Design, Develop, Deliver: 10 FAQs About PhenoVista Biosciences’ Assay Development Services

As one of the leading providers of custom imaging-based phenotypic assays, PhenoVista Biosciences receives all types of inquiries regarding their assay development services. Here are 10 of the most frequent ones answered by Vicki Racicot, Director of Application Scientists, and Matt Stoltz, Sr. Director of Business Development.

What are typical types of high-content, cell-based assays that you offer? Are these standardized workflows, or is customization possible?
PhenoVista’s core capabilities focus on the development, execution and analysis of cell-based assays through high-content imaging across numerous areas of biology/disease. We routinely design and develop customized workflows to generate diverse readouts at the cellular and subcellular levels – from whole-cell marker positivity to nuclear/organellar localization and beyond.

Client-project designs range from completely bespoke, development projects to straightforward onboarding and analysis of client-generated samples.

While we do plan to launch more standardized offerings soon, we currently tailor most projects and associated workflows on a per-project basis. Some general, commonly requested assays include evaluation of mitochondrial or cellular health, uptake and localization of test articles and detection and co-localization analysis of markers of interest.

What type of markers and readouts can you accommodate in a typical study scope? Is there a limit to the total number of markers that you can assess in each study?
Theoretically we can accommodate assessment of any number of markers, provided that robust detection reagents are available that address the relevant biology and are compatible with PhenoVista’s in-house, high-content imaging platforms. We define “palettes” of up to four compatible markers per grouping, tailored to ensure cross-compatibility in the staining workflow and to minimize potential spectral crosstalk between fluorophores. Each project may incorporate use of one or more palettes to capture all desired measurements.

What types of 3D assays do you offer? Which cell types can you integrate into these models?
PhenoVista has extensive expertise in developing and working with 3D, in vitro cell models including spheroids, organoids and 3D, printed co-cultures. We can incorporate a wide array of cell types into these models. These include, but are not limited to, immortalized cells, primary cells (including immune cells), iPSCs and/or patient-derived cells. We have used these models in diverse assays, such as CAR-T cell-mediated cancer organoid cytotoxicity, synovial-joint modeling and gut neurosphere differentiation.

Can you execute time-resolved studies that require imaging of live cells?
Yes, we can design time-resolved studies based on two different approaches, depending upon the desired data output.

For studies that require fully kinetic readouts, we can utilize live cells that either express fluorescently tagged marker(s) and/or are compatible with fluorescent indicator dyes that do not impact cell viability and image over time in our live cell-enabled imaging instruments (e.g., Incucyte, GE IN Cell Analyzer 6500HS). This approach is best suited to small-scale projects that do not require high-throughput data generation.

If the study design incorporates analysis of markers that require fixation and/or high-throughput assessment, we can set up a multi-timepoint workflow to address this need.

Do your services require transfer of an established assay protocol, or can PhenoVista support assay development?
Protocol transfer is not required, as our assay services offer tailorable, comprehensive solutions that span initial assay development and setup, staining and imaging and data analysis and mining. Clients can choose to collaborate with us at any point in this process. We can certainly onboard customer protocols, fixed-cell plates, stained-cell plates or previously captured images for downstream processing and analysis.

We can also incorporate outbound technology transfer into project plans, should clients prefer to transition any developed assay protocol back to their organization for subsequent in-house use.

What types of cell models can you accommodate in your assay services? Do you offer analysis of tissues?
We can utilize a wide array of cell types in our assay services, including immortalized, primary, iPSCs and patient-derived cells. While we do not offer tissue analysis, PhenoVista has established expansive 3D, high-content imaging assays to characterize more complex, physiologically relevant cellular models, such as spheroids, organoids and 3D, printed co-cultures that incorporate diverse cell types.

Can you accommodate non-imaging readouts (such as genomic characterization or biochemical assays) into a project plan?
PhenoVista’s core expertise is based in high-content imaging and assay development, execution and analysis. However, we do recognize that your project may require generation of complementary data sets to achieve your goal(s). To that end, we can offer parallel generation of cellular samples for external analysis as a deliverable in your study. Where appropriate, we may suggest partnering with one of our collaborators to analyze these samples, and we can incorporate contracting these additional services into your project plan.

What is the typical scale of a high-content screening project? Can your services accommodate large-scale screens, for example, up to thousands of compounds?
The potential throughput of any study will depend on a number of factors, such as assay format and availability of reagents, cell types or other required materials. While most clients engage us to execute smaller-scale, in-depth studies to explore, for example, therapeutic mode-of-action, we can certainly accommodate larger-scale studies to assess more expansive collections of therapeutics.

What therapeutic modalities can you evaluate in a high-content, cell-based assay? Do you maintain an in-house compound library for use in client projects?
Depending on the assay design, we have the flexibility to evaluate a broad spectrum of therapeutic agents, including:

• Small molecules
• Targeted therapeutic antibodies
• Antibody-drug conjugates (ADCs)
• Other large and small biologics (such as natural products and peptides)
• Therapeutic siRNAs
• CAR-T cells

While we may have common control compounds on hand in-house, PhenoVista typically onboards therapeutic test agents from customers and/or commercial sources on a project-by-project basis.

Can you generate induced pluripotent stem cells (iPSCs) for imaging studies?
No, we do not generate iPSCs. However, we routinely source them from various commercial providers. PhenoVista has established a partnership with FUJIFILM Cellular Dynamics, Inc. (FCDI) to utilize their high-quality, reproducible iPSC models for client studies. We are also listed as an FCDI preferred provider of contract, cell-based assay services for North America. Additionally, we can tap into a broader provider network to help clients source the most suitable cell model(s) for their project(s).

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