Optical imaging techniques in microfluidics and their applications

Microfluidic devices have undergone rapid development in recent years and provide a lab-on-a-chip solution for many biomedical and chemical applications. Optical imaging techniques are essential in microfluidics for observing and extracting information from biological or chemical samples. Traditionally, imaging in microfluidics is achieved by bench-top conventional microscopes or other bulky imaging systems. More recently, many novel compact microscopic techniques have been developed to provide a low-cost and portable solution. In this review, we provide an overview of optical imaging techniques used in microfluidics followed with their applications. We first discuss bulky imaging systems including microscopes and interferometer-based techniques, then we focus on compact imaging systems that can be better integrated with microfluidic devices, including digital in-line holography and scanning-based imaging techniques. The applications in biomedicine or chemistry are also discussed along with the specific imaging techniques

Fluorescence microscopy imaging with a Fresnel zone plate array based optofluidic microscope

We report the implementation of an on-chip microscope system, termed fluorescence optofluidic microscope (FOFM), which is capable of fluorescence microscopy imaging of samples in fluid media. The FOFM employs an array of Fresnel zone plates (FZP) to generate an array of focused light spots within a microfluidic channel. As a sample flows through the channel and across the array of focused light spots, the fluorescence emissions are collected by a filter-coated CMOS sensor, which serves as the channel’s floor. The collected data can then be processed to render fluorescence microscopy images at a resolution determined by the focused light spot size (experimentally measured as 0.65 mm FWHM). In our experiments, our established resolution was 1.0 mm due to Nyquist criterion consideration. As a demonstration, we show that such a system can be used to image the cell nuclei stained by Acridine Orange and cytoplasm labeled by Qtracker

The application of Fresnel zone plate based projection in optofluidic microscopy

Optofluidic microscopy (OFM) is a novel technique for low-cost, high-resolution on-chip microscopy imaging. In this paper we report the use of the Fresnel zone plate (FZP) based projection in OFM as a cost-effective and compact means for projecting the transmission through an OFM's aperture array onto a sensor grid. We demonstrate this approach by employing a FZP (diameter = 255 µm, focal length = 800 µm) that has been patterned onto a glass slide to project the transmission from an array of apertures (diameter = 1 µm, separation = 10 µm) onto a CMOS sensor. We are able to resolve the contributions from 44 apertures on the sensor under the illumination from a HeNe laser (wavelength = 633 nm). The imaging quality of the FZP determines the effective field-of-view (related to the number of resolvable transmissions from apertures) but not the image resolution of such an OFM system -- a key distinction from conventional microscope systems. We demonstrate the capability of the integrated system by flowing the protist Euglena gracilis across the aperture array microfluidically and performing OFM imaging of the samples

Quantitative surface normal measurement by a wavefront camera

A compact wavefront camera that allows users to quantitatively measure the intensity and wavefront at a remote object plane is reported. The camera is built from a chip-scale wavefront sensor that we previously developed. By measuring the wavefront of the image and calibrating the wavefront relationship between the image and object planes, the wavefront at the object plane can be computed and the surface normal of the object can be derived. We built a prototype camera and calibrated the wavefront relationship. In a proof-of-concept experiment, a set of concave mirrors with different focal lengths (50–200 mm), were imaged. The results agree well with their expected values. To demonstrate the application of the camera, we applied this method to measure the deformation of a microfluidic channel under pressure

A wide field-of-view microscope based on holographic focus grid

We have developed a novel microscope technique that can achieve wide field-of-view (FOV) imaging and yet possess resolution that is comparable to conventional microscope. The principle of wide FOV microscope system breaks the link between resolution and FOV magnitude of traditional microscopes. Furthermore, by eliminating bulky optical elements from its design and utilizing holographic optical elements, the wide FOV microscope system is more cost-effective. In our system, a hologram was made to focus incoming collimated beam into a focus grid. The sample is put in the focal plane and the transmissions of the focuses are detected by an imaging sensor. By scanning the incident angle of the incoming beam, the focus grid will scan across the sample and the time-varying transmission can be detected. We can then reconstruct the transmission image of the sample. The resolution of microscopic image is limited by the size of the focus formed by the hologram. The scanning area of each focus spot is determined by the separation of the focus spots and can be made small for fast imaging speed. We have fabricated a prototype system with a 2.4-mm FOV and 1-μm resolution. The prototype system was used to image onion skin cells for a demonstration. The preliminary experiments prove the feasibility of the wide FOV microscope technique, and the possibility of a wider FOV system with better resolution

