PMC

Inference pipelines. (a) Sequential processing of a video input. (b) Pipeline of sequential processing. Each process is separated with a queue. (c) Batch inference pipeline. Input data are stacked on a queue, and inference fetches a batch of images. Lined boxes refer processes that make use of GPU while dotted boxes refer those of CPU.

Service queue. Job requests submitted by the user or initiated by the system maintenance procedures are queued for processing. Execution priorities are determined by user-initiated load, process type, and, in some cases, user privilege. The cloud control server communicates the availability of tasks to available compute nodes on the back end, which then retrieve the relevant data from the user data store and run executable computations locally. Parallel jobs and sequential processes communicate data and messages through inter-process communication data pool.

(A) FIB currents are aligned and calibrated, and the sample is loaded into the FIB-SEM instrument. (B) A target cell is identified on the grid with the SEM and FIB. (C) To correct for errors in mechanical stage movements, backlash correction of the stage is performed. The resulting stage location is saved in the stage navigator. (D) The stage is randomly moved out of position by the user. Using the saved coordinates in the stage navigator, the stage is autonomously moved back to the target. (E) The accuracy of this autonomous stage movement is determined by the user. If the target is not centered in the FIB image, backlash correction is repeated until accurate targeting is achieved (C–E). (F/F’) Rough milling, polishing and drift correction patterns are placed onto the image. Rough milling and polishing patterns are saved separately to the queue. The procedure (B–F’) is repeated to select additional targets. (G/H) Rough milling and lamellae polishing are executed automatically. (I) The grids with milled lamellae are removed and stored.

Part I. Direct observation of ATPase complex queued and moving along dsDNA. Cy3 conjugated gp16 was incubated with (A, B, E) and without (D) dsDNA, tethered between two polylysine beads where (C, F) are magnified images of the framed regions of (B, E), respectively. (A–C) are overlapped pseudocolor images indicating the binding of Cy3-labeled gp16 along the To-Pro-3 stained dsDNA chain (Red: Cy3-gp16; Green: To-Pro-3 DNA). (G, H) The motion of the Cy3-gp16 spot was analyzed and a kymograph was produced to characterize the ATPase walking. (Actual motion videos can be found in the and at http://nanobio.uky.edu/movie.html). Part II. Negatively stained transmission electron microscopy images of ATPase queued along dsDNA. gp16 was bound to non-specific dsDNA in queue. Part III. Recording of two Cy3-gp16/dsDNA complexes showing motionless gp16 spots in a buffer containing no ATP. (A) Sequential images of the recording. (B) Kymograph of the two spots.