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SATA controller. This is a modern type of storage controller for higher hard disk data throughput, to which the virtual hard disks are attached. Initially you will normally have one such virtual disk, but as shown in the previous screenshot, you can have more than one. Each is represented by a disk image file, such as a VDI file in this example.
You can modify these media attachments freely. For example, if you wish to copy some files from another virtual disk that you created, you can connect that disk as a second hard disk, as in the above screenshot. You could also add a second virtual CD/DVD drive, or change where these items are attached. The following options are available:
Ever since the original IBM PC, personal computers have been equipped with one or two serial ports, also called COM ports by DOS and Windows. Serial ports were commonly used with modems, and some computer mice used to be connected to serial ports before USB became commonplace.
While serial ports are no longer as common as they used to be, there are still some important uses left for them. For example, serial ports can be used to set up a primitive network over a null-modem cable, in case Ethernet is not available. Also, serial ports are indispensable for system programmers needing to do kernel debugging, since kernel debugging software usually interacts with developers over a serial port. With virtual serial ports, system programmers can do kernel debugging on a virtual machine instead of needing a real computer to connect to.
If a virtual serial port is enabled, the guest OS sees a standard 16550A compatible UART device. Other UART types can be configured using the VBoxManage modifyvm command. Both receiving and transmitting data is supported. How this virtual serial port is then connected to the host is configurable, and the details depend on your host OS.
Host Device: Connects the virtual serial port to a physical serial port on your host. On a Windows host, this will be a name like COM1. On Linux or Oracle Solaris hosts, it will be a device node like /dev/ttyS0. Oracle VM VirtualBox will then simply redirect all data received from and sent to the virtual serial port to the physical device.
Raw File: Send the virtual serial port output to a file. This option is very useful for capturing diagnostic output from a guest. Any file may be used for this purpose, as long as the user running Oracle VM VirtualBox has sufficient privileges to create and write to the file.
TCP: Useful for forwarding serial traffic over TCP/IP, acting as a server, or it can act as a TCP client connecting to other servers. This option enables a remote machine to directly connect to the guest's serial port using TCP.
Up to four serial ports can be configured per virtual machine, but you can pick any port numbers out of the above. However, serial ports cannot reliably share interrupts. If both ports are to be used at the same time, they must use different interrupt levels, for example COM1 and COM2, but not COM1 and COM3.
Clicking on the + button to the right of the USB Device Filters window creates a new filter. You can give the filter a name, for later reference, and specify the filter criteria. The more criteria you specify, the more precisely devices will be selected. For instance, if you specify only a vendor ID of 046d, all devices produced by Logitech will be available to the guest. If you fill in all fields, on the other hand, the filter will only apply to a particular device model from a particular vendor, and not even to other devices of the same type with a different revision and serial number.
Serial Number. While vendor ID and product ID are quite specific to identify USB devices, if you have two identical devices of the same brand and product line, you will also need their serial numbers to filter them out correctly.
Multicolor fluorescence microscopy has become a key enabling technology in biology by providing the means to spectrally resolve cells, organelles, or molecules within tissues and to analyze their interactions. Strategies combining multiple distinct fluorescent labels are increasingly used to study spatial relationships among cells and molecules and to encode parameters such as neuronal connectivity1,2,3,4,5,6,7,8,9, cell lineage10,11,12,13,14,15, cycling state16,17, subtype identity18, genotype19,20, or signaling pathway activation21. Scaling up such approaches at the whole organ or tissue level would be a major step forward but is hindered by the lack of suitable large-volume multicolor microscopy methods. In recent years, serial block-face imaging, which relies on the automated, iterative alternation of imaging and microtome-based sectioning of a block of tissue, has been successfully transposed from electron microscopy to light microscopy22,23,24,25. This scheme has emerged as an effective means for generating data encompassing mm3-to-cm3 volumes of tissue at subcellular-resolution with either discrete or continuous sampling. One particularly active field of application is neuroscience, where block-face fluorescence imaging has opened the way to brain-wide mesoscale connectomics23,26 and single neuron reconstruction efforts26,27,28,29. However, microtome-assisted microscopy methods developed so far are limited to single- or dual-color imaging23,24,26,29. This limitation is due to the general difficulty of exciting a manifold of fluorescent proteins. In addition, achieving tissue-scale multicolor microscopic imaging requires addressing chromatic aberrations and channel registration over large volumes. These obstacles made it so far very difficult to probe cell interactions or more generally to image multiplexed or combinatorial fluorescent signals with micrometer-scale precision in samples exceeding a few hundreds of microns in depth.
Here, we report on a method for volume multicolor and multi-contrast microscopy with submicrometer registration of the image channels. Our approach, termed chromatic multiphoton serial (ChroMS) microscopy, relies on the integration of trichromatic two-photon excitation by wavelength mixing (WM)30 with automated serial tissue sectioning. We show that ChroMS microscopy delivers multicolor imaging over >mm3 volumes with constant micron-scale resolution and submicron channel coregistration, which sets new quality standards for large-scale multicolor microscopy. We demonstrate its performance for tridimensional imaging of mouse brains labeled by transgenic, electroporation-based or viral multicolor approaches. In addition to being perfectly suited for applications based on combinatorial fluorescence labeling, ChroMS microscopy also enables organ-wide imaging of label-free nonlinear signals such as third harmonic generation (THG) and coherent anti-Stokes Raman scattering (CARS). We illustrate its potential for high information-content three-dimensional (3D) imaging by demonstrating (i) analysis of astroglial cell morphology and contacts over several mm3 of cerebral cortex, (ii) color-assisted tracing of tens of pyramidal neurons in a densely labeled, mm-thick cortical sample, and (iii) brain-wide color-based multiplexed mapping of axonal projection trajectories and interdigitation.
Together, these results show that astrocytes present significant heterogeneity in the volume of their domains and in their local partitioning of the neural tissue, suggesting that they develop in a plastic rather than fixed manner and may adapt to both their local environment and neighbors during cortical development and maturation. Our findings illustrate the unique strengths of continuous multicolor 3D imaging with ChroMS microscopy for unbiased quantitative 3D analysis of cell morphology and contacts within large tissue volumes. Specific methodological advantages of ChroMS for that purpose include the following: (i) the color contrast increases the throughput of individual cell analysis for a given sample by alleviating the need to use very sparse labeling; (ii) it enables one to analyze contacts between cells labeled with different colors; (iii) the serial multiphoton approach suppresses the orientation and size artefacts due to section registration or tissue shrinking/swelling encountered respectively in histology and clearing techniques. 2b1af7f3a8