: Systems like the ImageStream use Charge-Coupled Device (CCD) cameras with Time Delay Integration (TDI) to track cells in flow, preventing motion blur and enabling the detection of dim markers.
: Researchers must set specific software parameters, such as magnification and laser power, to maximize the resolution of the captured images. Common Applications Imaging Flow Cytometry: Methods and Protocols
Imaging Flow Cytometry (IFC) is a hybrid technology that combines the of conventional flow cytometry with the detailed spatial and morphological resolution of digital microscopy. Unlike standard flow cytometry, which provides integrated signal intensity for a whole cell, IFC captures multiple digital images of every individual cell as it passes through the system. Core Principles and Advantages : Systems like the ImageStream use Charge-Coupled Device
: Cells must be in a single-cell suspension to avoid clogging the thin fluidic channels used for imaging. IFC provides data on cell size
: Non-specific binding is prevented using anti-Fc antibodies. Cells are then stained with fluorophore-conjugated antibodies, viability dyes, or nuclear dyes like DAPI or Hoechst to provide spatial landmarks.
: Beyond fluorescence, IFC provides data on cell size, shape, circularity, and internal complexity.
: If internal markers (e.g., nuclear proteins) are the target, cells are fixed with formaldehyde and permeabilized with detergents like Triton X-100.