Attachment 1 - Specifications - Cell Sorter.pdf

Specifications Full-spectrum cell sorter

1.0 Background:

Several research programs here at Dairy Forage Research Center are presently or evolving to perform more sophisticated whole transcriptome RNA sequencing projects.

Due to the objectives of our newly approved CRISs, our research project focus on the nutritional impact on the dairy cattle and the associated microenvironment in the gastrointestinal environment. Specifically, we are interested in investigating how the diverse cell types in the gut respond to the changes in host nutritional treatments.

Within the microenvironment of the gastrointestinal tissue, different cell types have intricate and complex interactions with other cell types, including immune lineages, endothelial cells, fibroblasts, and extracellular matrix. A comprehensive understanding of the physiological response of the gut tissue to various feed treatment requires thorough interrogation of the immune cell types and their interaction. For cattle, currently there is no reliable markers we can use to confidently labels cell identities. A targeted enrichment of pure cell types is critical to build a reliable set of reference markers to begin downstream analysis.

2.0 Objectives:

Using full-spectrum profiling instrument to identify cell populations of interest with high resolution and sensitivity.

3.0 Required Specifications:

The instrument has to have the minimum performance requirements:

1. Detector channels: 67

a. 64 fluorescence channels,

b. three lasers: one blue laser SSC, one violet laser SSC, and one FSC. The violet

SSC provides improved resolution for small particles

2. Full spectrum profiling technology that has the following capabilities:

a. Detect any fluorochrome that is excited by the on-board laser

b. Capture the entire emission spectra starting from the excitation laser wavelength to the near IR wavelength range

3. High sensitivity Coarse Wavelength Division Multiplexing (CWDM) semiconductor detector arrays that use high quantum efficiency avalanche photodiode (APD) detectors for light capture for dyes emitting in the 365-829 nm range.

4. up to five (5) spatially separated lasers for fixed optical assembly with Flat- Top laser beam profiles with narrow beam heights

5. High bandwidth 22-bit electronics that are scalable up to 67 channels with 6.5 log decades of dynamic range.

6. Cell sorting ability:

a. Sort up to 6-ways into 5ml and 1.5ml tubes; and up to 2-ways into

15ml tubes using a 70, 85, 100, or 130 m nozzle

b. sort cell populations 64 levels deep in the gating hierarchy from multiple cell populations into each tube using default sort modes or custom designed sort modes

c. adjustable guided workflow with default sort modes or custom created sort modes.

d. adjustable drop delay

e. Sort cells into 384-, 96-, or custom defined well plate in high throughput mode

f. Sort a defined number of cells into 384-, 96- or custom defined plates

g. No-contact control of the plate sorting puck

h. Simultaneous analysis of and sorting of 45 colors without compromising data quality (OMIP-109: 45-color full spectrum flow cytometry panel)

7. Software has the following features:

a. Has pre-optimized detector settings for standardize setup

b. proportional adjustment of all detector gains and pre-optimized detector settings without the need of adjustments of individual detectors.

c. Live unmixing of data during sample acquisition and sorting

d. Enable analyzing and sorting using conventional compensation of spillover

e. Live autofluorescence extraction from multiple cell types by utilizing each cellular autofluorescence signal as a unique spectral signature

f. Has guided workflows with default sort modes or custom created sort modes

4.0 Deliverables or Delivery Schedule

The contractor performs the delivery and installation of the new instrument without government supervision. The service will ensure the instrument operates at the original, factor level performance. The contractor provides certified training on operating the instrument and certified workflows Shall be delivered and set up no later than 90 days after award.

5.0 Government Furnished: Government will receive the instrument and review the installation work. The facility is located at 1925 Linden Dr. Madison, WI. It is open Monday-Friday, from 8-4PM.