How do you validate a flow cytometry assay?

Typically, the validation of a flow cytometry panel for GLP or GcLP applications includes test scripts to assay the following characteristics:

1. precision (includes repeatability and reproducibility)
2. analytical sensitivity [limits of blank (LoB), detection (LoD), quantitation]
3. analytical specificity and interference.

What is assay validation?

Assay Validation: Comprehensive experiments that evaluate and document the quantitative performance of an assay, including sensitivity, specificity, accuracy, precision, detection limit, range and limits of quantitation. Accuracy is the agreement between value found and an excepted reference value (3).

What is flow cytometry assay?

Flow cytometry (FC) is a technique used to detect and measure physical and chemical characteristics of a population of cells or particles. The sample is focused to ideally flow one cell at a time through a laser beam, where the light scattered is characteristic to the cells and their components.

What type of assay is flow cytometry?

Flow cytometry is a technology that provides rapid multi-parametric analysis of single cells in solution. Flow cytometers utilize lasers as light sources to produce both scattered and fluorescent light signals that are read by detectors such as photodiodes or photomultiplier tubes.

Why is validation testing important?

Why is Validation Testing Important? Whenever any particular software is tested then the main motive is to check the quality against the defects being found. The developers fix the bugs and the software is rechecked to make sure that absolutely no bugs are left out in that.

What is method validation?

Method validation is the process used to confirm that the analytical procedure employed for a specific test is suitable for its intended use. Results from method validation can be used to judge the quality, reliability and consistency of analytical results; it is an integral part of any good analytical practice.

Why is assay validation important?

Assays must be precise, robust, and specific during use in preclinical studies and clinical trials in order to assure that drug candidates can be accurately evaluated for safety and efficacy. Validation plans assure that an assay will work reliably, even if an assay is run at different sites or by different users.

What is the purpose of assay test?

Assays are scientific test systems developed for the purpose of evaluating the effects of chemical compounds on target biological processes. These can be cellular, molecular or biochemical, with the goal of furthering drug development and toxicity testing.

Why is flow cytometry important?

The reason flow cytometry is so successful with heterogeneous cell populations is that it analyzes cells one at a time. It does this by using the properties of fluid dynamics. Flow cytometry data enables the user to understand each cell type and its properties on a deeper level.

What is SSC in flow cytometry?

In flow cytometry, the light scattered by cells is measured by two optical detectors: forward scatter (FSC) that detects scatter along the path of the laser, and side scatter (SSC) which measures scatter at a ninety-degree angle relative to the laser.

What is FSC and SSC in flow cytometry?

What is QA validation?

Validation is a process of testing and analyzing operations to guarantee the output they produce will consistently fulfill the end user’s needs and provide the intended medical benefit in actual-use conditions. …