Full field phase imaging using a harmonically matched diffraction grating pair based homodyne quadrature interferometer

In this letter, the authors present a novel quadrature interferometry method based on the use of a harmonically matched shallow grating pair. Unlike a simple beam splitter or single shallow grating, the grating pair can confer a nontrivial interference phase shift (other than 0° or 180°) between the output ports of the interferometer. Using the grating pair as the beam splitter/combiner, the authors implement a homodyne quadrature full field phase interferometer and demonstrate the system's capability to acquire phase and amplitude images

Wide field-of-view microscope based on holographic focus grid illumination

We have developed a new microscopy design that can achieve wide field-of-view (FOV) imaging and yet possesses resolution that is comparable to a conventional microscope. In our design, the sample is illuminated by a holographically projected light-spot grid. We acquire images by translating the sample across the grid and detecting the transmissions. We have built a prototype system with an FOV of 6mm×5mm and acquisition time of 2.5s. The resolution is fundamentally limited by the spot size—our demonstrated average FWHM spot diameter was 0.74μm. We demonstrate the prototype by imaging a U.S. Air Force target and a lily anther. This technology is scalable and represents a cost-effective way to implement wide FOV microscopy system

The optofluidic microscope - A high resolution microscope-on-a-chip system

In this article, a complete on-chip high resolution OFM device is presented. The OFM method abandons the conventional microscope design and, instead, shares some similarities with direct shadow imaging method where the target object is directly placed on top of a CCD or CMOS imaging sensor chip. In direct shadow imaging, the image resolution is no better than the sensor pixel size. As the typical sensor pixel size is around 10 microns, direct shadow imaging systems are unable to provide images that are comparable in resolution to those of conventional microscopes

The application of on-chip optofluidic microscopy for imaging Giardia lamblia trophozoites and cysts

The optofluidic microscope (OFM) is a lensless, low-cost and highly compact on-chip device that can enable high-resolution microscopy imaging. The OFM performs imaging by flowing/scanning the target objects across a slanted hole array; by measuring the time-varying light transmission changes through the holes, we can then render images of the target objects at a resolution that is comparable to the holes’ size. This paper reports the adaptation of the OFM for imaging Giardia lamblia trophozoites and cysts, a disease-causing parasite species that is commonly found in poor-quality water sources. We also describe our study of the impact of pressure-based flow and DC electrokinetic-based flow in controlling the flow motion of Giardia cysts—rotation-free translation of the parasite is important for good OFM image acquisition. Finally, we report the successful microscopy imaging of both Giardia trophozoites and cysts with an OFM that has a focal plane resolution of 0.8 microns

Observing dynamics of transparent samples by harmonically matched grating-based full-field quadrature phase interferometer

Our group has reported the use of harmonically matched diffraction grating for full-field quantitative phase imaging. In this paper, we show the improvement of this technique and the application in observing dynamics of transparent samples. By using the grating as a beam splitter/combiner in an interferometer, we are able to obtain non-trivial phase difference between the output ports of the grating. We have built a Mach-Zehnder interferometer using the holographic grating with 600 and 1200 lines/mm spacing. Two CCD cameras at the output ports of the grating-based Mach-Zehnder interferometer are used to record the full-field quadrature interferograms, which are subsequently processed to reconstruct the phase image. Since the two quadrature interferograms are acquired at the same time, the imaging speed of the system is limited only by the frame rate of the CCD cameras. We have demonstrated the capability of our system by observing dynamics of transparent samples