The article describes the advanced features of the μManager software, an open-source, cross-platform application for controlling microscope hardware. μManager supports a wide range of scientific equipment, including microscopes, cameras, stages, and illuminators. It provides a user-friendly graphical interface and open programming interfaces for plugins and scripts. The software has been widely adopted by researchers worldwide and is used in over 500 publications. The article presents a guide to using recent advanced features of μManager, including hardware synchronization, multi-camera imaging, projection of patterned light, live slide mapping, imaging with multi-well plates, particle localization and tracking, and high-speed imaging. It also describes the High-Content Screening (HCS) plugin, which enables automated acquisition of large datasets using multi-well plates. The plugin allows users to generate XY positions for imaging across multiple wells, and provides an efficient way to prepare a list of positions for multi-dimensional acquisition. The article also discusses high-speed imaging with sCMOS cameras, which can capture images at high frame rates. It provides a guide on setting up a computer for high-speed acquisition, including hardware and software requirements. The Slide Explorer plugin allows users to acquire large areas of a microscope slide that exceed the camera's field of view, by tiling images and synthesizing a mosaic image. The Pixel Calibrator plugin is used to calibrate the camera's pixel coordinates to the XY stage's coordinate system. The article also describes the Localization Microscopy plugin, which automates the analysis of images of single molecule fluorophores, quantum dots, or other localizable nano-scale particles. It provides an advanced user interface for filtering, inspecting, and manipulating localization data sets. The plugin uses rapid algorithms to find diffraction-limited spots, perform PSF fitting, track spots across multiple time points, and reconstruct super-resolution images. The article also discusses hardware synchronization, which allows for fast and accurate timing of image acquisition. It describes the setup and use of hardware triggering, including the use of TTL pulses to synchronize the camera with other devices. The article also describes the use of multiple cameras for simultaneous imaging of multiple fluorescence channels, and the use of the Multi Camera virtual camera device to combine images from multiple cameras. Finally, the article describes the Projector plugin, which enables the use of light projection devices to phototarget specimens. It provides a guide on setting up and using the phototargeting functionality, including calibration of the phototargeting device and testing the results of the calibration procedure. The article also includes troubleshooting information for common issues encountered while using the software.The article describes the advanced features of the μManager software, an open-source, cross-platform application for controlling microscope hardware. μManager supports a wide range of scientific equipment, including microscopes, cameras, stages, and illuminators. It provides a user-friendly graphical interface and open programming interfaces for plugins and scripts. The software has been widely adopted by researchers worldwide and is used in over 500 publications. The article presents a guide to using recent advanced features of μManager, including hardware synchronization, multi-camera imaging, projection of patterned light, live slide mapping, imaging with multi-well plates, particle localization and tracking, and high-speed imaging. It also describes the High-Content Screening (HCS) plugin, which enables automated acquisition of large datasets using multi-well plates. The plugin allows users to generate XY positions for imaging across multiple wells, and provides an efficient way to prepare a list of positions for multi-dimensional acquisition. The article also discusses high-speed imaging with sCMOS cameras, which can capture images at high frame rates. It provides a guide on setting up a computer for high-speed acquisition, including hardware and software requirements. The Slide Explorer plugin allows users to acquire large areas of a microscope slide that exceed the camera's field of view, by tiling images and synthesizing a mosaic image. The Pixel Calibrator plugin is used to calibrate the camera's pixel coordinates to the XY stage's coordinate system. The article also describes the Localization Microscopy plugin, which automates the analysis of images of single molecule fluorophores, quantum dots, or other localizable nano-scale particles. It provides an advanced user interface for filtering, inspecting, and manipulating localization data sets. The plugin uses rapid algorithms to find diffraction-limited spots, perform PSF fitting, track spots across multiple time points, and reconstruct super-resolution images. The article also discusses hardware synchronization, which allows for fast and accurate timing of image acquisition. It describes the setup and use of hardware triggering, including the use of TTL pulses to synchronize the camera with other devices. The article also describes the use of multiple cameras for simultaneous imaging of multiple fluorescence channels, and the use of the Multi Camera virtual camera device to combine images from multiple cameras. Finally, the article describes the Projector plugin, which enables the use of light projection devices to phototarget specimens. It provides a guide on setting up and using the phototargeting functionality, including calibration of the phototargeting device and testing the results of the calibration procedure. The article also includes troubleshooting information for common issues encountered while using the software.