Microfluidic
Tilt Microscopy
Focus
stacks are useful for a variety of purposes in imaging.
Recently, we have demonstrated that fluid flow can
transport a sample through focus with nanometer
accuracy. Using fluid flow, focus stacks of
individual cells can be produced in a single millisecond
at throughputs of 100's of focus stacks per second.

|
A linear
channel tilted at an angle transports cells through
focus
Collected focus stacks of a red blood cell and a
Leukemia cell
|
One
application of focus stacks is to produce phase images
(using the Transport of Intensity Equation) that quantify
optical path differences across the cell. Phase
images can be used to resolve cellular features that are
not visible in bright-field images or to quantify cell
morphology in depth.

|
Phase image of a red blood
cell in flow taking a "parachute" shape
|
Phase
image of a Leukemia cell in flow, where the
pathlength of sub-cellular features can be
resolved
|
|
S. S. Gorthi and E. Schonbrun, "Phase imaging flow cytometry using a focus-stack
collecting microscope," Opt. Lett, 37, 707-709 (2012). |