Flow Cytometry FAQ

Flow cytometry is a powerful tool in drug development for several reasons. These include:

1. High throughput analysis allows for the rapid analysis of thousands of cells per second, making it ideal for screening large libraries of compounds.
2. Flow cytometry can measure multiple parameters simultaneously, such as cell size, granularity, and the expression of various markers. This helps in understanding the complex interactions between drugs and cellular targets.
3. Biomarker identification allows for identifying and evaluation of biomarkers, which are crucial for understanding disease mechanisms and the effects of drugs.
4. For pharmacokinetics and pharmacodynamics (PK/PD), Flow cytometry helps in studying the relationship between drug concentration and its effect on cells, providing insights into the drug’s efficacy and safety.
5. Flow cytometry can detect subtle changes in cell populations, helping to identify potential toxic effects of new drugs early in the development process.

Overall, flow cytometry’s ability to provide data on heterogeneous cell populations makes it an invaluable tool in the drug development pipeline.

TD2 operates as a contract research organization (CRO) offering flow cytometry-based assays for nonclinical studies at both Scottsdale and Bothell sites. The Bothell site offers GLP and non-GLP services as well as support for clinical trials.

TD2’s most experienced flow cytometry scientists have 20+ years working in the field. The flow cytometry staff are skilled in assay development/qualification and are at the forefront for determining the criteria for flow cytometry assay and instrument validation.

1. Three BD FACSCanto II systems with two scatter parameters and up to 8 fluorescent channels
2. Four BD FACSCanto systems with two scatter parameters and up to 10 fluorescent channels
3. Two Cytek Aurora spectral flow cytometers with two scatter parameters and up to 26 fluorescent channels
4. One Miltenyi MACS Quant analyzer with two scatter parameters and up to 14 fluorescent channels

TD2 offers an integrated platform for nonclinical and clinical drug development based on expertise in:

1. Immunophenotyping
   1. T-lymphocyte, B-lymphocyte and Natural Killer cells (TBNK)
   2. T Regulatory Cells
   3. T Memory, Naïve and Effector cells
   4. T Cell Activation
   5. B Memory and Plasma cells
   6. Humanized mice panels
   7. Custom panels are also available
2. Intracellular Protein Analysis
   1. Intracellular Cytokines
   2. Phosphorylated Proteins
3. Cellular Functional Assays
   1. NK Cell Function
   2. Basophil Activation Test (BAT)
4. Cell Cycle and Apoptosis
5. Receptor Occupancy
6. PBMC Services
7. Custom flow cytometry panels and assays available upon request

The technology of flow cytometry also provides invaluable support to the preclinical animal models available at the TD2 Scottsdale facility.

Yes, TD2 can develop assays using literature established protocols for panel design, staining procedures and instrument settings. Our scientists have the knowledge and experience to optimize the conditions for our environment and instrumentation.

Specimen Types

1. Blood: Generally collected in K2EDTA (lavender top) or Sodium Heparin (green top) tubes.
2. Bone Marrow: Generally collected in K2EDTA.
3. Tissue: Fresh tissue samples should be kept in a sterile container with a suitable transport medium.
4. Stability tubes or media can be used for increased stability needs.

Collection and Shipping Guidelines

1. Aseptic technique should be used to avoid contamination.
2. Adequate volume of the specimen needs to be collected per tube size. Typically 1-3 mL for blood and bone marrow.
3. Labeling of all specimens with identifying information and collection date/time should be followed.
4. Specimens should be transported at an appropriate temperature with the proper packaging to preserve specimen integrity and containment and are processed within stability guidelines.

At TD2, we pride ourselves on being able to customize our antibody panels to meet our client’s needs, so we have a limited number of pre-qualified/validated antibody panels. If working with an established antibody panel, TD2 scientists follow best practices for the re-qualification/validation once a flow cytometric method has been modified. This includes an assessment of the impact of different types of assay modifications on assay performance. Recommendations regarding which parameters to evaluate depending on the type of modification and the impact of assay modification on the assay's intended use will be determined. Timelines will depend on the extent of modification and how much troubleshooting is required.

Currently, the flow cytometry laboratories at both the Scottsdale and Bothell sites have sufficient capacity to start new projects within 1-2 weeks after contracts are signed, depending on reagent availability without backorders.

Yes, TD2 can provide the raw .fcs files via ShareFile or via client specific portals for clients to perform their own analysis. These files are typically available within one week of sample collection.

Depending on project scope, TD2 can provide the flow cytometry data in MS Excel tables, graphical representations of the data or a flow cytometry report, either a summary report or a more comprehensive report with detailed analysis. General timelines for each are as follows: Data Tables within 1-2 weeks post collection or monthly for clinical flow projects, Graphing within 2-3 weeks post collection, Summary Report within 2-3 weeks post study end, and Comprehensive Report within 3-4 weeks post study end. These target timelines depend on the deliverables requested and can be impacted by the complexity of the flow cytometry services being performed.

TD2 scientists look forward to exploring potential ways to collaborate with our clients for developing novel flow cytometry assays or discovering novel uses for existing flow cytometry technology leading to potential co-authorship on posters, presentations or manuscripts.

TD2 Scottsdale and Bothell sites welcome the opportunity to host our clients to observe sample processing, conduct on-site audits or just to visit as a part of due diligence. Please contact us to set up an agreed upon time.