What is Immunophenotyping?
Immunophenotyping is the characterization and classification of cells in a heterogeneous mixture of cells such as blood, synovial fluid, other aqueous liquids, or solid tissues. It uses the expression of specific genes and combinations of genes to categorize cell types based on known functional characteristics.
The Perfect Balance of High Parameter Data & Speed with Reproducibility
High Parameter Data
With up to 5 lasers and 30 parameters, the ZE5 Cell Analyzer can perform immunophenotyping in depth. In this example, we show a 27-color immunophenotyping panel designed using our cutting edge StarBright™ Dyes to characterize the major human leukocyte sub populations. You can also download the full PDF poster and a technical note with methods and additional data.
27-Color Immunophenotyping Panel Including StarBright, UltraViolet, Violet , Blue, Yellow, and Red Dyes
|HLA DP DQ DR
27-Color Multiplex Panel. Red blood cell lysed human peripheral blood was stained with a live/dead dye (propidium iodide, (PI)) and a 26-color antibody panel in FACS Buffer allowing identification of multiple cell lineages and subsets.
Full Gating Strategy
Click each image to see the full gating strategy:
Speed with Reproducibility
The ZE5 Cell Analyzer can acquire complex immunophenotyping data at rates that significantly exceed conventional instruments. Its innovative fluidics and electronics synergize to enable it to process a 96-well plate in less than 15 minutes whilst still being able to collect tens of thousands of events per well. This means that even complex assays such as the one detailed above can be performed at the speed of a screening instrument and without compromising reproducibility.
To demonstrate this, we repeated the 27-color panel shown above using high throughput mode for sample acquisition. The data shows a high level of reproducibility between high throughput runs and is consistent with standard mode runs. Whilst data quality was preserved the run time for each well was reduced from 3 minutes 21 seconds in standard mode, to 8 seconds in high throughput mode.
Example 27-Color Panel Comparison of Standard and High Throughput Data Acquisition
This example shows six replicates of the 27-color panel data shown previously, a single sample was collected in high throughput (n=4) and standard mode (n=2). Each standard mode replicate was collected in 3 minutes and 21 seconds, while each high throughput mode replicate was collected in 8 seconds.
Powerful Instruments with Cutting Edge Dyes
Explore other example immunophenotyping panels analyzed using the ZE5 Cell Analyzer and see how StarBright Dyes can enhance any panel on any instrument. Includes detailed immunophenotyping protocols.
StarBright Dyes: Build Bigger, Better Panels With Superior Dyes Posters
Flow Immunophenotyping Tools & Services
Building new immunophenotyping panels can be challenging. We have developed a range of resources that can help you get the most out of your flow cytometer regardless of manufacturer or configuration.