Multiphoton scanning flow cytometer for multicellular aggregates

摘要

A flow cytometry system suitable for characterizing multicellular aggregates during culture and before implantation combines a low shear flow channel with a multiphoton laser scanning microscope, the latter permitting the characterization of interior and exterior cells in optical isolation from other cells for a representative sampling of fluorescent activity. Imaging capabilities permit sophisticated statistical measurements reflecting individual cell characteristics.

主权项

1. A flow cytometry system comprising:
a pump providing an unconfined volume of liquid subject to an applied force; a channel receiving a liquid flow from the pump, the channel containing the liquid flow along an axis providing a hydrodynamic focusing of a multicellular aggregate within the liquid without disruption of intercellular connections of the multicellular aggregate; a multiphoton laser scanning microscope positioned to illuminate multicellular aggregates within the liquid flow and to record a fluorescence of multiple cells of the multicellular aggregates isolated to a focal plane through the multicellular aggregates as the multicellular aggregates reach an analysis point in the channel; and a control system executing at least one stored program to receive data from the multiphoton laser scanning microscope to provide an output signal providing an assessment of the multicellular aggregates based on recorded fluorescence from the multiple cells along the focal plane.