CellASIC Products - ONIX for Yeast Cells

Microfluidic Flow Chamber for Yeast

The Y-Series microfluidic plates keep yeast cells in a single focal plane, allowing you to follow and induce cell events during high magnification imaging over many generations.

Features and Benefits:

• Elastic trap sites holds cells in single focal plane for 16+ hours
• 4 independent chambers under a single imaging window
• Laminar flow switching between 6 inlet solutions
• 170 um thick glass bottom for high quality imaging

Plate Types:

Y04C for Haploid Cells, 4 Chambers (3.5-5 um)
Y04D for Diploid Cells, 4 Chambers (5-7 um)

Downloads:

Informational Flyer
Yeast Genetics 2010 Poster
Y04 Application Note

Description:

The Y04 plates utilize a microfabricated silicone ceiling with a height similar to yeast cells to restrict their growth in a single focal plane and maintaining x,y position over time. Each chamber contains 3 tapered trap heights to maximize loading efficiency of various cell sizes. The Y04C plate has heights of 4.0, 4.5, and 5.0 um, and the Y04D plate has heights of 5, 6, 7 um. (See Product Sheet for more detail.)

Yeast Trap Yeast Focus

Continuous perfusion of culture medium to the cells ensures a suitable growth environment for long term experiments. Typical experiments run for 4-16 hours on the microscope stage, enabling single cell tracking for over 10 doublings. The six inlets to each chamber allow solution switching experiments during time lapsed imaging. The highly laminar flow results in a rapid and complete solution change in under a minute.

Yeast Growth

Applications:

• Cell Response to Media Change
• Induction of Cell Cycle Events
• GFP Linked Nuclear Trafficking

• Cell Division Tracking
• Starvation and Recovery
• Gene Expression

• Protein Localization
• Mitochondria Inheritance
• Phase Contrast, DIC

Movies:

1. Budding Yeast Growth - Cell division of S. cerevisiae with continuous flow.
2. Fission Yeast Growth - Cell division of S. pombe with continuous perfusion.
3. Fission Yeast Growth - Cell division of S. japonicus with continuous perfusion.
4. Signal Tracking - Monitoring of gene expression activity over generations.
5. Nuclear Shuttling - Nuclear transport in response to a switching extracellular signal.
6. Cell Loading - S. cerevisiae loading into the microfluidic device.
7. Solution Switching - Laminar flow exchange in the Y04 chamber.
8. Imaging with DIC Objective - S. cerevisiae growth imaged with 150X DIC objective.
9. Replication Fork - S. pombe replication fork stalling and recovery.
10. Cell Cycle Arrest and Release - S. cerevisiae exposed to alpha factor.
11. Aspergillus Growth - Perfusion culture of Aspergillus in the Y04C chamber.
12. Yeast Induction - GAL induction and repression in budding yeast

Image Gallery:

Click Here to View


S. cerevisiae, Lim Lab, UCSF	S. cerevisiae, Maheshri Lab, MIT


Mitochondiral structure, Marshall Lab, UCSF	Fission yeast, Niki Lab, NIG, Japan


Cell cycle tracking, Skotheim Lab, Stanford	Fission yeast, Forsburg Lab, USC


S. pombe, cells courtesy Forsburg Lab, USC	150X objective with DIC, Jan Wisniewski, NIH


Cell arrest and release, courtesy of Lacefield Lab, Univ. of Indiana

Testimonials:

Click Here to View

Research Lab:	Marshall Lab, UCSF (http://biochemistry.ucsf.edu/labs/marshall/)

Project Title:	Imaging yeast cells to quantify mitochondrial morphology

Authors:	SM Rafelski, WF Marshall

Testimonial:	"I use the Onix system with the Yeast plates routinely in my imaging of live, growing yeast cells to image the mitochondria in these cells. Since I aim to quantify mitochondrial morphology, I require constant, stable imaging conditions that maintain the health of the cells, which the Onix system does very well. In addition, the system allows me to perform pharmacological or nutrient-switching timelapse experiments. I have found that the growth properties of the same cells on different days is identical in these flow chambers, which is very helpful for collecting larger data sets. Cells can continue to grow and divide until they run out of space, and survive with renewing nutrients for 1-2 days. I have been using Cellasic plates since January, 2008 and the company has continually improved their plate design, vacuum manifold design, and added many new types of plates. They have also been exceedingly helpful in getting the system up and running and any troubleshooting that comes along."
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Research Lab:	Maheshri Lab, MIT (http://web.mit.edu/~narendra/www/)

