Angle two

Manufactured by Leica

Sourced in Israel, Germany, United States

The Angle Two is a precision angle measuring instrument designed for laboratory and industrial applications. It utilizes advanced digital technology to provide accurate and reliable angle measurements. The core function of the Angle Two is to precisely measure and display angular values.

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Lab products found in correlation

Isoflurane, by Merck Group (1 mentions) Cunningham mouse adaptor, by Stoelting (1 mentions) Tissue adhesive, by 3M (1 mentions) Ethilon nylon suture, by Johnson & Johnson (1 mentions)

15 protocols using angle two

Stereotaxic Surgery for Brain Implantation

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For experiments 2 and 3, approximately 2 weeks after arrival, mice were anesthetized with a ketamine (100 mg/kg) and xylazine (10 mg/kg) cocktail and underwent stereotaxic surgery using the Leica Angle Two (Leica Biosystems, Buffalo Grove, IL). Bilateral 26-gauge cannulae (Plastic One, Roanoake, VA) were implanted into the LH (in reference to bregma, AP: −1.10mmML: ± 1.10mm, and DV: − 5.10mm), the bed nucleus of the stria terminalis (BNST; AP: + 0.25mm; ML: ± 1.10mm; DV: − 4.10mm), or the dorsal striatum (DS; AP: + 0.30mm, ML: ± 1.70mm, DV: − 4.10mm). Mice were allowed to recover for a minimum of one week before the initiation of experimental procedures. Cannulae placements were verified histologically at the end of the experiments, and only animals with bilateral hits were included in analyses. Site-directed infusions were delivered using a 1.0 μl Hamilton micro-syringe controlled by a Harvard Apparatus PHD 2000 micro-infusion pump. Post-infusion, the injector was left in place for at least 1 min to allow for drug diffusion away from the injector.

Sprow G.M., Rinker J.A., Lowery-Gionta E., Sparrow A., Navarro M, & Thiele T.E. (2015). Lateral Hypothalamic Melanocortin Receptor Signaling Modulates Binge-Like Ethanol Drinking in C57BL/6J Mice. Addiction biology, 21(4), 835-846.

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Electrode Implantation for Seizure Recording

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Mice that developed SE after pilocarpine injection and naïve mice without pilocarpine injection were surgically implanted with EEG electrodes for further EEG recording using a Leica Angle Two digital stereotaxic instrument (Leica Biosystems) fitted with a Cunningham Mouse Adaptor (Stoelting, Inc, Downers Grove, IL) one week after pilocarpine injection. Mice were anesthetized in an induction chamber supplied with 4–5% isoflurane (Sigma) mixed with 0.8–1.0 L/min oxygen using a calibrated vaporizer. Animals were then administered continuous isoflurane (1–2%) mixed with oxygen (0.8–1.0 L/min) via snout mask for the duration of the anesthesia. Body temperature was maintained using air-activated iron oxide heat packets. Sterile stainless steel bone screw recording electrodes (diameter 0.5 mm, length 1.1 mm; Plastics One) soldered with a lead wire were placed epidurally through rostral burr holes in the skull (AP 1.75 mm, L +2.3 mm) and reference electrodes were implanted caudal to lambda. Electrodes were cemented in place with a rapid-curing dental cement (DenMat Holdings, Lompoc, CA).

Leung A., Ahn S., Savvidis G., Kim Y., Iskandar D., Luna M.J., Kim K.S., Cunningham M, & Chung S. (2014). Optimization of pilocarpine-mediated seizure induction in immunodeficient Nod-Scid mice. Epilepsy research, 0, 114-118.

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Optogenetic Mapping of Mouse Forepaw M1

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We injected a lentivirus encoding ChR2-eYFP under the synapsin promoter (pLenti-Synapsin-hChR2(H134R)-EYFP-WPRE, generated by Karl Deisseroth) into forepaw S1 identified by intrinsic imaging in P30 male wild-type mice under anesthesia using a computerized stereotactic frame (Angle Two, Leica Microsystems). After 2 weeks, mice were sacrificed and the brain was sliced for histological processing. YFP visibility was enhanced using antibody staining (primary antibody: goat-anti-GFP (ACRIS R1091P); secondary antibody Alexa Fluor 647 anti-goat antibody, Life Technologies), and the axonal projection pattern was examined (Figure S2A). We identified a medial agranular region that correlated spatially well with previous electrical (Hira et al., 2013 (link), Li and Waters, 1991 (link), Tennant et al., 2011 (link)) and optogenetic (Ayling et al., 2009 (link), Harrison et al., 2012 (link)) mapping of forepaw M1 in mice. The M1 recording site was chosen as 1 mm medial and 0.3 mm rostral to the functionally identified forepaw S1 (Figure S2B). Overall, the M1 recording locations were approximately centered on stereotactic coordinates ∼0.5 mm anterior and ∼1.7 mm lateral from bregma.

