2_JA 2084127 Nexcelom Celigo.docx

Justification for Other than Full and Open Competition:

at or Below the Simplified Acquisition Threshold

Project Title: Celigo Image Cytometer

I. Identification of the agency and contracting activity

II. DHHS, NIH, NIAID, LAD

Address of the requesting lab/office: 10 Center Drive, Bethesda Maryland, 20892

III. Description of the project, and supplies or services required to meet the agency’s needs.

A. Project Background

The Laboratory of Allergic Disease(LAD) conducts basic and clinical research on the function of the immune system with emphasis on immunological diseases. The studies are aimed at understanding the physiological control of lymphocyte proliferation, survival, signal transduction pathways in inflammation and understanding pathogenesis of allergic inflammation; of induction of lymphocyte effector functions in tissues; and to identify novel immunomodulatory and anti-inflammatory approaches to the treatment of allergic and immunologic disorders. The information gained by these studies will benefit public health by providing a better understanding of how lymphocytes respond to foreign substances, and as to how adverse responses to self, such as auto-immune diseases, are avoided. Such an understanding will be of great importance in treatment strategies for various allergic diseases and autoimmune diseases. Celigo Imaging Cytometer is invaluable in this effort as it can simultaneously provide data on many of the cell surface markers and protein expression and to elucidate lymphoid cells involvement in allergic responses. This instrument will be heavily used and are the essential devices to the ongoing research efforts of the NIAID. It will markedly improve research data collection.

B. Description of the supplies/services to be purchased

The Celigo Imaging Cytometer is a high-speed, fully automated cytometry system that provides high-throughput, whole-well imaging and quantitative data through image analysis in bright field and four fluorescent channels and 3D imaging. Measurements are compatible with 6 to 1536-well plates and T-flask formats over the course of several days or weeks, or at a single-time measurement. It can be used in adherent and suspension cultured cells, 3D tumor spheroids and colonies of iPSC and cancer stem cells. This instrument will be an essential resource for investigators in Laboratory of Allergic Diseases.

The instrument is used for a wide variety of cell-based assays:

· Apoptosis and cell cycle analysis

· Confluence-based cell growth tracking

· Multi-drug growth curve inhibition

· Migration and invasion assays. In Boyden chambers, Celigo automatically images and counts the migrated cells to the bottom of the well and cells migrated through the filter and adhered to the bottom side of the filter

· Cellular assays for receptor internalization

· Wound healing assays

· Protein expression and detection

· Phosphorylation and phagocytosis

· Killing assays

· Characterization and quantification of entire 3D tumor spheroids or of organoids Dimensions: 19 inch wide x 29 inch long x 17 inch high (with plate holder extended)

IV. Identification of the statutory authority permitting other than full and open competition

☒ 41 USC 1901(a)(1) and FAR Part 13.106-3: Brand Name

V. Demonstration that the proposed contractor(s) unique qualifications or the nature of the acquisition requires use of the authority cited.

A. Describe why only the requested brand or brand name feature of the supply/service will meet the government’s needs

Studies on basic biological processes in mast cells is critical for the understanding of mast cell related diseases such as allergy and mast cell proliferative disorders, and the development of drugs that may reduce their numbers or activity. A limitation in most of our current studies has been the number of cells required. The Celigo imager would allow us to obtain high throughput data with limiting numbers of mast cells to streamline studies on proliferation and death in neoplastic mast cells and mast cells derived from patients under various conditions or drugs, to test the metastatic potential of neoplastic mast cells, their chemotactic behavior and other aspects of mast cell biology. Because of the versatility of the instrument and applicability to many cell-based assays including those involving lymphocyte function and organoids, it would become an essential resource for the Laboratory of Allergic Diseases as a whole.

Celigo offers several advantages to other products on the market which is essential for our research:

· Best-in-class to visualize cells in bright field as some of the mast cells in our study will not be labeled with a fluorochrome. Celigo Imager and software will accurately identify and segment cells in brightfield, other imagers will struggle to complete this task.

