Abstract: The present disclosure generally relates to systems and methods for multi-focal imaging, for example, for determining nucleic acids in cells or other samples. In some cases, multiple focal planes may simultaneously be determined, e.g., by using a plurality of detectors, such as a plurality of cameras, which image the same sample, but at least some of which are focused on different focal planes within the sample. Thus, the sample may be imaged in 3 dimensions, e.g., without sample refocusing. In certain cases, this may improve the resolution of imaging, in space and/or time. Various embodiments can be used to increase imaging throughput and/or resolution in image-based approaches, e.g., for single-cell molecular profiling such as multiplexed error robust fluorescence in situ hybridization (MERFISH), or for other applications.

Abstract: The present invention generally relates to super-resolution microscopy. For example, certain aspects of the invention are generally directed to a microscopy system comprising at least two objectives. In some embodiments, the microscopy system may also contain a non-circularly-symmetric lens. One or more images can be obtained using the objectives, for example, using stochastic imaging techniques such as STORM (stochastic optical reconstruction microscopy), optionally in conjunction with entities that are photoactivatable and/or photo switchable. The images obtained using the objectives may be compared, e.g., to remove noise, and/or to compare an entity present in both images, for instance, to determine the z-position of the entity. In some cases, surprisingly high resolutions may be obtained using such techniques, for example, resolutions of better than about 10 nm.

Abstract: A device for confining an object to a region proximate to a fluid flow stagnation point includes one or more inlets for carrying the fluid into the region, one or more outlets for carrying the fluid out of the region, and a controller, in fluidic communication with the inlets and outlets, for adjusting the motion of the fluid to produce a stagnation point in the region, thereby confining the object to the region. Applications include, for example, prolonged observation of the object, manipulation of the object, etc. The device optionally may employ a feedback control mechanism, a sensing apparatus (e.g., for imaging), and a storage medium for storing, and a computer for analyzing and manipulating, data acquired from observing the object. The invention further provides methods of using such a device and system in a number of fields, including biology, chemistry, physics, material science, and medical science.