Överleva Stereotaxic Kirurgi hos gnagare (Translated to Swedish)

Cite this Article: Överleva Stereotaxic Kirurgi hos gnagare

Geiger, B. M., Frank, L. E., Caldera-Siu, A. D., Pothos, E. N. Survivable Stereotaxic Surgery in Rodents . J. Vis. Exp. (20), e880, doi:10.3791/880 (2008).

Abstract: Överleva Stereotaxic Kirurgi hos gnagare

Förmågan att mäta extracellulära basala nivåer av signalsubstanser i hjärnan hos vakna djur gör det möjligt för bestämning av effekterna av olika systemiska utmaningar (farmakologisk eller fysiologiska) till CNS. Till exempel kan man mäta direkt hur djurets mitthjärnan dopamin prognoser svara på dopamin-frisättande läkemedel som d-amfetamin eller naturliga stimuli såsom mat. I denna video visar vi hur du implantat vägleda kanyler inriktas på specifika platser i råtta hjärnan, hur du infogar och implantat en mikrodialys sond och hur du använder höga kromatografi prestanda vätska i kombination med elektrokemisk detektion (HPLC-EG) för att mäta extracellulära nivåer av oxiderbara signalsubstanser och metaboliter. Lokala exakt införandet av droger genom mikrodialys sond möjliggör förfinad arbete på plats specificitet i en sammansatt s verkningsmekanism. Denna teknik har utmärkta anatomiska och kemiska upplösning men bara blygsamma tidsupplösning som mikrodialys prover brukar behandlas varje 20-30 minuter för att se till detekterbara signalsubstansen nivåer. Kompletterande ex vivo-verktyg (dvs, skiva och cellodling elektrofysiologi) kan hjälpa till med övervakning i realtid neurotransmission.

Protocol: Överleva Stereotaxic Kirurgi hos gnagare

Sammanfattning

Två månader gammal medelålder C57BL/6J möss eller motsvarande eller tre månader gammal medelålder Sprague Dawley råttor eller motsvarande bedövas med ketamin (60 mg / kg ip för råtta, 100 mg / kg ip för möss) och xylazin (10 mg / kg, ip för antingen arter). Sedering övervakas med en mjuk tå nypa tillbaka reflex visats i Walantus et al. (Jove, 6, 2007) och Szot et al. (Jove, 9, 2007). Värmereglering kan ske genom en thermostatregulated värmedyna (ALA Instruments Inc.) och övervakas genom en rektal termometer. Huvudet är rakat av päls och rengöras med jod innan snitt. Efter att huden snitt (2 cm lång för råttor, 1 cm långa för möss) och avlägsnande av alla mjukvävnad från ytan av skallen, är placeringen av guiden kanyl bestämmas i förhållande till bregma. Ett 6 mm hål borras genom skallen med en batteridriven borraren avsedd för gnagare kirurgi (Fine Science Tools, Inc.). Man ser till så att borret inte tränga igenom meningeal membran eller blodkärl. Skull implanteras med bilateral 5 mm 21 gauge rostfritt axlar stål guide som leder till den bakre kärnan accumbens, dorsala striatum eller prefrontala cortex. Den stereotaxic koordinater är etablerade enligt Franklin och Paxinos, 1997 (Musen Brain i Stereotaxic Koordinater, Academic Press) eller Paxinos och Watson, 2006 (The Rat Brain i Stereotaxic Koordinater, Academic Press). Implantat är säkrade genom dentala cement. En bolusdos av laktat Ringers av 0,9% koksaltlösning ges vid slutet av operationen (5mls SC hos råttor och 1 ml SC hos möss efter vätskor värms till normal kroppstemperatur) för att förhindra uttorkning. Buprenorfin (0.1-0.5mg/kg SC) ges två gånger dagligen och då, på ett behov när det behövs, om djuret verkar vara i smärta. Lokal behandling med antibiotika (bacitracin salva) och systemisk behandling med antibiotika (penicillin 100.000 IE / kg IM var 12 timmar för de första 48 timmarna efter op) ges om postoperativa infektioner uppstå.

