ZEISS at Society for Neuroscience

Presented by: Leanne Holt, PhD. Postdoctoral Fellow in the laboratory of Dr. Eric Nestler, Icahn School of Medicine at Mount Sinai

Emerging evidence implicates non-neuronal cells in nearly every neuropsychiatric disorder. With the rise of single-cell sequencing approaches, a plethora of investigations are underway to elucidate the transcriptional and epigenetic regulation of these cells, including astrocytes. We have identified one such mechanism involving transcription factor CREB in astrocytes. Given the well-known astrocyte-microglia cross talk, we investigated how CREB-mediated transcriptional regulation may impact microglia morphology utilizing the ZEISS arivis Image Analysis Software. Iba1 staining reveals indeed that viral-mediated modulation of CREB activity in astrocytes modulates microglia morphology.

Presented by: Olivia Prazeres da Costa, PhD. Senior Product Manager ZEISS Microscopy 

Join us to witness the launch of our newest technology designed to accelerate your neuroscience research. Discover how our latest solution pushes the boundaries of imaging.

Presented by: John Mendenhall
In collaboration with A. Wetzel 2, T. Bartol 3, T. Middendorf 1, R. Salzer 4, V. Thiyagarajan 1, M. Kuwajima 1, J. Carson 5, T. Sejnowski 3, K. M. Harris 1

1. The University of Texas at Austin, Center for Learning and Memory 2. Carnegie Mellon University, Pittsburgh 3. Salk Institute for Biological Studies, La Jolla 4. Carl Zeiss Microscopy, Jena 5. Texas Advanced Computing Center

tSEM is a non-destructive volume-imaging method for automated capture of large-field images (> 50k x 50k pixels) at high lateral resolution (~ 2 nm) from serial ultrasections (~ 45 nm thickness). This method uses transmission detection on a <30 kV field emission SEM platform. However, the axial resolution of tSEM through the sample thickness presents a barrier to sub-cellular volume reconstruction by obscuring critical axial details of oblique membranes, intracellular space, synaptic zones, perisynaptic astroglia, smooth ER, multivesicular bodies, spine apparatus, and local axon trajectories, even within the ultrathin sections.

We are developing an automated tomographic method, tomoSEM, that expands the capabilities of tSEM. In this method, the multiple projections acquired per region of interest are transmitted through easy-to-handle, thick (250 - 300 nm) serial sections. Current computed volumes indicate a five-fold improvement in axial resolution over ultrathin section projections that is consistent from the center to the margin across the field diameter. The tomoSEM modifications to a standard SEM are minor and relatively inexpensive. These include automation acquisition software controlling a 360o rotating super-stage with increased distance to a small diameter solid state STEM detector.

This presentation will address crucial aspects of SEM that make tomography a viable volume imaging method such as accurate dynamic focusing across very large and highly tilted fields and the use of very short (<300ns) dwell times that harness the power of averaging across multiple projections. The capability of generalized tilt and rotation combinations not limited to particular axes allows us to explore generalized projection geometries including Fibonacci-style “irrational” acquisition schemes to minimize the number of required images and reduce reconstruction artifacts.

NSF Technology Hub– 1707356

NSF NeuroNex – 2014862

NSF-NCS - 221989

Presented by: Zidan Yang, Graduate Student Laboratory of Dr. Hidehiko Inagaki, Max Planck Florida Institute for Neuroscience

Flexible control of motor timing is crucial for behavior. Before volitional movement begins, the frontal cortex and striatum exhibit ramping spiking activity, with variable ramp slopes anticipating movement onsets. This activity in the cortico-basal ganglia loop may function as an adjustable ‘timer’, triggering actions at the desired timing. However, because the frontal cortex and striatum share similar ramping dynamics and are both necessary for timing behaviors, distinguishing their individual roles in this timer function remains challenging. To address this, we conducted perturbation experiments combined with multi-regional electrophysiology in mice performing a flexible lick-timing task. Following transient silencing of the frontal cortex, cortical and striatal activity swiftly returned to pre-silencing levels and resumed ramping, leading to a shift in lick timing close to the silencing duration. Conversely, briefly inhibiting the striatum caused a gradual decrease in ramping activity in both regions, with ramping resuming from post-inhibition levels, shifting lick timing beyond the inhibition duration. Thus, inhibiting the frontal cortex and striatum effectively paused and rewound the timer, respectively. These findings suggest the striatum is a part of the network that temporally integrates input from the frontal cortex and generates ramping activity that regulates motor timing.