설치류에 살아서 Stereotaxic 외과

If I undertand the study correctly, it would be interesting to report the results of rate of reuptake for neurotransmitter, which is the main purpose of the empriment. 

Calculations of the reuptake rate of neurotransmitter can indeed be accomplished through net-flux microdialysis. However, the primary objective is the measurement of basal extracellular levels of neurotransmitters and their metabolites.

The authors have wonderfully demonstrated how to perform the stereotaxic experiment in rats. However they should have added few more words on how the ear bars should be adjusted so that it shows equal readings on both the sides before the opening of skull. In my experience I have noted that before opening the skull, one should make sure that both the vernier scales of the ear bars show almost correct readings in order to make sure the skull is on the right path, failure of which might lead to the miscalculation of the stereotaxic coordinates.

Thanks and Regards,

Rajesh S Omtri.

Correct placement of the ear bars is clearly a practice effect. We usually have one of the ear bars tight in position, then insert the tight ear bar in the ipsilateral ear canal, hold it in place and slowly insert the loose ear bar on the contralateral side before tightening it down. It is desirable that the skull is centered in between the ear bars. The skull surface must be always level (parallel to the platform of the stereotaxic instrument and at 90° to the guide of the microdialysis cannula) and skin at the incision surface should be flat and present no humps. These problems occur if the ear bars are inserted incorrectly (not in the ear canal). Correct ear bar placement can be identified by gently trying to move the head of the animal up and down and left to right. Before tightening the incisor bar, up or down movement but not lateral movement should be possible. Correct placement of the ear bars in the ear canal is the most important prerequisite for accurate stereotaxic placement.

Emmanuel Pothos 

I am a scientist and I find it very hard to see such a gruesome procedure like this one. There should be a clear label on the content of videos as they can be quite disturbing, and they shouldn't be automatically broadcasted on the main website page. I wonder if the editors of this journal seriouly consider the possibility of risks the authors might face by being attacked by animal activists, and if the Principal Investigators of similar papers are held liable for exposing their students' identity to those groups while making these videos.

This message does not intend to diminish the value of the present work, but to bring this serious issue to the attention of the editors and the authors who appear on the video.

All procedures described in this article have been reviewed by the Institutional Animal Care University Committee at Tufts Medical Center and approved as compliant with federal and state standards of animal care. JoVE also conducted a veterinary review of the article before publication; nothing was "automatically broadcasted" as the viewer claims. Animals are anesthetized before any type of brain surgery, carefully monitored for appropriate depth of anesthesia and hydration during the procedure and diligently followed up through postoperative care with analgesic medication and antibiotics until full recovery. Stereotaxic brain surgery is one of the most sophisticated procedures in live and survivable animal surgery and it normally involves minimal pain for the operated animal because of the conditions set in place as described above. Stereotaxic brain electrode placement is a procedure that has been routinely used even for humans (i.e. Parkinson's disease patients) and such operations have been repeatedly broadcasted over the Internet from several hospitals for educational purposes. In some of these cases, the discomfort of the patient is so minimal that general anesthesia is not used and the patient is awake during surgery and able to respond to questions from the surgeons, who use the patient's response to assess the accurate placement of the electrode in the brain. The whole process in animals or humans is elegant, effective and high technology driven, not gruesome and painful.

We appreciate the concern of the viewer about safety issues, but scientists have to take responsibility for their own work and it is not appropriate to publish it anonymously, being in this journal or elsewhere. Otherwise, the whole concept of the validity of peer-reviewed research and accountability of authors for their work is negated. There are numerous pieces of published work in different journals, including dissection videos, autopsies of animal tissue, images of animals etc. that can potentially be used by extremists to target the authors. Censoring scientific journals and scientists cannot eliminate this possibility.

Stereotaxic surgery should be performed under aseptic conditions. The surgeon shoud have a cap, mask, and surgery gloves. She should not be touching non sterile items while doing surgery, i.e. pens, cannula etc. Ophthalmic ointment is essential. Hemostats are not good  skin retractors as they damage tissue. There are antibiotics that can be given subcutaneously, which is easier and less painful to give.

