Chapter 26 – Multiple Color Single Molecule TIRF Imaging and Tracking of MAPs and Motors

Microtubules are part of a complex mechano-chemical network inside cells. In order to understand how the components of these systems work together, careful in vitro experiments must be performed with added complexity. These experiments can ideally image all the interacting species. In order to image these molecules, multiple-color fluorescence imaging can be performed. In this chapter, we describe some methods for performing multiple-color single molecule fluorescence imaging using total internal reflection fluorescence microscopy. We give several specific examples of species of microtubule-associate proteins and motors that can be examined with detailed protocols for labeling, purification, and imaging

Chapter 27 – Studying Plus-End Tracking at Single Molecule Resolution Using TIRF Microscopy

The highly dynamic microtubule plus-ends are key sites of regulation that impact the organization and function of the microtubule cytoskeleton. Much of this regulation is performed by the microtubule plus-end tracking (+TIP) family of proteins. +TIPs are a structurally diverse group of proteins that bind to and track with growing microtubule plus-ends in cells. +TIPs regulate microtubule dynamics as well as mediate interactions between microtubule tips and other cellular structures. Most +TIPs can directly bind to microtubules in vitro; however, the mechanisms for their plus-end specificity are not fully understood. Cellular studies of +TIP activity are complicated by the fact that members of the +TIP family of proteins interact with each other to form higher-order protein assemblies. Development of an in vitro system, using minimal components, to study +TIP activity is therefore critical to unequivocally understand the behavior of individual +TIP proteins. Coupled with single molecule imaging, this system provides a powerful tool to study the molecular properties that are important for +TIP function. In this chapter, we describe a detailed protocol for in vitro reconstitution of +TIP activity at single molecule resolution using total internal reflection fluorescence microscopy

The experience of couples being given an oxygen concentrator to use at home: A longitudinal interpretative phenomenological analysis

This longitudinal study explores the lived experience of four couples where one person from each couple is prescribed an oxygen concentrator to use at home. Transcripts were analysed using interpretative phenomenological analysis. The findings reported here focus on two super-ordinate themes: ‘the journey of acceptance’ and ‘negotiating changing relationships’. Participants described a gradual process of accepting the device into their lives, the impact on couple’s relationships and the role that expectations have in mediating that process. These themes suggest that patient education that considers the psychological and social issues may prove useful in facilitating the acceptance process

Two New Gecko Species Allied to Bavayia sauvagii and Bavayia cyclura (Reptilia: Squamata: Diplodactylidae) from New Caledonia

Two new species of the diplodactylid gecko Bavayia are described from Mt. Koghis, Province Sud, New Caledonia. One species is a large, characteristically colored representative of the B. sauvagii complex. It is sympatric with B. sauvagii itself, for which a neotype is here designated. The second new taxon is a large member of the Bavayia cyclura group. Selection of a neotype of B. sauvagii and designation of a lectotype of B. cyclura facilitate future evaluation of intra- and interspecific variation within these two species groups. Although restricted in apparent range, both new species are relatively common where they occur

Triggering social interactions:chimpanzees respond to imitation by a humanoid robot and request responses from it

Even the most rudimentary social cues may evoke affiliative responses in humans and promote socialcommunication and cohesion. The present work tested whether such cues of an agent may also promotecommunicative interactions in a nonhuman primate species, by examining interaction-promoting behavioursin chimpanzees. Here, chimpanzees were tested during interactions with an interactive humanoid robot, whichshowed simple bodily movements and sent out calls. The results revealed that chimpanzees exhibited twotypes of interaction-promoting behaviours during relaxed or playful contexts. First, the chimpanzees showedprolonged active interest when they were imitated by the robot. Second, the subjects requested ‘social’responses from the robot, i.e. by showing play invitations and offering toys or other objects. This study thusprovides evidence that even rudimentary cues of a robotic agent may promote social interactions inchimpanzees, like in humans. Such simple and frequent social interactions most likely provided a foundationfor sophisticated forms of affiliative communication to emerge