Specifications_Cell Sorter.pdf

SPECIFICATION

FACS Cell Sorter

I BACKGROUND

The USDA-ARS, Grape Genetics Research Unit (GGRU) in Geneva, NY aims to lead an effort in the development of technologies for the generation of transgene-free genome edited grapes as well as the breeding and production of elite wine and table varieties. Our research group requires the purchase of a cell sorter/flow cytometry unit for Fluorescent Activated Cell Sorting (FACS) analysis. Currently, the GGRU is reliant on outsourcing our cell sorting needs from either local cell sorting facilities in New York, or out of state at locations that specialize in plant cell sorting. Specifically, there are two main needs for a cell sorter at our location.

1. The enrichment of successfully transfected grape protoplasts for the development of transgene-free, genome edited grapevine.

2. Ploidy analysis for the evaluation of plant germplasm at our location.

Currently our research group has developed protocols for the isolation, transfection, and regeneration of grapevine protoplasts from embryogenic callus. Due to the nature of embryonic callus, the transfection efficiency is very low, approximately between 1-10%. We would like to purchase a cell sorter capable of FACS analysis to aid in the sorting and enrichment of our successfully transfected protoplasts for the regeneration of edited grapevine. The enrichment process will not only ensure we are regenerating successfully transfected protoplasts, but, it will also drastically reduce the amount of time, labor, and material resources, put into the genotyping of the regenerated population, as well as, limiting the possibility of contamination of our samples compared to outsourcing the cell sorting service.

This cell sorter would be critical for our mission to develop transgene-free genome edited grapevine to aid in sorting and enrichment of our transfected protoplasts. We are looking to obtain a cell sorter with a small footprint that is capable of fluorescence analysis, easy to use with minimal upkeep, and finally, can be used in a biosafety cabinet with disposable cartridges ensuring a sterile environment for our sorted protoplast. This cell sorter would also be able to meet our second need, to aid in the ploidy analysis of the germplasm managed by the breeders at our location.

II. SYSTEM REQUIREMENTS.

1. Cell Sorter

a. The cell sorter must be able to sort viable plant protoplasts as demonstrated by in-person demo with our samples.

b. The cell sorter must be able to sort cells in custom buffers that have high osmolarity greater than 490m Osm/L.

c. The cell sorter must have fluorescent capabilities for Green Fluorescent Protein (GFP) Propidium Iodide (PI) and 4′,6-diamidino-2-phenylindole (DAPI) nuclear stain.

d. The cell sorter must have a small footprint and compatible to be operated within a biosafety cabinet.

The maximum dimensions of the cell sorter should 15” x 18” x 14” (HxLxW) and under 60lbs.

e. The cell sorter must have disposable cartridges and microfluidics lines to ensure sterility between the transfection and the regeneration process.

f. The cell sorter must have a low nozzle pressure, 2 psi or less to minimize shearing forces exerted on protoplasts after transfection.

g. The cell sorter must have a large 70 x 70 micron sorting channel.

h. The cell sorter must be able to perform absolute counts.

i. The cell sorter must be able to have up to 95% sorting purity.

j. The cell sorter must have an analysis speed of at minimum 5000 events/second.

k. The cell sorter sample input must be compatible with 1.5mL and 5mL tubes.

l. The cell sorter sample output much be compatible with 1.5mL or 5mL tubes.

m. The cell sorter must have a stirrer to gently agitate the protoplasts due to longer sort with low nozzle pressure.

n. The cell sorter must have a temperature-controlled sample and sheath buffer.

o. The cell sorter must have 2-way sorting to sort for multiple populations among a single bulk sort.

p. The cell sorter must include software and training for the software.

q. The cell sorter must include a computer for the software.

r. The cell sorter must come with calibration beads.

s. The sell sorter must include a minimum 1-year warranty covering parts and remote technical services.

t. The cell sorter must include installation and on-site training services.

III. Section 508 Accessibility Compliance

It is mandatory that products developed, procured, maintained or use Electronic and Information Technology (EIT) be accessible to employees with disabilities. Examples of EIT equipment are peripherals, copiers, fax machines, computers.

All Electronic and Information Technology (EIT) procured through this Contract must meet the applicable accessibility standards at 36 CFR 1194, unless an agency exception to this requirement exists.

(36 CFR 1194 implements Section 508 of the Rehabilitation Act of 1973, as amended, and are viewable at: http://www.section508.gov/, then go to 508 Law.

IV. Delivery and System Installation:

a. System Footprint. Not applicable.

b. Government Furnished Property. Not applicable

c. Utilities. Not applicable.

d. Delivery. The Contractor shall furnish and deliver (F.O.B. Destination, Within Consignee’s

Premises) the equipment and documentation to the USDA, ARS, NAA, Grape Genetic Resources Unit, 630 W. North St., Bldg. 4974, Geneva, NY, 14456, within sixty (60) days after the receipt of the purchase order. Delivery shall be between the hours of 8:00 a.m. to 4:30 p.m., Monday through Friday, excl. Federal holidays. The Contractor shall provide the addressee advance notification of delivery.

Such notice shall be provided in writing, not less than two (2) weeks prior to delivery. The Contractor shall provide at least 24 hours advance notice to the requester prior to actual delivery. Collect calls will not be accepted.

e. Installation and Start-up.

V. Training and Documentation

VI. Warranty. The Contractor shall provide standard warranty coverage.

http://www.section508.gov/