Efter operation, är djur i individuella boxar med mat och vatten tillgängligt ad lib. Minst en vecka är tillåtet för återhämtning innan mikrodialys och eutanasi. Efter återhämtningen från operationen, djuren överförs till ett mikrodialys bur och sonder mikrodialys införas, och cementerade i guiden schakt som har installerats under operation. Probe insättning orsakar inte smärta eller obehag på grund av att givaren är förbi hud, muskler och meningeal vävnad genom guiden axeln. Därför är sond insättning görs utan bedövning och alla anestesi-inducerade effekter på neurokemi eller beteende undviks. Vi låter sonderna stabiliseras i 12 timmar och då vi börjar provtagningen var 30 minuter för ytterligare 8-12 timmar beroende på experimentet. Vi bevakar rörelseapparaten djurets beteende via fotoceller eller manuell inspelning av rörelse av försöksledaren. Microdialysate prover injiceras i en High Performance Liquid Chromatography med elektrokemisk detektion (HPLC-EG) instrument för neurokemiska detektion och analys. Vi ser för effekter på basal neurokemi och motorisk beteende. I slutet av försöket djuret avlivas genom en överdos av systemisk ketamin (200 mg / kg ip) och xylazin (20 mg / kg, ip). Då hjärtat är perfusion med 0,9% koksaltlösning följt av 4% paraformaldehyd. Hjärnan tas bort, fryst och skär längs med mikrodialys sonden tarmkanalen för att verifiera korrekt sond placering.

Förfarande

Sätt upp stereotaxic instrumentet och allt material som behövs. Se till att området och instrument rengörs och steriliseras.
Raka av päls med elektrisk rakapparat. Gå från öronen för att bara i-mellan ögonen, flytta rakkniv i olika riktningar för att effektivt rent område av päls. Applicera povidine / jod till rakade området men skyddar ögonen från det.
Montera djuret på stereotaxic apparaten genom att placera örat barer i hörselgången och strama på plats. Montera först det ena örat bar i hörselgången, och sedan hålla den på plats och skjut in det andra örat baren. Du vet att du är på rätt plats när huvudet inte längre kan flyttas i sidled. Säkra munnen med den främre fästet av stereotaxic och se till att huvudet är i nivå med en linjal. Sätt linjalen i vertikalt läge med avseende på stereotaxic instrumentet plattformen och kontrollera att en 90 ° vinkel mellan härskaren och mitten av djurets hårbotten). Bekräfta detta genom stereotaxic instrumentet om det finns en sådan förmåga.
Gör en främre / bakre snitt i hårbotten med en steril skalpell sträcker sig från lambda att bara emellan ögonen på djuret. Använd steriliserade peanger att nypa bort från huden och hålla snittet öppen. Använda flera sterila bomullspinnar, torka den exponerade skallen ytan.
Sätt guiden kanylen på dess fäste, hitta bregma på skallen, och placera guiden kanylen rakt över denna plats. Skriv ner främre / bakre och laterala koordinater. Från bregma, hitta rätt koordinater som behövs för placering av din sond med hjälp av stereotaxic Atlas. Placera guiden kanylen till rätt koordinater genom att addera eller subtrahera från bregma. Ta din guide kanyl tills det ligger an mot skallen, och sedan spela in denna ventrala samordna. Gör en penna markerar med en steril penna på denna plats på skallen, det är där du kommer att borrning.
Ta bort guiden kanylen och sterilisera din borr. Försiktigt borra ett hål i blyerts märket tills du kommer fram bredden på skallen. Kontrollera med guiden kanylen för att se om det skulle rensa hålet utan att vidröra sidorna. Håll borrning och kontroll tills kanylen kan klara på en rak väg. När hålet är gjort, använd en steril nål för att försiktigt punsch meningerna i syfte att möjliggöra fri införandet av kanylen.
Nästa, med en handborr, göra sex hål för skallen skruvar: två främre till kanylen hålet, två laterala att kanylen hålet och två bakre åt sidorna. Sterilisera sex skruvar och placera dem på skallen tills de är ordentligt förankrade på.
Rengör guiden kanylen med etanol och koksaltlösning, montera, och sänk den långsamt till rätt ventrala koordinaten. Se till att sidorna inte vidrör och att den går i helt lodrät.
Placera ankaret skruv medialt och bakom den bakre skallen skruvar och håller den på plats med pincett. Blanda en tunn sats av flytande dentala cement och täcker guiden kanylen, skruvar, och resten av skallen med en steril spatel. Gör en ny sats, tjockare denna gång, och helt täcka området och kanylen och förankra skruv tillräckligt för att säkra den.
Som cement blir tjockare och innan det stelnar, separera huden från cement-cupen och mögel cementen koppen med spatel för att se till att cement locket är jämn runt och inte irritera huden senare.
Låt dentala cement för att helt torr innan du tar bort djuret från apparaten. Ta bort peanger. Applicera bacitracin hela vägen runt cementen locket.
När djuret är utanför stereotaxic instrumentet, injicera den med 0,25 ml penicillin IM (intramuskulär) följt av 1 ml koksaltlösning SC (subkutant).
Placera djuret i sin egen bur och bevaka den tills den blir medveten innan det återsänds till sitt rum för att återhämta sig.
Övervaka djur tills de återhämta sig från narkos på operationsdagen och daglig post-op, fram till slutet av försöket, för tecken på infektion och utvärdering av smärta / lidande. Detta inkluderar helger och helgdagar. Låg spontan rörelse, ångest vocalization vid hantering, krökt kroppshållning, diarré, svullnad och ansvarsfrihet området för headmount, och brist på mat / dricka är alla tecken på smärta och ångest. Buprenorfin (0.1-0.5mg/kg SC) administreras två gånger dagligen, och sedan, på ett behov när det behövs, om djuret verkar vara i smärta. Lokal behandling med antibiotika (bacitracin salva) och systemisk behandling med antibiotika (penicillin 100.000 IE / kg IM var 12 timmar för de första 48 timmarna efter op) ges om postoperativa infektioner uppstå. Om något av dessa symtom kvarstår efter administrering av buprenorfin, extra vätska och antibiotikabehandling inom 12 timmar efter kirurgi, är djuret avlivas.

