Confocal Microscopy
confocal microscopy
Dr. Priyanka Narayan
Investigate and modulate the cellular pathways perturbed by neurodegenerative disease risk factors using human induced pluripotent stem cell (iPSC)-derived neural cell types. A postdoctoral position is available in the laboratory of Dr. Priyanka Narayan at the National Institutes of Health (NIH) in Bethesda, USA. Genome wide association studies have identified genetic factors that increase risk for neurodegenerative diseases like Alzheimer’s disease. A number of these risk factors are shared between multiple neurodegenerative diseases with diverse pathologies and clinical presentations. Our lab works on multiple questions including: (1) How do disease risk factors alter the cellular pathways to increase susceptibility to disease processes? (2) Can we identify genetic and chemical modulators of these cellular perturbations to prevent or reverse the detrimental effects of risk factors? We use a combination of genetics, biochemistry, molecular biology, and human iPSC-derived neuronal and glial cell types to answer these questions. We envision that our findings can accelerate the development of novel therapeutic or preventative strategies for neurodegenerative diseases.
Open SPM: A Modular Framework for Scanning Probe Microscopy
OpenSPM aims to democratize innovation in the field of scanning probe microscopy (SPM), which is currently dominated by a few proprietary, closed systems that limit user-driven development. Our platform includes a high-speed OpenAFM head and base optimized for small cantilevers, an OpenAFM controller, a high-voltage amplifier, and interfaces compatible with several commercial AFM systems such as the Bruker Multimode, Nanosurf DriveAFM, Witec Alpha SNOM, Zeiss FIB-SEM XB550, and Nenovision Litescope. We have created a fully documented and community-driven OpenSPM platform, with training resources and sourcing information, which has already enabled the construction of more than 15 systems outside our lab. The controller is integrated with open-source tools like Gwyddion, HDF5, and Pycroscopy. We have also engaged external companies, two of which are integrating our controller into their products or interfaces. We see growing interest in applying parts of the OpenSPM platform to related techniques such as correlated microscopy, nanoindentation, and scanning electron/confocal microscopy. To support this, we are developing more generic and modular software, alongside a structured development workflow. A key feature of the OpenSPM system is its Python-based API, which makes the platform fully scriptable and ideal for AI and machine learning applications. This enables, for instance, automatic control and optimization of PID parameters, setpoints, and experiment workflows. With a growing contributor base and industry involvement, OpenSPM is well positioned to become a global, open platform for next-generation SPM innovation.
“Open Raman Microscopy (ORM): A modular Raman spectroscopy setup with an open-source controller”
Raman spectroscopy is a powerful technique for identifying chemical species by probing their vibrational energy levels, offering exceptional specificity with a relatively simple setup involving a laser source, spectrometer, and microscope/probe. However, the high cost of Raman systems lacking modularity often limits exploratory research hindering broader adoption. To address the need for an affordable, modular microscopy platform for multimodal imaging, we present a customizable confocal Raman spectroscopy setup alongside an open-source acquisition software, ORM (Open Raman Microscopy) Controller, developed in Python. This solution bridges the gap between expensive commercial systems and complex, custom-built setups used by specialist research groups. In this presentation, we will cover the components of the setup, the design rationale, assembly methods, limitations, and its modular potential for expanding functionality. Additionally, we will demonstrate ORM’s capabilities for instrument control, 2D and 3D Raman mapping, region-of-interest selection, and its adaptability to various instrument configurations. We will conclude by showcasing practical applications of this setup across different research fields.
Gap Junction Coupling between Photoreceptors
Simply put, the goal of my research is to describe the neuronal circuitry of the retina. The organization of the mammalian retina is certainly complex but it is not chaotic. Although there are many cell types, most adhere to a relatively constant morphology and they are distributed in non-random mosaics. Furthermore, each cell type ramifies at a characteristic depth in the retina and makes a stereotyped set of synaptic connections. In other words, these neurons form a series of local circuits across the retina. The next step is to identify the simplest and commonest of these repeating neural circuits. They are the building blocks of retinal function. If we think of it in this way, the retina is a fabulous model for the rest of the CNS. We are interested in identifying specific circuits and cell types that support the different functions of the retina. For example, there appear to be specific pathways for rod and cone mediated vision. Rods are used under low light conditions and rod circuitry is specialized for high sensitivity when photons are scarce (when you’re out camping, starlight). The hallmark of the rod-mediated system is monochromatic vision. In contrast, the cone circuits are specialized for high acuity and color vision under relatively bright or daylight conditions. Individual neurons may be filled with fluorescent dyes under visual control. This is achieved by impaling the cell with a glass microelectrode using a 3D micromanipulator. We are also interested in the diffusion of dye through coupled neuronal networks in the retina. The dye filled cells are also combined with antibody labeling to reveal neuronal connections and circuits. This triple-labeled material may be viewed and reconstructed in 3 dimensions by multi-channel confocal microscopy. We have our own confocal microscope facility in the department and timeslots are available to students in my lab.
Shaping colloidal bananas to reveal biaxial, splay-bend nematic, and smectic phases
Colloidal dispersions of rod-like particles are widely accepted as convenient model systems to study the phase behavior of liquid-crystal forming systems, commonly found in LCDs. This is due to the fact that colloidal rods exhibit analogous phase behavior to that of elongated molecules, while they can be directly observed by optical microscopy. Recently, there has been a surge of interest in the liquid crystalline behaviour of so-called bent-core, or banana-shaped, molecules. This is due to their ability to form exotic biaxial nematic phases such as the twist-bend and splay-bend nematic phase, which may be of particular interest inherent to their fast switching response in LCDs. Here, we develop model “banana-shaped” colloidal particles with tunable dimensions and curvature, whose structure and dynamics are accessible at the particle level. By heating initially straight rods made of SU-8 photoresist, we induce a controllable shape deformation that causes the rods to buckle into banana-shaped particles. We elucidate the phase behavior of differently curved colloidal bananas using confocal microscopy. Although highly curved bananas only form isotropic phases, less curved bananas exhibit very rich phase behavior, including biaxial nematic phases, polar and antipolar smectic-like phases, and even the long-predicted, elusive splay-bend nematic phase.