The miniature microscope (nVista2.0 or nVoke, Inscopix) was fixed to the base plate on the mouse’s head before the experiment using head-fixation at the head bar on a flying saucer style running wheel. Mice were initially habituated to this procedure. MGB Ca2+ fluorescence was imaged continuously during the behavioural session with the following settings (nVista Software Version: 2.0.4 or nVoke Software Version: 2.1.10): Framerate: 20 Hz, blue LED-Power: 50–70%, Gain: 1.0–2.5, nVoke stimulation LED power: 10 mW/mm2. Image size: 1024 × 1024 or 1080 × 1080 pxl. LED power and gain were adjusted according to GCaMP expression levels and the same settings were used across days for individual mice. For all-optical imaging and optogenetic stimulation experiments (nVoke, Inscopix), the stimulation LED was switched on 2 s before the start of the CS and terminated 2 s after the end of the US.
Nvoke
Manufactured by Inscopix
NVoke is a miniaturized fluorescence microscope designed for in vivo imaging of neural activity in freely behaving animals. It provides high-resolution, real-time monitoring of neuronal dynamics with cellular-level resolution.
Automatically generated - may contain errors
5 protocols using nvoke
1
In Vivo Calcium Imaging of Mouse MGB
2
Bulk and Single-cell Calcium Imaging
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For bulk calcium imaging, we used a Doric Lenses fibre photometry system. In the experiment, 465 nm and 405 nm LED light sources (Doric LED driver) were delivered continuously through a rotary joint (Doric Lenses, FRJ_1×1_PT-400/430/LWMJ-0.57_1m) connected to the patch cord (Doric Lenses, MFP_400/430/1100-0.57_1m), and the GCaMP6 signal was collected back through the same fibre into the photodetector (Doric Lenses). For single-cell calcium imaging, we used nVoke (Inscopix).
3
In Vivo Calcium Imaging in Kiss1 Neurons
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For calcium imaging, Kiss1-Cre mice injected with Cre-dependent GCAMP6s AAVs were implanted with Proview Integrated Lenses that combine a 500-µm diameter, 8.4-mm-length GRIN lens with a baseplate (1050-004611, 0.5-mm diameter, 8.4-mm length; Inscopix Inc.) designed for attachment of a miniaturized, single-photon fluorescent microscope (nVoke; Inscopix Inc.). First, a 600-µm-diameter optical fiber (FT600UMT; Thorlabs Inc.) with a sharpened tip was inserted into polyimide tubing (624-µm diameter, cut to 8.4-mm length, 141-0159; Nordson Medical). The polyimide-optical fiber pairing was attached to the stereotaxic frame and implanted 200 to 300µm above the arcuate nucleus. The implant was angled at 6° toward midline to reduce motion artifacts from proximity to the third ventricle. The polyimide tubing was secured to the skull using C&B metabond dental cement (375-0407; Parkell, Inc.) and, once dry, the optical fiber was retracted. The integrated GRIN lens was inserted through the polyimide tubing until 200 µm above the ARC and the baseplate was secured to the skull using C&B dental cement. After surgery, mice were housed individually and allowed to recover for at least 1 wk before receiving daily habituation for microscope attachment and serial blood sampling. No gross behavioral abnormalities were observed in animals that received surgery.
4
In Vivo Ca2+ Imaging in NAc Neurons
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An integrated microscope (nVoke, Inscopix, Palo Alto, California) was used to image GCaMP6s in NAc neurons through an implanted 7 mm GRIN lens (Part number 1050-002179, Inscopix, Palo Alto, California). Grayscale HDF5 images (12 bit) were acquired using nVoke Acquisition Software (version 2.1, Inscopix, Palo Alto, California) at 15 frames per second with an average exposure time of 66.7 ms. The analog gain on the image sensor was set between 1.5 and 3.0, while the EX-LED was maintained within 0.5–1.2 mW/mm2 (measured from the bottom of the objective). Ca2+ imaging, optogenetic activation, and behavior was time-stamped and synchronized.
5
In Vivo Calcium Imaging Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
1-photon calcium imaging was performed with nVoke and nVista microscopes (Inscopix) at a frame rate of 20 Hz (exposure time: 49.90 ms, gain: 2–5 (mean: 2.6 ± 0.2), LED power: 0.3–1.1 mW/mm2 (mean: 0.7 ± 0.1 mW/mm2)). The imaging plane was set to a depth to allow imaging of the highest number of cells. In each imaging session mice performed ~40 trials over the course of 15 min. Landmarks such as blood vessels and the location of active cells were used to maintain a consistent field of view (FOV) over days. Calcium imaging and behavioral recording were synchronized either by triggering the microscope by behavioral video acquisition (EthoVision XT 11.5) or by triggering a blue LED in the FOV of the behavioral camera for offline alignment with the calcium data.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!