Abstract: Provided herein are methods for minimizing Z-image acquisition comprising receiving a first set of three-dimensional (3D) positional information of a plurality of biological molecules within a sample, wherein the first set of 3D positional information is within a first imaging volume and based on a probing cycle of the sample in an imaging instrument, wherein the probing cycle comprises generating optical signals corresponding to at least some of the plurality of biological molecules; determining, based on the first set of 3D positional information, a second imaging volume that is less than the first imaging volume; and directing the imaging instrument to image the second imaging volume in at least one subsequent probing cycle of the sample.

Abstract: Various embodiments of the present disclosure disclose methods and systems for actively monitoring an opto-fluidic instrument for vibrational disturbances and correcting images obtained during the detected disturbances. In various embodiments, an optical imaging system may acquire Z-stack images of samples supported by an XY-stage. Vibrations can cause individual stacks to be sheared, i.e., to not be co-located with respect to neighboring Z-stacks. In various embodiments, an offset between the measured positions and the expected positions of the sheared stacks may be computed, and a determination as to whether to correct or reacquire the Z-stack images may be made based at least in part on the computed offset.