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Discussion: Överleva Stereotaxic Kirurgi hos gnagare

In vivo mikrodialys är det bästa sättet för att mäta olika signalsubstanser och metaboliter i olika hjärna platser i ett levande djur. Övervakar dock bara extracellulära nivåer av neurochemicals och det ger inte tidsupplösningen att övervaka exocytos av neurotransmittorer i realtid. Genom en version av den teknik som kallas "net-flux", kan den verkliga signalsubstansen koncentration vid en viss plats beräknas, vilket i sin tur kan ge exakta mätningar av signalsubstansen andelen återupptagshämmare genom plasmamembranet transportörer.

Mikrodialys är perfekt att illustrera skillnader i basala extracellulära signalsubstansen nivåer mellan olika grupper av djur (dvs olika genotyper) och dechiffrera effekter av droger eller andra manipulationer på signalsubstansen release.

Införandet av analyser alternativ till HPLC-EG som kapillär zon elektrofores (CZE) tillsammans med fluorescerande upptäckt har ökat tiden lösa in vivo mikrodialys inom ett par minuter per prov.

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Disclosures: Överleva Stereotaxic Kirurgi hos gnagare

Acknowledgements: Överleva Stereotaxic Kirurgi hos gnagare

Med stöd av DK065872 (ENP), en Smith Family Foundation Award of Excellence inom biomedicinsk forskning (ENP), F31 DA023760.

Materials: Överleva Stereotaxic Kirurgi hos gnagare

Materials are described in the protocol document.

References: Överleva Stereotaxic Kirurgi hos gnagare

1. Bungay, P.M., Newton-Vinson P., Isele W., Garris P.A. & Justice J.B. Microdialysis of dopamine interpreted with quantitative model incorporating probe implantation trauma. J. Neurochem 86, 932-946, (2003).

2. Chen, K.C. Effects of tissue trauma on the characteristics of microdialysis zero-net-flux method sampling neurotransmitters. Journal of Theor. Biology 238, 863-881, (2006).

3. Geiger B.M., Behr G.G., Frank L., Caldera-Siu A.D., Beinfeld M.C., Kokkotou E.G., Pothos E.N. Evidence for defective mesolimbic dopamine exocytosis in obesity-prone rats. FASEB Journal, Aug; 22:2740-6,(2008).

4. Pothos, E.N., Creese, I. & Hoebel, B.G. Restricted eating with weight loss selectively decreases extracellular dopamine in the nucleus accumbens and alters dopamine response to amphetamine, morphine and food intake. The Journal of Neuroscience 15, 6640-6650, (1995).

Ask the Author: Överleva Stereotaxic Kirurgi hos gnagare

21 Comments

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.

