Advanced Fluorescence Microscopy Techniques-FRAP, FLIP, FLAP, FRET and FLIM

Fluorescence microscopy provides an efficient and unique approach to study fixed and living cells because of its versatility, specificity, and high sensitivity. Fluorescence microscopes can both detect the fluorescence emitted from labeled molecules in biological samples as images or photometric data from which intensities and emission spectra can be deduced. By exploiting the characteristics of fluorescence, various techniques have been developed that enable the visualization and analysis of complex dynamic events in cells, organelles, and sub-organelle components within the biological specimen. The techniques described here are fluorescence recovery after photobleaching (FRAP), the related fluorescence loss in photobleaching (FLIP), fluorescence localization after photobleaching (FLAP), Forster or fluorescence resonance energy transfer (FRET) and the different ways how to measure FRET, such as acceptor bleaching, sensitized emission, polarization anisotropy, and fluorescence lifetime imaging microscopy (FLIM). First, a brief introduction into the mechanisms underlying fluorescence as a physical phenomenon and fluorescence, confocal, and multiphoton microscopy is given. Subsequently, these advanced microscopy techniques are introduced in more detail, with a description of how these techniques are performed, what needs to be considered, and what practical advantages they can bring to cell biological research

Change blindness: eradication of gestalt strategies

Arrays of eight, texture-defined rectangles were used as stimuli in a one-shot change blindness (CB) task where there was a 50% chance that one rectangle would change orientation between two successive presentations separated by an interval. CB was eliminated by cueing the target rectangle in the first stimulus, reduced by cueing in the interval and unaffected by cueing in the second presentation. This supports the idea that a representation was formed that persisted through the interval before being 'overwritten' by the second presentation (Landman et al, 2003 Vision Research 43149–164]. Another possibility is that participants used some kind of grouping or Gestalt strategy. To test this we changed the spatial position of the rectangles in the second presentation by shifting them along imaginary spokes (by ±1 degree) emanating from the central fixation point. There was no significant difference seen in performance between this and the standard task [F(1,4)=2.565, p=0.185]. This may suggest two things: (i) Gestalt grouping is not used as a strategy in these tasks, and (ii) it gives further weight to the argument that objects may be stored and retrieved from a pre-attentional store during this task

Eye-tracking the emergence of attentional anchors in a mathematics learning tablet activity

Little is known about micro-processes by which sensorimotor interaction gives rise to conceptual development. Per embodiment theory, these micro-processes are mediated by dynamical attentional structures. Accordingly this study investigated eye-gaze behaviors during engagement in solving tablet-based bimanual manipulation tasks designed to foster proportional reasoning. Seventy-six elementary- and vocational-school students (9-15 yo) participated in individual task-based clinical interviews. Data gathered included action-logging, eye-tracking, and videography. Analyses revealed the emergence of stable eye-path gaze patterns contemporaneous with first enactments of effective manipulation and prior to verbal articulations of manipulation strategies. Characteristic gaze patterns included consistent or recurring attention to screen locations that bore non-salient stimuli or no stimuli at all yet bore invariant geometric relations to dynamical salient features. Arguably, this research validates empirically hypothetical constructs from constructivism, particularly reflective abstraction

Aerospace Medicine and Biology: A continuing bibliography with indexes, supplement 140

This bibliography lists 306 reports, articles, and other documents introduced into the NASA scientific and technical information system in March 1975

Micro Fourier Transform Profilometry (μ\muFTP): 3D shape measurement at 10,000 frames per second

Recent advances in imaging sensors and digital light projection technology have facilitated a rapid progress in 3D optical sensing, enabling 3D surfaces of complex-shaped objects to be captured with improved resolution and accuracy. However, due to the large number of projection patterns required for phase recovery and disambiguation, the maximum fame rates of current 3D shape measurement techniques are still limited to the range of hundreds of frames per second (fps). Here, we demonstrate a new 3D dynamic imaging technique, Micro Fourier Transform Profilometry ($\mu$FTP), which can capture 3D surfaces of transient events at up to 10,000 fps based on our newly developed high-speed fringe projection system. Compared with existing techniques, $\mu$FTP has the prominent advantage of recovering an accurate, unambiguous, and dense 3D point cloud with only two projected patterns. Furthermore, the phase information is encoded within a single high-frequency fringe image, thereby allowing motion-artifact-free reconstruction of transient events with temporal resolution of 50 microseconds. To show $\mu$FTP's broad utility, we use it to reconstruct 3D videos of 4 transient scenes: vibrating cantilevers, rotating fan blades, bullet fired from a toy gun, and balloon's explosion triggered by a flying dart, which were previously difficult or even unable to be captured with conventional approaches.Comment: This manuscript was originally submitted on 30th January 1

