Statement of Work.pdf

Statement of Work Project: Acquire a system that will be able to provide enumeration of viable cells and enumeration of specific single cell populations based on fluorescence labeling

GENERAL INFORMATION

1.0 Scope of Work:

Multiple projects within the Unit require the ability to rapidly enumerate viable eukaryotic cells from cellular debris and non-nucleated cells. In addition, some studies require enumeration of viable cells labeled also with a specific fluorescent tag (to enumerate specific leukocyte populations). Cells from multiple species need enumeration for proper subsequent processing (bovine, swine, sheep, rodents, cervids). The ability to rapidly determine sample concentration, total cell count, and viability are critical steps for downstream applications of multiple project plans goals and can serve as a first quality control point to minimize time and monetary waste on poor quality samples.

2.0 Background:

Require a system that can enumerate eukaryotic cells in a suspension created from various tissues and animal species for downstream assays such as cell culture, metabolic assays, PCR, RNA sequencing, cryopreservation, etc. The system must determine the number of viable cells in a sample in minimal time and with limited input and interpretation from the operator (high level of automation). Some cell enumeration/viability instruments or methods require operator identification of live and total cells with laborious calculations required to determine percent viability and concentration. Other non-laser fluorescence-based methods have trouble identifying small cells and often mislabel small cells as debris or dead cells.

Additionally, large footprint instruments minimize the usefulness of the cell counter when working with biological hazardous samples that require cell counting in a biological safety cabinet. Thus, an instrument that is small, portable, capable of accurately counting a wide range of live eukaryotic cells with minimal sample and operator input is required. In the future, a fluorescent based system may be used to rapidly identify sample purity (percent positive for individual cell populations) and viability to select highest quality samples for downstream assays such as single-cell RNA sequencing, bulk RNA sequencing, metabolic assays such as Seahorse.

CONTRACTOR REQUIREMENTS

3.0 Technical Requirements/Tasks:

1) Instrument must use laser-based fluorescence detection via microcapillary system for detection and enumeration of cells.

2) Instrument must allow for detection of multiple fluorescent colors (minimum 2) as well as forward scatter.

3) Instrument must have rapid processing time once sample is loaded into instrument (< 1 minute per sample is preferred; up to 2 min max)

4) Instrument must report total cells in sample, percent live cells, total live cells, live cells/ml, and total cells/ml with minimal input from operator.

5) Instrument must be small and portable to allow use in a biosafety cabinet (< 15lb, fit in BSC (max of 15in x 15in x 15 in).

6) Instrument must require minimal set up to allow movement within lab and across laboratories as needs arise across unit.

7) Computer system must be built into the instrument to minimize footprint on bench and enhance ability to decontaminate instrument when used in biosafety cabinet

8) Assays should require minimal sample volume (approximately 200ul)

9) Data files must be output in Flow Cytometry Standard (FCS) 3.0 and comma-separated value (CSV) format at a minimum, but instrument should also display results in real time either in summary format or in representative dot plots.

10) Instrument should have capabilities of detect eukaryotic cells of multiple sizes (approximately 2-60 um in diameter). Ability to enumerate bacteria is not required.

11) Unit shall utilize one-step reagents to label cells for enumeration of viable cells.

12) Qty: 1 unit.

4.0 Deliverables/Schedule

Shall be delivered onsite within 45 days of date of award.

5.0 Evaluation Factors

Lowest price technically acceptable and delivery ability

6.0 Place of Performance: USDA, 1920 Dayton Ave, Ames, IA 50010

7.0 Technical Point of Contact: Should there be technical questions, please contact

Jayne Wiarda at 515-337-6353 or jayne.wiarda@usda.gov.