Posted by: cody dingOctober 10, 2008, 6:14 PM

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.

1.1

Posted by: Emmanuel PothosOctober 10, 2008, 6:45 PM

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.

Posted by: RajeshOctober 12, 2008, 5:30 PM

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

2.1

Posted by: Emmanuel PothosOctober 12, 2008, 6:16 PM

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.

Posted by: AnonymousOctober 20, 2008, 3:55 AM

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.

3.1

Posted by: Emmanuel PothosOctober 20, 2008, 8:30 AM

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.

3.1.1

Posted by: AnonymousFebruary 2, 2009, 4:18 PM

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

3.1.1.1

Posted by: Emmanuel PothosApril 22, 2009, 5:43 PM

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.

Posted by: AnonymousNovember 1, 2008, 10:56 PM

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.

4.1

Posted by: Emmanuel PothosNovember 1, 2008, 11:12 PM

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 ?

Posted by: AnonymousDecember 2, 2008, 11:30 PM

We calculate steotactic coordinates from the skull surface.

5.1

Posted by: Emmanuel PothosDecember 3, 2008, 12:11 AM

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

Posted by: maryamDecember 18, 2008, 5:18 AM

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

8.1

Posted by: AnonymousApril 17, 2009, 11:09 AM

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

Posted by: Ayla ArslanJanuary 7, 2009, 7:54 AM

Hey, it is obvious that you are not Dr. Ayla Arslan, then who are you? it seems like you are one of her students using her name as a nick as she always recommend JoVE during her Biopsychology lectures. :)))))))

11.1

Posted by: celimusti93October 31, 2009, 6:14 PM

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

Posted by: khalid raufFebruary 26, 2009, 10:56 AM

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.

Posted by: DaveMarch 12, 2009, 9:38 PM

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

13.1

Posted by: JimMarch 15, 2009, 12:35 PM

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

13.1.1

Posted by: Emmanuel PothosApril 22, 2009, 4:04 PM

Do you use any preanesthetic medications?

Posted by: Vijay BenadeSeptember 19, 2009, 12:51 PM

Usually not, if the animal suffers from CRD (chronic respiratory disease) you can pretreat with atropine to facilitate breathing. However, CRD is an indication of substandard conditions in the animal colony (infrequent change of bedding, poor air flow etc.). If you have animals with CRD, consult with your veterinarian to improve your facility and check on your source for laboratory animals, whether commercial or another lab, for facility conditions as well.

14.1

Posted by: AnonymousOctober 18, 2012, 6:29 PM

I am an undergraduate at the University of California, Santa Barbara doing an Honor's thesis project on the rat dorsal Raphe nucleus. In my project, I need to implant a cannula into the dRN, but am concerned about profuse sagittal sinus bleeding if I go through the midline. I noticed in other papers they often go into the DRN at about a 30 degree angle, in order to avoid this issue and also to avoid the cerebral aqueduct. As the angled cannula is a more complicated procedure, for me it would be easiest to place the cannula at the midline, and I'm wondering what's the best way to deal with these issues, such as how bleeding is stopped or slowed down, how it can be avoided, how many animals I can expect to lose, etc. Any advice would be much appreciated!

Posted by: Oren OferOctober 27, 2009, 3:54 PM

The angled approach is the best, but if you encounter sagittal sinus bleeding make sure to put in place large cotton tips from a sterile bag, press gently for a few minutes to slow down blood flow and leave on until blood has clotted. Then very carefully remove cotton tip to avoid breaking the blood clot. Although this bleeding would be fatal in humans, it usually is not fatal in rats. Emmanuel

15.1

Posted by: AnonymousOctober 18, 2012, 6:34 PM

nice job.
just some comments:
the membrane (after cuting the skin to expose the skull) should be carefully and totally removed - this decrease the chances of the acrylic fall off.
if you do a small cut, 1 or 2 screw would be enough.
another important thing, regarding guide cannula is that it should be obstruct after surgery so that no reflux happens and nothing enters for this hole - if this happens you can loose all you surgery. If, when you try to put a needle inside for drug injection (p.ex) and it doesnt enters, you can use a some H2O2 to open it (in case of blood coaguation)

Posted by: AnonymousDecember 11, 2009, 4:52 PM

We use obdurators to seal off guide cannulas post-op. We avoid using only 1-2 screws no matter what the size of the incision, this is clearly inadequate anchoring for the headcup and it will come off in a matter of a few days at best. It really pays off to anchor the headcup with as many screws as you can. Emmanuel