Medical SLAM in an autonomous robotic system

One of the main challenges for computer-assisted surgery (CAS) is to determine the intra-operative morphology and motion of soft-tissues. This information is prerequisite to the registration of multi-modal patient-specific data for enhancing the surgeon’s navigation capabilities by observing beyond exposed tissue surfaces and for providing intelligent control of robotic-assisted instruments. In minimally invasive surgery (MIS), optical techniques are an increasingly attractive approach for in vivo 3D reconstruction of the soft-tissue surface geometry. This thesis addresses the ambitious goal of achieving surgical autonomy, through the study of the anatomical environment by Initially studying the technology present and what is needed to analyze the scene: vision sensors. A novel endoscope for autonomous surgical task execution is presented in the first part of this thesis. Which combines a standard stereo camera with a depth sensor. This solution introduces several key advantages, such as the possibility of reconstructing the 3D at a greater distance than traditional endoscopes. Then the problem of hand-eye calibration is tackled, which unites the vision system and the robot in a single reference system. Increasing the accuracy in the surgical work plan. In the second part of the thesis the problem of the 3D reconstruction and the algorithms currently in use were addressed. In MIS, simultaneous localization and mapping (SLAM) can be used to localize the pose of the endoscopic camera and build ta 3D model of the tissue surface. Another key element for MIS is to have real-time knowledge of the pose of surgical tools with respect to the surgical camera and underlying anatomy. Starting from the ORB-SLAM algorithm we have modified the architecture to make it usable in an anatomical environment by adding the registration of the pre-operative information of the intervention to the map obtained from the SLAM. Once it has been proven that the slam algorithm is usable in an anatomical environment, it has been improved by adding semantic segmentation to be able to distinguish dynamic features from static ones. All the results in this thesis are validated on training setups, which mimics some of the challenges of real surgery and on setups that simulate the human body within Autonomous Robotic Surgery (ARS) and Smart Autonomous Robotic Assistant Surgeon (SARAS) projects

Medical SLAM in an autonomous robotic system

One of the main challenges for computer-assisted surgery (CAS) is to determine the intra-operative morphology and motion of soft-tissues. This information is prerequisite to the registration of multi-modal patient-specific data for enhancing the surgeon’s navigation capabilities by observing beyond exposed tissue surfaces and for providing intelligent control of robotic-assisted instruments. In minimally invasive surgery (MIS), optical techniques are an increasingly attractive approach for in vivo 3D reconstruction of the soft-tissue surface geometry. This thesis addresses the ambitious goal of achieving surgical autonomy, through the study of the anatomical environment by Initially studying the technology present and what is needed to analyze the scene: vision sensors. A novel endoscope for autonomous surgical task execution is presented in the first part of this thesis. Which combines a standard stereo camera with a depth sensor. This solution introduces several key advantages, such as the possibility of reconstructing the 3D at a greater distance than traditional endoscopes. Then the problem of hand-eye calibration is tackled, which unites the vision system and the robot in a single reference system. Increasing the accuracy in the surgical work plan. In the second part of the thesis the problem of the 3D reconstruction and the algorithms currently in use were addressed. In MIS, simultaneous localization and mapping (SLAM) can be used to localize the pose of the endoscopic camera and build ta 3D model of the tissue surface. Another key element for MIS is to have real-time knowledge of the pose of surgical tools with respect to the surgical camera and underlying anatomy. Starting from the ORB-SLAM algorithm we have modified the architecture to make it usable in an anatomical environment by adding the registration of the pre-operative information of the intervention to the map obtained from the SLAM. Once it has been proven that the slam algorithm is usable in an anatomical environment, it has been improved by adding semantic segmentation to be able to distinguish dynamic features from static ones. All the results in this thesis are validated on training setups, which mimics some of the challenges of real surgery and on setups that simulate the human body within Autonomous Robotic Surgery (ARS) and Smart Autonomous Robotic Assistant Surgeon (SARAS) projects

Exploiting Temporal Image Information in Minimally Invasive Surgery

Minimally invasive procedures rely on medical imaging instead of the surgeons direct vision. While preoperative images can be used for surgical planning and navigation, once the surgeon arrives at the target site real-time intraoperative imaging is needed. However, acquiring and interpreting these images can be challenging and much of the rich temporal information present in these images is not visible. The goal of this thesis is to improve image guidance for minimally invasive surgery in two main areas. First, by showing how high-quality ultrasound video can be obtained by integrating an ultrasound transducer directly into delivery devices for beating heart valve surgery. Secondly, by extracting hidden temporal information through video processing methods to help the surgeon localize important anatomical structures. Prototypes of delivery tools, with integrated ultrasound imaging, were developed for both transcatheter aortic valve implantation and mitral valve repair. These tools provided an on-site view that shows the tool-tissue interactions during valve repair. Additionally, augmented reality environments were used to add more anatomical context that aids in navigation and in interpreting the on-site video. Other procedures can be improved by extracting hidden temporal information from the intraoperative video. In ultrasound guided epidural injections, dural pulsation provides a cue in finding a clear trajectory to the epidural space. By processing the video using extended Kalman filtering, subtle pulsations were automatically detected and visualized in real-time. A statistical framework for analyzing periodicity was developed based on dynamic linear modelling. In addition to detecting dural pulsation in lumbar spine ultrasound, this approach was used to image tissue perfusion in natural video and generate ventilation maps from free-breathing magnetic resonance imaging. A second statistical method, based on spectral analysis of pixel intensity values, allowed blood flow to be detected directly from high-frequency B-mode ultrasound video. Finally, pulsatile cues in endoscopic video were enhanced through Eulerian video magnification to help localize critical vasculature. This approach shows particular promise in identifying the basilar artery in endoscopic third ventriculostomy and the prostatic artery in nerve-sparing prostatectomy. A real-time implementation was developed which processed full-resolution stereoscopic video on the da Vinci Surgical System

Aerospace Medicine and Biology: A continuing bibliography with indexes, supplement 165, March 1977

This bibliography lists 198 reports, articles, and other documents introduced into the NASA scientific and technical information system in February 1977