2–3 month old animals were anesthetized by intraperitoneally injection of ketamine (100 mg/kg) and xylazine (10 mg/kg). We also administered buprenorphine HCl (0.1 mg/kg) intraperitoneally as a pre-emptive analgesic. Once subject mice were in deep anesthesia, they were immobilized in a Kopf stereo-taxic apparatus using intra-aural positioning studs and a tooth bar to immobilize the skull. Heat is provided for warmth by a standard heating pad. We drilled a hole on the surface of the skull at 1.94 mm anterior to Bregma and 0.39 mm lateral for injection into the prefrontal cortex. Using a 33 G Nanofil needle and World Precision Instrument Nanofil syringe at a depth of [–2.95 mm, we injected 1 ul (1 × 1013 viral genome copies) AAV into the right hemisphere of the brain. Injection rates were monitored by the World Precision Instruments UltraMicroPump3. After injection, we closed the incision site with 6-0 Ethilon sutures (Ethicon by Johnson & Johnson). Animals were postoperatively hydrated with 1 ml lactated Ringer's solution (subcutaneous) and housed in a temperature controlled (37°C) environment until achieving ambulatory recovery. Meloxicam (1–2 mg/kg) was also administered subcutaneously directly after surgery. For downstream analysis, EGFP+ tissue was dissected under a stereotactic microscope.
Ultramicropump 3
Manufactured by World Precision Instruments
Sourced in United States
The UltraMicroPump III is a precision syringe pump designed for accurate and controlled fluid delivery. It features a stepper-motor-driven syringe for precise volume control and flow rate management. The device is capable of dispensing volumes ranging from nanoliters to milliliters.
Automatically generated - may contain errors
Lab products found in correlation
Nanofil syringe, by World Precision Instruments (6 mentions)
Micro4 controller, by World Precision Instruments (6 mentions)
Stereotaxic frame, by Kopf Instruments (2 mentions)
Sch39166, by Bio-Techne (2 mentions)
Rhodamine b base, by Merck Group (2 mentions)
4 0 vicryl suture, by Johnson & Johnson (1 mentions)
Cyclopentolate, by Akorn (1 mentions)
Wt 129s1 svlmj, by Jackson ImmunoResearch (1 mentions)
C57bl 6j mice, by Jackson ImmunoResearch (1 mentions)
Micro4 pump controller, by World Precision Instruments (1 mentions)
Paav u6 gfp vector, by Cell Biolabs (1 mentions)
Rompun, by Bayer (1 mentions)
Stereotaxic frame, by Stoelting (1 mentions)
Aav helper free packaging system, by Cell Biolabs (1 mentions)
Micro4 micro syringe pump controller, by World Precision Instruments (1 mentions)
Isoflurane, by Fujifilm (1 mentions)
Gentamicin, by Merck Group (1 mentions)
Stereotaxic frame, by Narishige (1 mentions)
24 protocols using ultramicropump 3
1
Viral Vector Injection in Mouse PFC
2
Poloxamer Delivery via Round Window Injection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice were anesthetized with an intraperitoneal injection of ketamine (100 mg/kg) and xylazine (20 mg/kg) and positioned right ear down. A 12-mm postauricular skin incision was made, and subcutaneous tissues and superficial fascia of the neck were bluntly dissected. After exposing the otic bulla, tympanotomy was performed using a 0.6 mm diamond burr, and the hole was enlarged using microforceps until the round window membrane was clearly visible. A pulled glass micropipette (P0674-1PAK, Sigma-Aldrich) was positioned within the round window niche, and 1 μl of the poloxamer solution was injected using UltraMicroPump 3 (World Precision Instruments). The tympanotomy hole was sealed with muscle, and the wound was closed with 4-0 vicryl sutures (Ethicon).
3
Generating Rd8-free Abca4-/- Rdh8-/- Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Abca4−/−Rdh8−/− mouse strain was a kind gift from Dr Krzysztof Palczewski (26 (link)). To eliminate Rd8 mutation, Abca4−/−Rdh8−/− mice were crossbred with WT 129S1/SvlmJ (Jackson Laboratory) to generate Abca4−/−Rdh8−/− mice without Rd8 mutation in the 129S1/SvlmJ background. Age-matched WT 129S1/SvlmJ mice were used as control for experiments involved in Abca4−/−Rdh8−/− mice. C57BL/6J mice were purchased from Jackson Laboratory to verify ADAM17 as the VLDLR sheddase in vivo. All procedures using experimental animals strictly followed the protocols approved by the Institute Animal Care and Use Committee at the University of Oklahoma Health Sciences Center. All experiments were performed to comply with the Use of Animals of Association for Research in Vision and Ophthalmology guidelines. Mice were anesthetized with an intraperitoneal injection of a ketamine–xylazine mix (ketamine, 100 mg/kg; xylazine, 5 mg/kg). The pupils were dilated with topical application of cyclopentolate (Akorn). For intravitreal injection, the treatment was delivered using a 33-gauge needle attached to a Nanofil syringe (World Precision Instruments) through an incision on the sclera posterior to the limbus. Ultra–micropump III (World Precision Instruments) was applied to control injection speed at 0.5 μl/s. After injection, the antibiotic ointment was applied to the sclera to prevent possible infection.