Estebanez L., Hoffmann D., Voigt B.C, & Poulet J.F. (2017). Parvalbumin-Expressing GABAergic Neurons in Primary Motor Cortex Signal Reaching. Cell Reports, 20(2), 308-318.

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Anesthetized Mice Brain Surgery

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Mice were anesthetized with 2% isoflurane in O₂ by inhalation and mounted into a stereotaxic frame (Angle two, Leica Microsystems) linked to a digital mouse brain atlas. Mice were maintained on 1.5–2% isoflurane during the surgery. Mice that had been injected with AAVs were allowed 1 month to recover and for the viral transgenes to adequately express before undergoing behavioral experiments.

Zhang Z., Ferretti V., Güntan İ., Moro A., Steinberg E.A., Ye Z., Zecharia A.Y., Yu X., Vyssotski A.L., Brickley S.G., Yustos R., Pillidge Z.E., Harding E.C., Wisden W, & Franks N.P. (2015). Neuronal ensembles sufficient for recovery sleep and the sedative actions of α2 adrenergic agonists. Nature neuroscience, 18(4), 553-561.

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Anesthetized Mice Brain Surgery

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Targeted knockdown of VTA MOR in rats

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Rats assigned to control viral groups received infusions of lentivirus that expresses green fluorescent protein (GFP) and a short hairpin RNA (shRNA) that does not target any known rat gene, while rats assigned to VTA MOR knockdown groups received a lentivirus that expresses GFP and a shRNA that targets MOR (shMOR) for RNA interference. Lentiviral constructs were prepared as previously described (Lasek et al., 2007 (link)). The shMOR lentivirus reduces VTA MOR expression by 88–97% (Lasek et al., 2007 (link)). Therefore, the viral titre was diluted by 50% with cold sterile saline to reduce the efficacy. After random assignment to GFP or shMOR knockdown conditions, rats were anesthetized using isoflurane and positioned in a stereotaxic frame (Leica Angle Two; Richmond, IL). The appropriate lentiviral construct (1.0 μl each) was infused bilaterally into the VTA (AP −5.15, ML ±2.15, DV −8.7, Tilt 10°; Paxinos and Watson, 2007 ) at a flow rate of 0.1 μl/min, and allowed to diffuse for 10 min before withdrawal of the syringe (Hamilton; Model 7105 KH; 24 gauge tip; Reno, NV). The accuracy of each infusion was later verified using localization of GFP expression. Subjects were given 7 days to recover before the start of intermittent social stress or handling procedures (Fig. 1A).

Johnston C.E., Herschel D., Lasek A.W., Hammer RP J.r, & Nikulina E.M. (2015). Knockdown of ventral tegmental area mu-opioid receptors in rats prevents effects of social defeat stress: Implications for amphetamine cross-sensitization, social avoidance, weight regulation and expression of brain-derived neurotrophic factor. Neuropharmacology, 89, 325-334.

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Stereotaxic Viral Transduction for Optogenetics

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All surgeries were done using a stereotaxic system (Leica Angle Two). The animals were anesthetized with isoflurane (IsoFlo) for the surgery. The skin was opened before determining striatal injection sites with the stereotaxic system. A small opening was made in the skull above the target location at bregma 0.86 mm, medial-lateral (ML) −1.53 mm (Franklin and Paxinos, 2013 ). Subsequently, the left hemisphere was injected with 300 nl viral solution at dorsal-ventral (DV) −3 mm. After that, the wound was closed and sutured before the animal was left for recovery. The virus rAAV5/EF1a-DIO-hChR2(H134R)-eYFP (UNC Vector Core) was used to express Channelrhodopsin2 (ChR2) for optogenetic stimulation experiments. Serotype 5 provided high neuronal and low transynaptic infection (Zincarelli et al., 2008 (link); Aschauer et al., 2013 (link)). The ChR2(H134R) version was selected because previous studies have shown successful expression and excitation in the same mice strains (Lopez-Huerta et al., 2016 (link)).

Pommer S., Akamine Y., Schiffmann S.N., de Kerchove d'Exaerde A, & Wickens J.R. (2021). The Effect of Serotonin Receptor 5-HT1B on Lateral Inhibition between Spiny Projection Neurons in the Mouse Striatum. The Journal of Neuroscience, 41(37), 7831-7847.