There is not such a thing as sterile stereotaxic surgery in living animals. The mere presence of a living animal on the table with its fur, bodily fluids etc. negates sterile conditions. Doing the procedure under a culture hood with negative air flow is also not advisable as it limits access to the animal from all angles, it makes it more difficult for the animal to maintain appropriate temperature due to the air flow and it contaminates the hood area, which is counterintuitive particularly if the hood is used for cell cultures. The most appropriate actions are to sterilize the components used for the surgery (i.e. cannula and skull screws) prior to use, sterilize all insrtruments before surgery and during surgery as needed and maintain as clean of an environment as possible in the incision area by shaving away the fur and treating with povidine prior to the incision. Gloves should be used, face mask and cap will not hurt but none of the above will ensure sterile conditions.

There is a variety of skin retractors available, we have not found that hemostats are worse than others in damaging tissue.

Antibiotics given subcutaneoulsy are acceptable, but not as long lasting as those given intramuscularly. In any case, the easiest antibiotic to administer is bacitracin, right around the headcup of the animal.

Emmanuel

Suggestion for dental cement:  My lab uses a UV dental acrylic that is much easier to handle.  The acrylic sets when exposed to a UV light in about 10 seconds, and we do not need to use bone screws to secure the cap.  However, I'm guessing that the UV acrylic is more expensive.  Its available from Pentron.

Oh, and don't forget eye lube.

We have tried in the past to use dental cements that their manufacturer claims do not require head screws.  We were not convinced. In many cases the cement head cup came off as one piece as we were trying to implant the microdialysis probe. Using sterile head screws is the best way to ensure that the cement cup will be securely attached to the skull. Any other method shaves off about 20 min from each surgery but it increases the probability that the cup will come off and waste the entire procedure. Suppliers do tend to charge more for cements that supposedly work without head screws so in the long run this is not cost effective. 

Eye lube as an eye protectant is indeed a very good precaution for this procedure. 

Nice demonstration of stereotaxic surgery in rats.

I think that the best way to control that the skull is perfectly flat (parallel to the platform) would to check the height coordinates at the bregma and at the lambda using the canula as recommended in the stereotaxic atlas. That might not be a problem for ICV canulation since the ventricle are quite big but for canulation in a specific structure or nucleus it is critical. 

I usually use only 4 screws but I guess 6 are necessary for a microdialysis probe.

Also, do you calculate the coordinates from the surface of the skull or from the dura ?

We calculate steotactic coordinates from the skull surface.

how can i download (Survivable Stereotaxic Surgery in Rodents) thanks

Hi.  Please contact us at support@jove.com.

ı can not understand that why u r doing such as these trials for understanding brain mechanism, cuz I believe that if somethings can not explained naturally, we also can not understand exactly

Hi

              I am khalid a Ph.D scholar in deptt of pharmacy,university of Peshawar Pakistan.Its really great contribution to science and i eally enjoyed and learnt alot from the movie of Survivable Stereotaxic Surgery in Rodents

thanks indeed and keep up this great work.

khalid rauf

Wow, you guys really do not knwo what you are doing.

Why would you use the archaic acrylic dental cement when you could use Glass Ionomer Luting Cement?

Why didnt you anesthiatize with O2 delivered isofluorine?

Why did you not sue a stereotaxic drill?

Why was the cement applied so sloppy?

Why do you not use a digittal display for the coordinates, it ensures much more precise surgeries.

Hi Dave,

How does the Glass Ionomer Luting Cement compare with the Light-cured Dental Adhesive Resin listed in this journal by Okamura lab?

Look forward to hearing from  you.

Thanks,

Jim

In our hands, dental acrylic is the only cement that ensured headcups stayed on for several weeks when used in combination with 6 skull screws. Emmanuel