Project Title:	Modification of Transcriptional Regulatory Networks in Budding Yeast

Authors:	CJ Zopf, N Maheshri

Testimonial:	“Our experience with the system and CellAsic has been excellent. We've been able to quickly and easily perform novel and technologically demanding experiments without any prior microfluidic experience. I've been able to focus on the fundamental biological questions while letting CellAsic provide me with the tools I need to answer them.” “CellASIC is continually improving the system. We have been using their devices through many generations and have seen vast improvement and consideration of the feedback we have provided.” “Large majority of cells loaded at the beginning of the experiment stay fixed in place for excellent time trace of single cells over long experiments (up to 16 hours in our case) Close trapping chamber positioning in Y4 reduce travel time for the stage, facilitating more frequent data acquisition. The system uses flow rates that can quickly switch media in the trapping chamber but are low enough that the reservoirs can provide for long experiments."

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Research Lab:	Skotheim Lab, Stanford (http://web.me.com/skotheim/Site/Welcome.html)

Project Title:	Cell Cycle Commitment

Authors:	A Doncic, JM Skotheim

Testimonial:	“If you are interested in a small/medium scale system for microfluidic single-cell analysis this is a very good system which we are extremely satisfied with. Easy to work with: both to prepare the plate and to work with the software which allows us to focus on experiments and not on ‘whether the flowcell will crash after X hours.’” “Excellent customer interaction/customer friendly representatives: before buying we were allowed to use the unit over a month to confirm that it is as good as they claimed. All-in-all it is an excellent product that we highly recommend.”

Publications:

Click Here to View

Bermejo C, Haerizadeh F, Takanaga H, Chermak D, Frommer W. "Optical sensors for measuring dynamic changes of cytosolic metabolite levels in yeast." Nature Protocols. 2011 October 27 6;1806-1817.

Doncic A, Falleur-Fettig M, Skotheim J. "Distinct interactions select and maintain a specific cell fate." Molecular Cell. 2011 Aug 19 4;43:528-539.

Eser U, Falleur-Fettig M, Johnson A, Skotheim J. "Commitment to a cellular transition precedes genome-wide transcriptional change." Molecular Cell. 2011 Aug 19 4;43:515-527.

Tamura N, Oku M, Sakai Y. "Atg8 regulates vacuolar membrane dynamics in a lipidation-independent manner in Pichia pastoris." J Cell Sci. 2010 Dec 1;123(Pt 23):4107-16.

Bermejo C, Haerizadeh F, Takanaga H, Chermak D, Frommer WB. "Dynamic analysis of cytosolic glucose and ATP levels in yeast with optical sensors." Biochem J. 2010 Sep 20

Dechant R, Binda M, Lee SS, Pelet S, Winderickx J, Peter M. "Cytosolic pH is a second messenger for glucose and regulates the PKA pathway through V-ATPase." EMBO J. 2010 Aug 4;29(15):2515-26.

Manzoni R, Montani F, Visintin C, Caudron F, Ciliberto A, Visintin R. "Oscillations in Cdc14 release and sequestration reveal a circuit underlying mitotic exit." J Cell Biol. 2010 Jul 26: 209-22.

Furuya K, Niki H. "The DNA damage checkpoint regulates a transition between yeast and hyphal growth in Schizosaccharomyces japonicus." Mol Cell Biol. 2010 Jun;30(12):2909-17.

Octavio LM, Gedeon K, Maheshri N. "Epigenetic and conventional regulation is distributed among activators of FLO11 allowing tuning of population-level heterogeneity in its expression." PLoS Genet. 2009 Oct;5(10):e1000673.

Thorn K. "Spinning-disc confocal microscopy of yeast." Methods of Enzymology, vol 470, 2010, 581-602.

Lee PJ, Helman NC, Lim WA, Hung PJ. "A microfluidic system for dynamic yeast cell imaging." Biotechniques. 2008 Jan;44(1):91-5.


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