Source: ARRYX, INC. submitted to NRP
HOLOGRAPHIC OPTICAL CELL MANIPULATION FOR AUTOMATED DEVELOPMENT OF IMPROVED YEAST STRAINS FOR ETHANOL PRODUCTION
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
SMALL BUSINESS GRANT
Reporting Frequency
Annual
Accession No.
0206608
Grant No.
2006-33610-16796
Cumulative Award Amt.
$79,998.00
Proposal No.
2006-00483
Multistate No.
(N/A)
Project Start Date
May 1, 2006
Project End Date
Dec 31, 2007
Grant Year
2006
Program Code
[8.8]- (N/A)
Recipient Organization
ARRYX, INC.
316 NORTH MICHIGAN AVENUE
CHICAGO,IL 60601
Performing Department
(N/A)
Non Technical Summary
Potentially insufficient corn stock supplies make necessary the development of industrial yeast strains for converting alternative cellulosic materials for renewable fuel sources. This project investigates the feasibility and develops the capability of using Holographic Optical Trapping as an automated manipulation tool for yeast cells, to characterize and evaluate yeast strains.
Animal Health Component
20%
Research Effort Categories
Basic
20%
Applied
20%
Developmental
60%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
1311799110310%
1314020104010%
5111699108010%
5111799110310%
5114020100010%
5114020104010%
5115310202010%
5117299103010%
5117299108010%
5117410103010%
Knowledge Area
511 - New and Improved Non-Food Products and Processes; 131 - Alternative Uses of Land;

Subject Of Investigation
1799 - Fiber crops, general/other; 1699 - Pasture and forage crops, general/other; 4020 - Fungi; 5310 - Machinery and equipment; 7299 - Research equipment and methods, general/other; 7410 - General technology;

Field Of Science
1040 - Molecular biology; 2020 - Engineering; 1103 - Other microbiology; 1030 - Cellular biology; 1000 - Biochemistry and biophysics; 1080 - Genetics;
Goals / Objectives
The overall objective of this program is to demonstrate the feasibility of manipulating cells within the wells of a standard 96-well micro plate by means of HOT technology. This manipulation of individual and groups of cells would then become part of a larger effort to automate the experimental development process of functional recombinant protein expression from full-length cDNA libraries.
Project Methods
The BioRyx 200 system will be adapted to work with a 96-well plate, and a procedure and associated software will be developed for stage control to access a given well. A procedure and associated software will be implemented to estimate yeast cell growth within each well. The procedure and software for capturing cells within each well and obtaining fluorescence information will be developed. This will include rotation of cells for multi-perspective imaging.

Progress 05/01/06 to 04/30/07

Outputs
OUTPUTS: The BioRyx 200(Arryx's Holographic Optical Trapping (HOT) system) was appropriately modified to work with flat-bottomed 96-well plates and allow optical trapping of yeast cells suspended in solution for analysis. Protocols for growth of the various yeast strains and their subsequent processing and imaging were developed. High magnification brightfield images provided information about the growth, morphology and shape of the different yeast strains, while fluorescence images provided details about the target protein expression. PARTICIPANTS: Arryx Investigators: 1 - Nicole Grynaviski (PI) 2 - Evan Tanner 3 - Jessica Barr 4 - Tania Chakrabarty 5 - Dan Mueth Collaborators: Stephen Hughes, ncaur - usda PROJECT MODIFICATIONS: AS previously reported: 1) Change of Legal Status - Arryx Inc was acquired by Haemonetics effective July 18th 2006 and is now a wholly-owned subsidiary. 2) Change in Project Leadership - The original PI/PD: Joseph Plewa has left the company and Arryx Inc needs approval of the replacement PI/PD, Nicole Grynaviski. Her signature signifying willingness to assume leadership, a copy of her c.v. and updated primary contact forms are included.

Impacts
Significant advancements in software development were required to allow for the possibility of automated collection and analysis compatible with the automated workcell platform. It was found that HOT can be used to trap multiple cells simultaneously and yeast cells are easily rotated along the long axis (particularly if any budding is present) to achieve different orientations.

Publications

  • No publications reported this period