Veteran İşleri Bölümü&#39;nden bir Liyakat Grant ve Sağlık Ulusal Enstitüleri enstitünün bir T32-HL098062 hibe bu çalışmaları destekledi. Biz minnetle Randolph H. Hasting, MD, Ph.D. tavsiyesi teşekkür etmek istiyoruz ve tavsiye ve VA San Diego Sağlık Sistemi Veteriner Tıp Birimi desteği.

1.. Hayvan Preparasyon <ol><li> (Küçük fareler bir uzman tarafından entübe edilebilir) 8 hafta daha eski ve daha 20 g olan fareler edinin. </li><li> Anestezi <ol><li> Periton boşluğu içine bir preanesthetic olarak ketamin ve ksilazin 20 mg / kg, her biri, farelere enjekte edilir. (Bu doz tamamen fare uyutmak için yeterli değildir, ancak mekanik ventilasyon entübasyon sonrası güvenli taşınmasını kolaylaştırır.) Ancak, dozlar ayarlama kurumsal veterinere danışılarak anestezik yanıta göre gerekli olabilir.) </li><li> 90-120 saniye boyunca bir indüksiyon odası içinde% 3.5 izofluran / oksijen ile farelerde anestezi neden. </li><li> Dikkatle, solunum hızı dikkate alınmalıdır. Değil en az 45 bpm giderek yavaş olmalıdır. </li></ol></li></ol> 2. Entübasyon için hazırlık <ol><li> Indüksiyon odasından fareyi çıkarın ve sıkıca Scruff. </li><li> Gent yastıklı ince forseps kullanınly ağız dil uzatmak. </li><li> Iki parmak arasında dil tutma ve yumuşak bir kuvvet uygulanarak dil uzatma koruyun. </li><li> Dikey hareket ile otoskop ve spekulum üzerinde fare yukarı çekin. </li><li> Sıkıca Eksfoliyant ise otoskop oküleri bakmak ve dil ve berduş tarafından yavaşça fareyi yukarı çekin. </li><li> Ses telleri için dikkatle bakmak. Onlar bu zaman kolayca görünür olmalıdır. Laryngis ADITUS (laringeal açılış yapısı) nispeten beyaz bakmak gerekir. Her nefes ile kablolarından hareket olmamalıdır. Kordonlar hareket etmiyor veya kötü görüntülendi varsa, biraz hayvan döndürün ve yavaşça boynunu hyperextend. </li><li> Dominant elin ile, bir kalem gibi, kateterin ucuna uzanan PE10 boru 1 cm uzunluğunda, kateter 20 G, 1 tutun ve spekulumun tarafına takın. PE boru, bir stile veya buji olarak hizmet vermektedir. </li><li> PE10 Tubi yönlendirinng ses tellerinin (stile) ve göbek kadar PE boru üzerinde 20 G kateter (endotrakeal tüp) ilerletmek alt kesici dişlerinin düzeydedir. Hızla bu stile kaldırmak. </li><li> Yavaşça spekulum kapalı hayvan almak ve deney boyunca genel anestezi korumak için sürekli% 2 izofluran ile mekanik ventilatöre hayvan koyarak boru konumunu doğrulayın. Entübasyon onaylamak için PEEP tuzak geçiyor süresi dolan havayı (baloncuklar) gözünüzde canlandırın. Özofagus entübasyon birkaç baloncuklar neden olsa da, o kadar belirgin ya da endotrakeal entübasyon ile tutarlı olmayacaktır. Ayrıca, negatif deplasmanlar için bir hava yolu basıncı izleme (Şekil 1) gözlemlemek. Bunlar, borunun doğru yerleştirilmesi teyit etmektedir. Seçenek olarak ise, IV boru halinde su küçük bir miktar ve ET borusu bağlanabilir. Solunum 2 fare ve gelen hareket doğrulanıyor. Özofagus entübasyon, p durumundaressure izleme anlamlı olarak daha yüksek basınçlar ve hiçbir olumsuz deplasmanlar ortaya çıkaracaktır. Endtidal CO 2 yararlı olmasına karşın, teknik olarak bu küçük tidal hacimleri (~ 200 ul) ve bu çok pahalı olacaktır yapmak için ekipman verilen çok zor olacaktır. Tarif basit teknikleri tam yeterli ve çok daha ucuzdur. Izofluran anestezi mevcut değilse, tüm işlemler, genel anestezi xylazine, 10-20 mg / kg ketamin ile bağlantılı olarak 80-120 mg / kg kullanılarak yapılabilir. </li></ol> 3. Alternatif Tekniği: Doğrudan Larengoskopi Otoskop Tekniği ile Temelde Akciğer içine Çalışma Maddelerin Doğrudan Uygulamasının için, Diğer Amaçlı kolaylıkla Kullanılır. <ol><li> Indüksiyon odası içinde 90-120 saniye boyunca% 3.5, bu işlem için tek başına izofluran ile anestezi hayvanlarda neden. </li><li> Sıkıca kafatası tabanında hayvan Scruff ve yavaşça dil uzatmakince forseps ile. </li><li> Hayvan daha da yukarı çekti olamaz kadar yavaşça dilini tutan, dik spekulum üzerine (değiştirilmemiş) hayvan çekin. </li><li> Ses telleri tek başına bu teknikle en sık görüntülenmiştir, fakat hayvan ve tam görünüme getirmek için hiper-genişletilmiş boyun döndürmek olabilir. </li><li> Glottis instillate içeren bir jel yükleme pipet ilerlemek ve sıvıyı aşılamak. Spekulum hipofarenks ile bir mühür yapar çünkü fare kabloları aracılığıyla gitmez herhangi bir kalıntı sıvı aspire edecek. Bu kolayca rhonchorous tarafından doğrulanmadı nefes zaman sıvı akciğerlerde tam olarak dağıtılmış kadar fare şimdi yapar geliyor. </li><li> Alternatif olarak, sıvı arkasında bir hava bolus ile akışkan 50 ul ihtiva eden bir 0.5 ml şırıngaya PE10 boru bağlayın. 0.5 cm için ses tellerinin dikkatle PE boru ilerlemek ve yavaş yavaş solunum içine boşaltın. Bununla birlikte, alt solunum yolu için zaman zaman travma meydana gelir uNumunenin daha teslim olmadan bu tekniği şarkı. </li></ol>