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Transplantation of Human ESC-Derived MGE Cells into Mouse Hippocampus

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Human ESC-derived MGE cells in transplantation medium (HBSS with 10ng/ml GDNF, 10ng/ml BDNF and 20 µM Boc-Asp (OMe) fluoromethyl ketone (BAF)) were injected into hippocampus of mice using a Leica Angle Two digital stereotaxic instrument fitted with a Cunningham Mouse Adaptor. Nod-Scid mice were anesthetized using an induction chamber supplied with 4–5% isoflurane mixed with 0.8-1 L/min oxygen using a calibrated vaporizer. Animals were then administered continuous isoflurane (1–2%) mixed with oxygen (0.8–1.0 L/min) for the duration of anesthesia. Cells were transplanted into the following coordinates: AP 1.75 mm, L ±2.3 mm, V −1.7 mm for the rostral CA3 site; AP 3.25 mm, L ±3.0 mm, V −3.65 mm, −2.9 mm and −2.0 mm for the three caudal sites along the dorso-ventral axis of the hippocampus in this coronal plane. A total of 5 × 10⁴ MGE cells in a 0.5 µl volume were delivered to each target coordinate (0.5 µl/1 min). Animals were removed from the stereotaxic instrument, and an intraperitoneal injection (ip) of buprenorphine (0.03 mg/kg) was administered as an analgesic medication. Topical triple antibiotic ointment was applied to the incision site to prevent infection. During recovery, animals were placed on a heating pad and monitored until recovery of motor function.

Ahn S., Kim T.G., Kim K.S, & Chung S. (2015). Differentiation of human pluripotent stem cells into Medial Ganglionic Eminence vs. Caudal Ganglionic Eminence cells. Methods (San Diego, Calif.), 101, 103-112.

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Viral Delivery in Rat VTA for Stress Research

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After random group assignment, rats were anesthetized using isoflurane and positioned in a stereotaxic frame (Leica Angle Two). The respective viral construct was bilaterally infused (1.0 µl per side) into the VTA (AP −5.0 to −5.3, ML ± 2.1, DV −8.2, tilt 10°; Paxinos & Watson, 2007 ) at a constant flow rate of 0.1 µl/min (Hamilton; Model 7105 KH; 24‐gauge tip). After infusion, the needle remained at the infusion site for 5 min to prevent retrace of the virus. Animals that were assigned to non‐viral control groups received sham surgeries, in which they were anesthetized with isoflurane and were positioned in a stereotaxic frame, but no surgery was performed. Rats recovered in their cages for 2–3 weeks to allow for optimal viral expression before the start of social stress or handling procedures.
Viral infusion site localization was performed using fluorescent immunohistochemistry (described below) after each experiment to ensure accurate viral expression in medial/anterior portions of the VTA, which are known to have a high density of DA neurons (Morales & Margolis, 2017 (link)).

Rudolph M.L., Neve R.L., Hammer RP J.r, & Nikulina E.M. (2020). Enhanced psychostimulant response, but not social avoidance, depends on GluA1 AMPA receptors in VTA dopamine neurons following intermittent social defeat stress in rats. The European Journal of Neuroscience, 55(9-10), 2154-2169.

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Viral Vector Injections and Implantation

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Mice were anesthetized with isoflurane and placed in a stereotaxic apparatus (Leica Angle Two) and the height of bregma and lambda was adjusted to be within a difference of 0.1 mm. AAV vector for SubLDT (0.05-0.1 ml), dDpMe (0.1 ml), lPAG (0.1 ml), and MVe (0.2-0.3 ml) was microinjected at a speed of 0.01-0.03 ml/min using a glass capillary and a syringe pump (KD Scientific 210). Target locations were as follows: SubLDT: (medio-lateral (ML), ± 0.7 mm; antero-posterior (AP), À5.2 mm; dorso-ventral (DV), À4.1 mm) dDpMe: (ML, ± 0.5 mm; AP, À4.5 mm; DV, À3.5 mm), lPAG: (ML, ± 0.5 or +0.5 mm; AP, À4.8 mm; DV, À2.5 mm), MVe: (ML, ± 0.8 mm; AP, À5.9 mm; DV, À4.5 mm). After AAV microinjection, EEG (electroencephalogram) and EMG (electromyogram) electrodes were implanted. Electrodes (stainless steel recording screws) for EEG were implanted epidurally over the parietal area (1 mm anterior to lambda, 1 mm lateral to midline) referenced to the cerebellum area. EMG electrodes were bilaterally implanted into the neck muscle. For ICV injection, cannula implantation was performed as previously described [47] (link). A 27G stainless steel tube (10 mm) was inserted into the fourth ventricle (target: ML, 0 mm; AP, À6.0 mm; DV, À4.0 mm) and EEG/EMG electrodes were implanted as described above.

Kashiwagi M., Kanuka M., Tatsuzawa C., Suzuki H., Morita M., Tanaka K., Kawano T., Shin J.W., Suzuki H., Itohara S., Yanagisawa M, & Hayashi Y. (2020). Widely Distributed Neurotensinergic Neurons in the Brainstem Regulate NREM Sleep in Mice. Current biology : CB, 30(6).

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