qHCR Flow Cytometry
qHCR flow cytometry enables RNA relative quantitation for high-throughput expression profiling of mammalian and bacterial cells without the need to engineer reporter lines.
Analog Quantitative Signal
qHCR signal is analog in the form of single-cell fluorescence intensities that scale approximately linearly with the number of RNA target molecules per cell.
Quantitative RNA flow cytometry
Mammalian Cells (HEK)
Redundant 2-Channel Detection
To illustrate quantitative accuracy and precision for single-cell measurements with RNA flow cytometry, we detect a target mRNA using two probe sets that trigger different HCR amplifiers carrying spectrally distinct fluorophores.
Bacterial Cells (E. coli)
High Accuracy and Precision
In the resulting scatter plot of fluorescence single-cell intensities, high accuracy corresponds to linearity with zero intercept, and high precision corresponds to tight scatter around the line.
Orthogonal HCR amplifiers enable 1-step signal amplification for up to 5 target RNAs simultaneously. Straightforward multiplexing is achieved using the same 1-day protocol independent of the number of target RNAs. Learn more
Custom Probe Set Design
Custom probe set design is available for any target mRNA in any organism across the tree of life. Learn more
Automatic Background Suppression
Using HCR v3.0 probes and amplifiers, qHCR flow cytometry benefits from automatic background suppression throughout the protocol, dramatically enhancing performance and ease-of-use. Learn more
Precision Increases with Probe Set Size
Automatic background suppression ensures that precision increases with probe set size. As a result, for qHCR flow cytometry, we recommend using 30 or more split-initiator probe pairs per target RNA (maximize probe set size given target length).
Signal-to-Background Increases with Amplification Time
Signal increases with amplification time and background does not, so for qHCR flow cytometry, we recommend overnight amplification to grow long amplification polymers and enhance signal-to-background.