Recent Technologies on 2D and 3D Imaging Flow Cytometry.

Imaging flow cytometry is a technology that performs microscopy image analysis of cells within flow cytometry and allows high-throughput, high-content cell analysis based on their intracellular molecular distribution and/or cellular morphology. While the technology has been available for a couple of decades, it has recently gained significant attention as technical limitations for higher throughput, sorting capability, and additional imaging dimensions have been overcome with various approaches. These evolutions have enabled imaging flow cytometry to offer a variety of solutions for life science and medicine that are not possible with conventional flow cytometry or microscopy-based screening.

High-speed 3D imaging flow cytometry with optofluidic spatial transformation.

Three-dimensional (3D) fluorescence imaging is important to accurately capture and understand biological structures and phenomena. However, because of its slow acquisition speed, it was difficult to implement 3D fluorescence imaging for imaging flow cytometry. Especially, modern flow cytometers operate at a flow velocity of 1-10 m/s, and no 3D fluorescence imaging technique was able to capture cells at such high velocity.

In silico-labeled ghost cytometry.

Characterization and isolation of a large population of cells are indispensable procedures in biological sciences. Flow cytometry is one of the standards that offers a method to characterize and isolate cells at high throughput. When performing flow cytometry, cells are molecularly stained with fluorescent labels to adopt biomolecular specificity which is essential for characterizing cells.

Reduced acoustic resonator dimensions improve focusing efficiency of bacteria and submicron particles.

In this study, we demonstrate an acoustofluidic device that enables single-file focusing of submicron particles and bacteria using a two-dimensional (2D) acoustic standing wave. The device consists of a 100 μm × 100 μm square channel that supports 2D particle focusing in the channel center at an actuation frequency of 7.39 MHz.

Ghost cytometry.

Ghost imaging is a technique used to produce an object's image without using a spatially resolving detector. Here we develop a technique we term "ghost cytometry," an image-free ultrafast fluorescence "imaging" cytometry based on a single-pixel detector. Spatial information obtained from the motion of cells relative to a static randomly patterned optical structure is compressively converted into signals that arrive sequentially at a single-pixel detector.

High-throughput label-free image cytometry and image-based classification of live Euglena gracilis.

We demonstrate high-throughput label-free single-cell image cytometry and image-based classification of Euglena gracilis (a microalgal species) under different culture conditions. We perform it with our high-throughput optofluidic image cytometer composed of a time-stretch microscope with 780-nm resolution and 75-Hz line rate, and an inertial-focusing microfluidic device. By analyzing a large number of single-cell images from the image cytometer, we identify differences in morphological and intracellular phenotypes between E.

High-throughput optofluidic particle profiling with morphological and chemical specificity.

We present a method for high-throughput optofluidic particle analysis that provides both the morphological and chemical profiles of individual particles in a large heterogeneous population. This method is based on an integration of a time-stretch optical microscope with a submicrometer spatial resolution of 780 nm and a three-color fluorescence analyzer on top of an inertial-focusing microfluidic device. The integrated system can perform image- and fluorescence-based screening of particles with a high throughput of 10,000 particles/s, exceeding previously demonstrated imaging particle analyzers in terms of specificity without sacrificing throughput.