Journal: Journal of Biomedical Optics

Article Title: Red-shifted excitation enhances the sensitivity of red genetically encoded Ca 2+ indicator and enables crosstalk-free two-photon holographic optophysiology

doi: 10.1117/1.JBO.30.11.116003

Figure Lengend Snippet: Optical setup of 2P holographic microscope. Schematic diagram of the optical setup integrating holographic stimulation with 2P imaging (a), a photograph of a 2P holographic microscope (b), the calculation process for producing a 3D spot using an SLM (c), the MATLAB-based GUI for generating two-photon spots (d), and a representative fluorescent image (e) are shown.

Article Snippet: First, two-dimensional fluorescent images are obtained by a 2P scanning microscope system (C2 plus, Nikon) including control software (NIS Elements, Nikon) and a tunable femtosecond (fs) laser from 660 to 1300 nm at a pulse width of 100 fs and a repetition rate of 80 MHz (Chameleon Discovery NX, Coherent, Saxonburg, PA). shows a representative obtained fluorescent image.

Techniques: Microscopy, Imaging

Journal: Nature Communications

Article Title: Open-source, high performance miniature 2-photon microscopy systems for freely behaving animals

doi: 10.1038/s41467-025-62534-y

Figure Lengend Snippet: a Optomechanical design of the system, including all housings, lenses, optical filters, and on-board PCBs in cross-section. The red beam denotes the 920 nm excitation, and the green beam path describes the fluorescent collection onto the detector head. b Exploded view of the UCLA 2P Miniscope, describing the various sub-assemblies that work together, and the individual parts that comprise them. c Mechanical model of the microscope showing external appearance once assembled. d Assembled microscope under test at UCLA.

Article Snippet: After 14 days, expression levels were assessed using a benchtop 2P microscope (Scientifica VivoScope) fitted with a Nikon 16x/0.8NA water immersion objective.

Techniques: Microscopy

Journal: Nature Communications

Article Title: Open-source, high performance miniature 2-photon microscopy systems for freely behaving animals

doi: 10.1038/s41467-025-62534-y

Figure Lengend Snippet: a Simplified block diagram of the custom and off-the-shelf electronic components. The custom PCBs communicate with one another via thin coaxial cables and the specifics of the connections can be seen. b UCLA 2P Miniscope interface PCB. This single low-cost 2 layer PCB integrates electronic signals from various sources and packages it into a single connector; the microscope plugs into this connector. This interface PCB receives four daughter boards: two SiPM drive modules (Hamamatsu), an I 2 C controller (NI), and the MEMS amplifier (Mirrorcle). The interface PCB receives inputs from ScanImage-compatible electronics and additional hardware. c UCLA 2P Miniscope flex PCB. These electronics are a fundamental part of the headpiece and are used to control the MEMS scanning mirror and electrotunable lens.

Article Snippet: After 14 days, expression levels were assessed using a benchtop 2P microscope (Scientifica VivoScope) fitted with a Nikon 16x/0.8NA water immersion objective.

Techniques: Blocking Assay, Microscopy, Control

Journal: Nature Communications

Article Title: Open-source, high performance miniature 2-photon microscopy systems for freely behaving animals

doi: 10.1038/s41467-025-62534-y

Figure Lengend Snippet: a Schematic drawing of the imaging conditions, including the titanium cranial window implant and the objective lens from the UCLA 2P Miniscope. b 1P image collected in the same animal, in the same brain region on the same day as all other panels except E. 1P Image was collected with a custom-made benchtop 1P microscope with a 0.5 NA and high resolution scientific Complementary Metal Oxide Semiconductor (sCMOS) image sensor. c Experimental mouse in the behavioral chamber, during the >20-min imaging session. Light-blue line shows a subset of the animal trajectory in the chamber over time. d Imaging results from the microscope system over the course of the free behavior experiment. e Extracted footprints of the active neurons within the FOV (randomly-colored) following motion correction, overlayed on the maximum intensity projection image. f Neuropil subtracted activity from the neurons in ( g , h ). g Firing locations of a subset of neurons from ( f , h ) plotted within the behavioral arena. h 25 example statistically significant place cells (single sided, multiple comparison corrected, based on Shannon Information content ).

Article Snippet: After 14 days, expression levels were assessed using a benchtop 2P microscope (Scientifica VivoScope) fitted with a Nikon 16x/0.8NA water immersion objective.

Techniques: Imaging, Microscopy, Activity Assay, Comparison

Journal: Journal of Neural Engineering

Article Title: Metaplasticity and continual learning: mechanisms subserving brain computer interface proficiency

doi: 10.1088/1741-2552/add37b

Figure Lengend Snippet: (A). Schematic illustration of an all optical exogenous BCI experiment. Rewarded neural ensemble activity instantly decoded and used as a control signal drive a ‘neural cursor’ on the screen in the form of orientation gratings. (B). Endogenous BCI experiment uses the instantly decoded fluorescence (e.g. Ca 2+ imaged througha two-photon (2P)microscope at920 nm) to directly photo stimulate the same (or different) neuron(s) co-expressing an opsin in closed loop at a non-overlapping spectral wavelength (1040 nm). (D). Protocol for characterizing BTSP and IP over a full experimental session. The open loop photo stimulation is used to measure the effects of performing the closed loop approach at disjoint intervals. In this open loop step, each cell is photo stimulated with a single depolarizing pulse (4–5 repetitions, separated by 15 s for a total duration of ∼1 min) and the evoked fluorescence Ca 2+ traces are averaged to obtain a baseline for a proxy of the global dendritic plateau potential before the closed loop step (optical clamp). The open loop step is repeated at multiple time points following cessation of the clamp. The parameters \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{upgreek} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} $\alpha $\end{document} α e and τ e are used to characterize the changes in the plateau potential’s amplitude and time constant observed in each case. (C). Endogenous BCI operation using an optical clamp: the decoded fluorescence is compared to a user specified threshold target activity level A CL , either in the same cell or in other cells within the field of view (FOV). If the activity drops below the target level, photo stimulation is delivered to depolarize the cell keep its activity above that level and the cycle continues for a user specified interval T CL .

Article Snippet: After allowing 4–6 weeks for expression, animals were head-fixed under the 2P microscope (Bruker Ultima, WI) and the imaged field of view (FOV, 900 μ m × 900 μ m) was screened for regions of interests (ROIs) between ∼100 and ∼250 μ m below the cortical surface that co-express GCaMP7s and ChRmine Kv2.1 .

Techniques: Activity Assay, Control, Fluorescence, Microscopy, Expressing