
Access to the latest scientific news
Exclusive articles
Upload and share your posters on ePosters
Latest presentations and webinars
View a library of 1,800+ scientific and medical posters
2,500+ scientific and medical posters
A library of 2,500+ scientific videos on LabTube
3,700+ scientific videos
Close
Premium CrownJOIN TECHNOLOGY NETWORKS PREMIUM FREE!