<ol>
	<li>Rockman, H. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Segregation+of+atrial-specific+and+inducible+expression+of+an+atrial+natriuretic+factor+transgene+in+an+in+vivo+murine+model+of+cardiac+hypertrophy.">Segregation of atrial-specific and inducible expression of an atrial natriuretic factor transgene in an in vivo murine model of cardiac hypertrophy.</a> Proc. natl. Acad. Sci. U.S.A. 88, 8277-8281 (1991).</li><li>Watanabe, A., Hashimoto, Y., Ochiai, E., Sato, A., Kamei, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simple+method+for+confirming+correct+endotracheal+intubation+in+mice.">A simple method for confirming correct endotracheal intubation in mice.</a> Lab. Anim. 43, 399-401 (2009).</li><li>Orebaugh, S. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Succinylcholine:+adverse+effects+and+alternatives+in+emergency+medicine.">Succinylcholine: adverse effects and alternatives in emergency medicine.</a> Am. J. Emerg. Med. 17, 715-721 (1999).</li><li>Hastings, R. H., Summers-Torres, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+Laryngoscopy+in+Mice.">Direct Laryngoscopy in Mice.</a> Contemp. Lab. Anim. Sci. 38, 33-35 (1999).</li><li>Berul, C. I., Aronovitz, M. J., Wang, P. J., Mendelsohn, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+cardiac+electrophysiology+studies+in+the+mouse.">In vivo cardiac electrophysiology studies in the mouse.</a> Circulation. 94, 2641-2648 (1996).</li><li>Ewart, S. L., Gavett, S. H., Margolick, J., Wills-Karp, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cyclosporin+A+attenuates+genetic+airway+hyperresponsiveness+in+mice+but+not+through+inhibition+of+CD4++or+CD8++T+cells.">Cyclosporin A attenuates genetic airway hyperresponsiveness in mice but not through inhibition of CD4+ or CD8+ T cells.</a> Am. J. Respir. Cell Mol. Biol. 14, 627-634 (1996).</li><li>Ewart, S. L., Mitzner, W., DiSilvestre, D. A., Meyers, D. A., Levitt, R. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Airway+hyperresponsiveness+to+acetylcholine:+segregation+analysis+and+evidence+for+linkage+to+murine+chromosome+6.">Airway hyperresponsiveness to acetylcholine: segregation analysis and evidence for linkage to murine chromosome 6.</a> Am. J. Respir. Cell Mol. Biol. 14, 487-495 (1996).</li><li>Spoelstra, E. N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+and+simple+method+for+endotracheal+intubation+of+mice.">A novel and simple method for endotracheal intubation of mice.</a> Lab. 41, 128-135 (2007).</li><li>Boll, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-speed+single-breath-hold+micro-computed+tomography+of+thoracic+and+abdominal+structures+in+mice+using+a+simplified+method+for+intubation.">High-speed single-breath-hold micro-computed tomography of thoracic and abdominal structures in mice using a simplified method for intubation.</a> J. Comput. Assist. Tomogr. 34, 783-790 (2010).</li><li>Brown, R. H., Walters, D. M., Greenberg, R. S., Mitzner, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+method+of+endotracheal+intubation+and+pulmonary+functional+assessment+for+repeated+studies+in+mice.">A method of endotracheal intubation and pulmonary functional assessment for repeated studies in mice.</a> J. Appl. Physiol. 87, 2362-2365 (1999).</li><li>Vergari, A., Polito, A., Musumeci, M., Palazzesi, S., Marano, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Video-assisted+orotracheal+intubation+in+mice.">Video-assisted orotracheal intubation in mice.</a> Lab. Anim. 37, 204-206 (2003).</li><li>Vergari, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+method+of+orotracheal+intubation+in+mice.">A new method of orotracheal intubation in mice.</a> Eur. Rev. Med. Pharmacol. Sci. 8, 103-106 (2004).</li><li>Hamacher, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microscopic+wire+guide-based+orotracheal+mouse+intubation:+description,+evaluation+and+comparison+with+transillumination.">Microscopic wire guide-based orotracheal mouse intubation: description, evaluation and comparison with transillumination.</a> Lab. Anim. 42, 222-230 (2008).</li><li>Rivera, B., Miller, S., Brown, E., Price, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+method+for+endotracheal+intubation+of+mice+and+rats+used+in+imaging+studies.">A novel method for endotracheal intubation of mice and rats used in imaging studies.</a> Cont. Top. Lab. Anim. Sci. 44, 52-55 (2005).</li><li>MacDonald, K. D., Chang, H. Y., Mitzner, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+improved+simple+method+of+mouse+lung+intubation.">An improved simple method of mouse lung intubation.</a> J. Appl. Physiol. 106, 984-987 (2009).</li><li>De Vleeschauwer, S. p. I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Repeated+invasive+lung+function+measurements+in+intubated+mice:+an+approach+for+longitudinal+lung+research.">Repeated invasive lung function measurements in intubated mice: an approach for longitudinal lung research.</a> Lab. Anim. 45, 81-89 (2011).</li><li>Zhao, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+technique+for+retrograde+intubation+in+mice.">A technique for retrograde intubation in mice.</a> Lab. Anim. 35, 39-42 (2006).</li><li>Singer, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Left-sided+mouse+intubation:+description+and+evaluation.">Left-sided mouse intubation: description and evaluation.</a> Exp. Lung Res. 36, 25-30 (2010).</li></ol>

