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.
Nvista 2
Manufactured by Inscopix
Sourced in United States
NVista 2.0 is a miniaturized, head-mounted microscope designed for in vivo calcium imaging of neural activity in freely behaving animals. It allows researchers to capture high-resolution images and videos of neuronal activity in real-time.
Automatically generated - may contain errors
Lab products found in correlation
Nvista 3, by Inscopix (2 mentions)
Nvoke, by Inscopix (1 mentions)
Aav5 cag dlight1, by Addgene (1 mentions)
Leptin, by Merck Group (1 mentions)
Phosphate buffered saline (pbs), by Merck Group (1 mentions)
Ethovision xt version 12, by Noldus (1 mentions)
I o box, by Noldus (1 mentions)
Nvista hd, by Inscopix (1 mentions)
Nvoke2, by Inscopix (1 mentions)
Data acquisition software, by Inscopix (1 mentions)
12 protocols using nvista 2
1
In Vivo Calcium Imaging of Mouse MGB
2
Miniature Microendoscope Baseplate Attachment
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At least 2 wk after GRIN-lens implantation, the baseplate of a miniature microendoscope (nVista 2.0, Inscopix) was attached over the GRIN lens by a conventional method (27 (link)). Briefly, the mice were anesthetized and mounted onto the stereotaxic device described above, and a baseplate attached to the miniature microscope was placed on the GRIN lens by using Gripper (Inscopix). The optimal location was determined by monitoring the fluorescent images of GCaMP-expressing neurons (where the largest number of neurons were in focus), and the baseplate was fixed with dental resin cement (Super-Bond C&B, Sun Medical) at this position. In cases where we failed to identify neurons at this stage (usually due to the failure of GRIN-lens implantation), the baseplate was not attached, and such mice were excluded from the experiment. After baseplate attachment, a baseplate cover was placed on the baseplate until Ca2+ imaging was performed.
3
Miniaturized Microscopy Imaging of Neural Activity
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For miniaturized microscopy imaging, D1R-cre, D2R-cre and A2a-cre transgenic mice were injected with an AAV encoding the calcium indicator GCaMP6f (AAV5-hSyn-FLEX-GCaMP6f, University of Pennsylvania, Vector Core) into the Nucleus Accumbens ( + 1.7 mm AP, ± 0.7 mm ML, –4.4 mm DV from bregma), or dorsal striatum ( + 1.4 mm, ± 1.5 mm, –2.4 mm, respectively). For fiber photometry recordings, wildtype mice were injected into the NAc with an AAV encoding the dopamine sensor dLight (AAV5-CAG-dLight1.1, Addgene). Afterwards a gradient refractive index (GRIN) lens (0.5 mm or 0.6 mm diameter for NAc, 1.0 mm diameter for dStr, Inscopix) or an optic fiber (0.4 mm diameter, MFC_400/430–0.48_4 mm_ZF2.5(G)_FLT, Doric Lenses) was placed above the viral injection site and secured to the skull using dental cement. After 4–6 weeks, a baseplate that holds the miniscope (nVista 2.0 or nVista 3.0, Inscopix) was attached to the implant.
4
In Vivo Ca2+ Imaging After Leptin
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Ca2+ imaging was performed in the homecage. After attachment of the microendoscope (nVista2.0, Inscopix, Palo Alto, CA), animals were given 5 min to acclimatize to the microendoscope. Ca2+ imaging was performed for 5 min, 25 min after i.p. injection of PBS (Sigma/Merck, Darmstadt, Germany) or leptin (5 mg/kg; Sigma/Merck, Darmstadt, Germany).
5
In Vivo Calcium Imaging of DG Neurons
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Ca2+ imaging of DG neurons was performed while the mice freely traveled in an OF or T-maze, each on a different day. Each different group of mice was used for an OF or T-maze. Prior to Ca2+ imaging, the mice were lightly anesthetized, and a miniature microscope (nVista 2.0, Inscopix) was mounted onto the baseplate of the mice. The mice were then placed back in the home cages and then transferred to a sound-proof behavioral experiment room. At least 30 min after recovery from anesthesia, the mice were subjected to OF or T-maze tests, while the Ca2+ signals of their DG neurons were obtained at a 3-Hz sampling rate with 1,440- × 1,080-pixel resolution. We applied 475/10-nm LED light for the excitation of GCaMP6f fluorescence (∼0.24 mW/mm2 at the bottom of the GRIN lens).
6
Cranial Window Implantation and GCaMP6f Delivery in Mice
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Adult male and female mice (WT (n = 3; female = 1) and cKO (n = 3; female = 1); 14–18-weeks old) were anesthetized using isoflurane and placed in a stereotaxic frame for AAV2/5 GfaABC1D-GCaMP6f delivery and cranial window implantation as previously described [3 (link)]. A 3 mm craniotomy was made over the frontal cortex leaving the dura intact. AAV2/5 GfaABC1D-GCaMP6f (3.31 × 1013 GC/mL; Penn Vector Core, Philadelphia, PA, USA) was injected at two adjacent sites (1.5 µL each site; 200 nl/min) in the frontal cortex (AP: 2.0–2.5 mm, ML: −1.25–−1.75 mm, DV: −0.18 mm). The needle remained in place for 10 min after each injection. After vector delivery, a 3 mm glass coverslip was fixed over the craniotomy with cyanoacrylate adhesive and the skull covered with dental acrylic. After 2–4 weeks recovery, mice were fitted with a baseplate under isoflurane anesthesia. This recovery window allowed enough time to detect the fluorescent indicator [3 (link),63 (link),64 (link)]. The baseplate was secured to the skull with dental acrylic mixed with black carbon powder to house the miniature microscope (nVista 2.0; Inscopix, Palo Alto, CA, USA).