4
Optogenetic Targeting of Interneurons in Mouse Cortex
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Plasmid pAAV-EF1α-DIO-hChR2(H134R)-P2A-EYFP was generated by replacing the hChR2(C128A H134R) in pAAV-EF1α-DIO-hChR2(C128A H134R)-P2A-EYFP (Prakash et al., 2012 (link)) with the hChR2(H134R) from pAAV-EF1α-DIO-hChR2(H134R)-EYFP (Addgene #20298) and was deposited at Addgene (#139283). The recombinant AAV vectors were produced by the Gene Vector Core at Baylor College of Medicine. To express ChR2 in Pv or Sst interneurons, 200 nl of AAV9-EF1α-DIO-hChR2(H134R)-P2A-EYFP vectors (3 × 1013 genome copies/ml) were injected into the somatosensory cortex of Stxbp1tm1d/+;PvCre/+ and Stxbp1+/+;PvCre/+ or Stxbp1tm1d/+;SstCre/+ and Stxbp1+/+;SstCre/+ mice, respectively, at postnatal day 1–5 as previously described (Xue et al., 2014 (link); Messier et al., 2018 (link)) with an UltraMicroPump III and a Micro4 controller (World Precision Instruments).
5
Striatal Injection: Minimally Invasive Technique
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice were anesthetized with isoflurane and positioned on a stereotaxic frame (David-Kopf Instruments, Tujunga, CA). A 1-cm midline incision was made in the skin over the dorsal surface of the skull, and the skull was exposed to allow the positioning of a drill over the bregma point of reference. From the bregma, the coordinates used were +1 mm anteroposterior, +1.7 mm lateral, and −3 mm ventral to the skull surface. These coordinates were selected to target the center of the left striatum and avoid passing through any ventricle. BHs were injected with Nanofil syringes (World Precision Instruments, Sarasota, FL) and steel bevel needles (33-gauge diameter; World Precision Instruments) into the striatum at a rate of 0.25 µl/min with a total of 0.5 µl per mouse controlled with a pump (UltraMicroPump III with a Micro4 pump controller; World Precision Instruments). The needle was kept in place for 2 min following injection to avoid any reflux of the BH solution. The skin incision was closed with sutures. These conditions produced minimal mechanical trauma to the brain. The patency of the needles was verified prior to and after injections. Mice were euthanized by isoflurane anesthesia overdose, followed by cervical dislocation, at different times after injection.
6
AAV-Mediated Gene Silencing in Mouse Habenula
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The AAV-sh-vehicle and AAV-sh-CAPS2 were constructed using pAAV-U6-GFP vector (provided by Cell Biolabs, Inc., San Diego, CA, USA), containing a U6 promoter expressing shRNA and a PGK promoter expressing EGFP. The sequences of CAPS2 shRNA were 5′-GCTCCATTACAGCTTTGCATT-3′ (Supplementary Table S1 , U6-sh-CAPS2-1), 5′-CCCAGATTCATCTCGAAAGAA-3′ (U6-sh-CAPS2-2), and 5′-GCGCTGCAAATGTTCGTCTTT-3′ (U6-sh-CAPS2-3). Detailed methods for the cell culture and in vitro knockdown experiments are in supplementary material. For viral injection, eight weeks-old mice were anesthetized with 1% isoflurane and placed in a stereotaxic apparatus (Ultra-precise stereotaxic instruments for mice; Stoelting Co., Wood Dale, IL, USA). Mice were injected bilaterally with 1 μl of 1012 to 1013/ ul concentrated AAV viral solution into the MHb (coordinates from bregma: − 1.34 mm anterior/posterior, ± 0.83 mm medial/lateral, − 3.05 mm dorsal/ventral, with 10° angle toward the midline in the coronal plane) using microinjection cannula (30 gauge, Plastics One, Roanoke, VA, USA) and UltraMicroPump III (World Precision Instruments, Sarasota, FL, USA) at 100–120 nl/min. Behavior experiments were performed at least 14 days after surgery. The injection sites were examined at the end of the behavior tests, and only data from animals with correct injections were included.
7
Cortical Ischemia Induction Protocol
8
Local Perfusion of Pharmacological Reagents
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Local perfusion of pharmacological reagents was performed according to a previous study (Zheng et al., 1994 (link)), with some modifications. The drugs were injected into the culture medium by a microinjector (UltraMicroPump III; World Precision Instruments, Inc.) with a flow rate of 3–5 nl/min, which produced lower pressure and caused less disturbance to the PAA gel than the Picospritzer (Parker Inc.) used previously. The inner diameter of the micro pipette tip is ∼1–2 µm.
9
Intravitreal Injection of Mab2F1
10
Intravitreal Transplantation of Cell Suspensions in Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice were first anesthetized by 5% isoflurane (Isoflurin; Axience) inhalation for 2 min and were maintained in deep anesthesia with 2% isoflurane inhalation. Oxybuprocaine was also administered locally before transplantation. Cell suspension (1 μl) containing 200,000 cells in MACS buffer was delivered by intravitreal injection. Injection was performed using a micropump (UltraMicroPump III with Micro4 Controller; World Precision Instruments). Cells were delivered at 150 nl/s using a 10-μl NanoFil syringe with a 33G beveled tip (NanoFil). From 1 week before transplantation to the end of the follow-up, mice were maintained under immunosuppression through cyclosporine treatment (210 mg/l) in the drinking water. A total of 27 female mice were analyzed 1 week after transplantation (n = 17 animals with grafted THY1-positive cells; n = 4 animals with grafted unsorted cells) and 4 weeks after transplantation (n = 6 animals with grafted THY1-positive cells).
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!