Yazarlar, hiçbir rakip mali çıkarlarını olmadığını beyan ederim.

Bu yazıda travmatik olmayan ve aynı hayvanda tekrar tekrar kullanılabilir fareler entübe için basit, güvenilir bir tekniği açıklar. Bu teknik bir mütevazı toplamı için satın alınabilir basit laboratuar ya da tıbbi ekipman ile gerçekleştirilebilir. Ilk Hastings ve arkadaşları tarafından rapor edilen 4 doğrudan laringoskopide tekniği, çeşitli amaçlar için de kullanılabilir, ama esas olarak doğru bir alt solunum sistemine test maddesi sunmak için. Biz nedeniyle tekrarlanan ölçüml...

Laboratuvar faresi biyoloji ve patobiyolojisinde başlıca memeli model olarak hemen hemen tüm türlerin yerini almıştır. Laboratuvar faresi açıkça ve yoğun insan hastalığının bir model olarak değer olduğu gösterilmiştir ve insan biyolojisi ve hastalık anlayışımızın gelişmeler paha biçilmez kanıtlamıştır küçük memeli türüdür. Kısa gebelik süresi ve önemli ölçüde daha düşük maliyetli bir biyomedikal araştırmalarda yaygın araç olarak boş ve transjenik farelerin geliştirme ve çalışma sağladı. Ancak, ortalama laboratuvar faresi (20-25 g) büyüklüğü dolayısıyla, bazı araştırmacılar, daha büyük memeli türler olan, fizyolojik ya da cerrahi araştırmalarda kendi çalışma sınırlı gelmiştir. Bu çalışmalarda fareler kullanarak bir engel derin anestezi altında fizyolojik ölçümler veya geniş cerrahi prosedürler sağlayacak entübasyon teknikleri ile karşılaşılan zorluk olduğunu. Trakeostomi 1, standart te olarak kullanılmaktadırçünkü gerekli bu tekniği ve mütevazı beceri yerine daha kolay yerine entübasyon NİK. Ancak, trakeostomi kronik veya kurtarma cerrahi çalışmalar için elverişli değildir; bu nedenle, akut deneyler ile sınırlıdır. Trakeostomi de iltihap veya hassas fizyolojik refleksleri önemli olduğu araştırma karıştırıcı bir değişken olabilir. Bizim laboratuvar diğer araştırmacılar tarafından açıklanan tekniklerin çoğunu denedim ve çeşitli nedenlerle için onları yetersiz buldu. Trakeostomi çok travmatik ve kanama ve hava yolu iltihabı neden olur. Çok daha sorunlu bunun fizibil tekrar edilemez olmasıdır. Ekipman mütevazı bir yatırım gerektiren birçok nispeten non-invaziv teknikler yeterince güvenilir değildir. Diğer teknikler ekipman belirli bir uygulamada çalışacak olmadığını bilmeden haklı zordur pahalı ekipman gerektirir. Böylece, biz artık t gerekli bir nontravmatik tekniğini geliştirmek için çalıştıhan özel bir ekipman mütevazı bir yatırım, hızlı bir şekilde gerçekleştirilebilir olabilir ve güvenilir bir şekilde, kronik modellerinde tekrar olabilir ve hayvanların çok sayıda kullanılabilir. İşte biz böyle bir teknik rapor.