7
Neural Activity Imaging in Mouse mPFC
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We use a head-mounted miniaturized microscope (nVista 2.0 and nVoke 1.0, Inscopix, Palo Alto, CA) to record the GCaMP6m fluorescent signals from mPFC neurons. The increase of GCaMP6m signal reflects a burst of spikes/action potentials54 ; the number of spikes has been experimentally determined for certain cell types26 (link), though not, as yet, for mPFC neurons. The microscope was mounted onto the mouse’s head right before imaging and was triggered by a TTL pulse from the Topscan system to simultaneously acquire fluorescent signal and behavioral video. The imaging data were acquired at a frame rate of 15 Hz and at 1024 × 1024 pixels. The LED power was set to 0.3–1 mW and the gain was 1 to 2 depending on fluorescence intensity.
8
In vivo Calcium Imaging in Freely Behaving Mice
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In vivo calcium imaging in freely behaving mice was performed using the nVista 2.0 miniature endoscope (Inscopix) following the procedures described in detail elsewhere (Resendez et al., 2016 (link)). Young adult male and female GRov and WT mice underwent injection of AAV5-CamKIIa-GCamp6f (Addgene; 300 nl diluted 1:5 in artificial CSF AP −2.05, ML 1.75 from bregma; DZ −1.3 from skull) and implantation of a 4 × 1 mm GRIN lens (at AP −1.95, ML 1.6 from bregma, DZ −1.55 from skull) over the dorsal hippocampal CA1 pyramidal cell layer (Fig. 1A,B ).
For recordings, mice were allowed to explore a large (72 × 72 cm) brightly lit (200 lux) open field for 10 min. Behavior was recorded and analyzed using Noldus Ethovision XT (version 12) and synchronized with calcium imaging using TTL pulses with a Noldus I/O Box. At least 3 days after the completion of behavioral testing, mice were perfused with PBS followed by 4% paraformaldehyde and brains were postfixed in paraformaldehyde for 24 h before removal of the lens. 40-μm sections were cut on a cryostat and used to verify GCamp6f expression in dorsal CA1 pyramidal cells and accurate placement of the lens over the dorsal CA1 region.
For recordings, mice were allowed to explore a large (72 × 72 cm) brightly lit (200 lux) open field for 10 min. Behavior was recorded and analyzed using Noldus Ethovision XT (version 12) and synchronized with calcium imaging using TTL pulses with a Noldus I/O Box. At least 3 days after the completion of behavioral testing, mice were perfused with PBS followed by 4% paraformaldehyde and brains were postfixed in paraformaldehyde for 24 h before removal of the lens. 40-μm sections were cut on a cryostat and used to verify GCamp6f expression in dorsal CA1 pyramidal cells and accurate placement of the lens over the dorsal CA1 region.
9
Miniaturized Imaging and Fiber Photometry in Transgenic Mice
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For miniaturized microscopy imaging D1R-cre, D2R-cre and A2a-cre transgenic mice were injected with an AAV encoding the calcium indicator GCaMP6f (AAV2.5-hSyn-FLEX-GCaMP6f, University of Pennsylvania Vector Core) into the Nucleus Accumbens (+1.7 mm AP, +-0.7 mm ML, -4.4 mm DV from bregma), or dorsal striatum (+1.4 mm, +-1.5 mm, 2,4 mm, respectively).
For fiber photometry recordings wildtype mice were injected into the NAc with an AAV encoding the dopamine sensor dLight (AAV2.5-CAG-dLight1.1, Addgene). Afterwards a gradient refractive index (GRIN) lens (0.5mm or 0.6mm diameter for NAc, 1.0 mm diameter for dstr, Inscopix) or an optic fiber (0.4 mm diameter, MFC_400/430-0.48_4 mm_ZF2.5(G)_FLT, Doric Lenses) was placed above the viral injection site and secured to the skull using dental cement.
After 4-6 weeks a baseplate that holds the miniscope (nVista 2.0 or nVista 3.0, Inscopix) was attached to the implant.
For fiber photometry recordings wildtype mice were injected into the NAc with an AAV encoding the dopamine sensor dLight (AAV2.5-CAG-dLight1.1, Addgene). Afterwards a gradient refractive index (GRIN) lens (0.5mm or 0.6mm diameter for NAc, 1.0 mm diameter for dstr, Inscopix) or an optic fiber (0.4 mm diameter, MFC_400/430-0.48_4 mm_ZF2.5(G)_FLT, Doric Lenses) was placed above the viral injection site and secured to the skull using dental cement.
After 4-6 weeks a baseplate that holds the miniscope (nVista 2.0 or nVista 3.0, Inscopix) was attached to the implant.
10
Micro-Endoscopic Imaging of Social Behavior
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