16.2

Posted by: AnonymousOctober 18, 2012, 6:46 PM

Oh! another thing...
would be really good to use local anesthetic with vasoconstrictor before cutting the scalpe.
this will minimize animal nociception and will avoid excessive bleeding.
but not toooooo much, other wise wont be god to animals, and we also see some increase of infeccion

Posted by: AnonymousDecember 11, 2009, 4:57 PM

The authors and our attending veterinarian would like to add the following information to the article, which was not otherwise clearly stated or shown and may be of help to readers and viewers:
1) After anesthesia and prior to surgery, eye lubricant was applied to protect the corneas of the animals.
2) Prior to inserting the ear bars into the rats' ears, lidocaine gel was applied to provide analgesia.
3) All rats did receive an initial dose of buprenorphine following the surgeries and then were given subsequent doses on an as needed basis. This was not clear in the video or text.
4) The dose of penicillin given was 100,000 IU/kg.
All of the above measures were approved in our IACUC protocol for this procedure and our attending veterinarian has reviewed and verified these additional comments.

Posted by: Emmanuel PothosApril 28, 2010, 3:07 PM

is it better that ketamine is better than halothan as a anesthetic agent

Posted by: Ravi S.May 14, 2010, 2:16 AM

my just quest is how the hole in the skull is closed/filled? Did you leave it for natural tissue growth or use gelatin or something else.thanks

Posted by: i hatip-al-khatibMay 14, 2010, 9:48 AM

In our case the guide cannula leaves very little space to add anything else. Some of my colleagues are using bone wax or gelatin for larger openings. Emmanuel

20.1

Posted by: AnonymousOctober 18, 2012, 6:41 PM

No flash please

Posted by: AnonymousAugust 8, 2010, 1:31 PM

Overall a nice video, but there are a few things that should be done to improve aseptic technique. The eyes need to be lubricated prior to shaving to protect them from the hair and from dessication. To reduce infections a surgical drape should be used, along with a surgical mask. Lastly, pointing with a sterile instrument would have been better.

Posted by: M Lucas, DVMNovember 5, 2010, 6:34 PM

It is a very nice presentation, I would like to add a little in it . When the animal id fixed with ear bars and the scale on ear bar and the scale of sterotaxic base should be equidistant
See in video your demo point 02:51.

Varsha

Posted by: Varsha SharmaMay 24, 2011, 3:35 AM

Im a 4th year Psych Honours student doing a project that needs me to implant canulas into the infralimbic. I just did my second practice surgery today, and it was terrible. The cannulas were mislocated, it took me an hour to put in 4 bone screws, and they went through the skull, and the dental cement ran into its eyes, and Im just glad that rat was put down before it woke up because there is no way it would have survived. I've always been sort of clumsy, and I have to do 35 of these, and half my year is gone and I dont have time to come up with another project.
So yeah, Im freaking out right now,

Posted by: Nadia May 30, 2011, 8:58 AM

Hi Nadia,
It seems you do not have adequate training and supervision to perform this procedure. It is essential that a member of your laboratory team with extensive experience in stereotactic surgery, if not the primary investigator directly and the head veterinarian for your institution's animal facility, should go over things with you multiple times and actively do the procedure with you before any further attempts. You have to ensure the animal's welfare, lack of pain and recovery during and after the procedure if you wish to be anywhere close to acceptable standards. In my opinion, brain stereotactic surgery is an advanced procedure that should be used only with the maximum of caution and the best of training for undergraduate projects. If the animal facility or your faculty supervisor do not have the time or the skills to train you properly, then it would be best to choose something else for your thesis project. Best, Emmanuel

24.1

Posted by: Emmanuel PothosMay 30, 2011, 10:46 AM

Hi everyone,

Just a quick question, after attaching the ear bar I have realized that it takes me quite a while after making an incision in the skull to expose the bregma and lambda. Any suggestions?

Posted by: Jin P.November 5, 2012, 6:41 PM

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Date Published	10/06/2008
JoVE Section	General
JoVE Issue	20
Comments	21 Comments
View Count	Loading... (Details…)
DOI	doi:10.3791/880

Automatic Translation

Överleva Stereotaxic Kirurgi hos gnagare

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