· Lab experiments looking at whole well imaging of migration of cells will need to pick up small populations of cells scattered throughout the well that had migrated from the top insert. Celigo Imager is well suited to look at the cells closer to the edges of the well, while other imager will only take a snapshot in the center of the well, missing cells of importance especially for larger 24 well format plates.

· Celigo will reduce experimental time and increase throughput data collection. Unique optics (F-theta lens) allows for very fast whole plate scan time.

· Software is easy to use, has a quick learning curve for new members of the lab thus decreased training time, and is fast to process data. Can scan and analyzing data in one setting to save time. Using the experiment/project mode, data is collected in just one/two clicks.

Based on research needs stated above, the following Celigo features are a must:

· utilizes a proprietary optical system consisting of an F-Theta lens and galvanometric mirrors that enable rapid whole well imaging of all SBS format multi-well plates at 1 um/pixel resolution, allows off-axis scanning which can image 1536-well plates for 1 channel in less than 3 min and 4 channels in less than 8 min

· capable of imaging and analyzing whole wells of an entire 96 well plate in two fluorescent colors in less than 9 minutes at 1 um/pixel resolution. Direct image and count suspension and adherent cells in bright fields for whole well of 96-well palters in less than 5 minutes.

· provides best in class bright field imaging with whole well coverage, uniformity of illumination and even contrast to well edge for all SBS format multi-well plates.

· provides noninvasive in situ analysis of adherent and suspension Other Important Celigo Image Cytometer features needed for our studies:

· The Celigo also can perform rapid imaging and analysis of 3D spheroids in a 384-well plate in less than 2 min/plate

· The Celigo can directly count suspension cells and measure viability in a 96-well plate in less than 10 min/plate for cell counts ranging from 30 – 100,000 cells/well

· Count fluorescently stained adherent cells in large format plates (up to 400,000 cells/well in a 6-well plate) in less than 5 min/well

· A combination of bright field and 4 fluorescent channels for imaging up to 5 channels suitable for tracking and quantifying up to 5 different cell populations in a co-culture experiment using suspension and adherent cells

· Ability to image analyze 6- to 1536-well microplates, 10 cm dish, T75 and T25 flasks, and microscope slides

· Able to acquire high quality images with uniform illumination and consistent image contrast from well edge to well edge allowing accurate cellular quantification and characterization for cell-based assays at a resolution of 1 um/pixel

· Specially designed optics with large depth of focus enabled quantification of large 3D spheroids (up to 900 μm diameter) using single image/well instead of a stack of images

· A suite of ready-to-use bright field and fluorescent applications relevant to cell-based assays development

· Bright field applications include cell counting, confluence measurements, growth curves and proliferation, cell morphology analysis, cytopathic effects, cell culture QC, cell line development, colony formation and colony growth monitoring, 3D spheroid analysis for tumor spheres and embryoid bodies, cell migration and wound healing

· Fluorescent applications include cell health, cell viability, apoptosis, DNA synthesis and proliferation, cell cycle, fluorescent marker analysis, transfection and transduction efficiencies, multi-color fluorescent colony analysis and 3D spheroids fluorescent analysis

· Able to track the growth of individual 3D spheroids in a well containing multiple spheroids

· Directly count cells on transwell membranes for cell migration assays in bright field or fluorescence

· The ability to provide measurements for adherent and suspension cells without the need for trypsinization or sample preparation

· Validated over 100 different cell lines for direct cell counting in bright field

· Flexibility of utilization with multiple imaging modes (full well or partial well) and several focusing mechanisms

· Able to quickly review image results from plate-view level to cell-view level with a single magnification to rapidly assess the experimental results

· An easy-to-use user interface for everyday use by laboratory personnel that does not require an image analysis expert

· A flexible data management system supported by a database hosted either locally or within a networked server infrastructure

· Directly export segmented image data to .csv, .fcs, .ice, and .acs file format for flow or image-based analysis

· The ability to be integrated within an automation platform for the development of automated workflows