<table border="0" cellpadding="0" cellspacing="0" width="706">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="17">
			<td height="17" style="height:17px;width:411px;">Operating Otoscope Head</td>
			<td style="width:144px;">Welch Allyn</td>
			<td align="right" style="width:83px;">21700</td>
			<td style="width:69px;">&#160;$188.98</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Otoscope Handle</td>
			<td>Welch Allyn</td>
			<td align="right">71000</td>
			<td>&#160;$112.20</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Reuseable Speculum</td>
			<td>Welch Allyn</td>
			<td align="right">22002</td>
			<td>&#160;$ 3.98</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Fine Forceps</td>
			<td>Miltex</td>
			<td>18-779</td>
			<td>&#160;$107.18</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Small clamp stand to hold otoscope</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;"></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Optional Equipment</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Isoflurane Vaporizer (multiple circuit with vacuum waste gas handling)</td>
			<td>Summit Medical</td>
			<td></td>
			<td align="right">$3,000</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Flexivent (Animal Ventilator)</td>
			<td>SCIREQ</td>
			<td></td>
			<td align="right">$35,000</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;"></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;"></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Supplies</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Intravenous catheter (20 ga x 1 inch)</td>
			<td>BD</td>
			<td align="right">381233</td>
			<td>&#160;$&#160; 9.62</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Polyethylene tubing (PE10) (0.011 inch I.D., 0.024 inch O.D.) 100 ft</td>
			<td>Intramedic, Clay-Adams</td>
			<td align="right">427401</td>
			<td>&#160;$115.70</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Ketamine 100 mg/ml (10 ml bottle)</td>
			<td>Butler</td>
			<td align="right">23061</td>
			<td>&#160;$10.00</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Xylazine (100 ml bottle)</td>
			<td>Vedco</td>
			<td align="right">24105</td>
			<td>&#160;$20.00</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Isoflurane (250 ml bottle)</td>
			<td></td>
			<td></td>
			<td>&#160;$15.00</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">vecuronium bromide 10 mg/10 ml</td>
			<td>Pfizer</td>
			<td>NDC 0069-0094-01</td>
			<td align="right">$15.00</td>
		</tr>
	</tbody>
</table>

endotracheal intubation in mice via direct laryngoscopy using an otoscope

Fareler, vahşi tipli ve transjenik hem şu anda, biyomedikal araştırmalarda asıl memeli modeli vardır. Entübasyon ve mekanik ventilasyon derin anestezi veya akciğer fonksiyon ölçümleri altında ameliyat gerektiren bütün hayvan deneyleri için gereklidir. Trakeostomi mekanik havalandırma sağlamak için bu farelerde entübasyon için standart olmuştur. Orotrakeal entübasyon nedeniyle önemli teknik zorluk veya çok özel ve pahalı ekipman için bir gereklilik rapor edilmiştir ancak başarılı birçok çalışmada kullanılmıştır olmamıştır. Burada bir 2.0 mm spekulum ile donatılmış bir otoskop kullanarak ve bir endotrakeal tüp olarak 20 G intravenöz kateter kullanarak doğrudan laringoskopi bir teknik rapor. Biz entübe ve farelerde akciğer fonksiyonunun doğru değerlendirmeler yapmak için yaygın ve güvenilir bu tekniği kullandık. Bu teknik, deneyimli ellerde aslında hiçbir hayvan kaybı ile güvenli kanıtlamıştır. Ayrıca, bu teknik,Kronik bir model farelerin tekrarlanan çalışmalar için de kullanılabilir.

Yukarıdaki tekniği ile entübasyon, güvenilir ve hızlı. Endotrakeal tüpün uygun yerleştirme en kolay ve batık ekspiratuar uzuv süresi dolan gaz kabarcıklarının oluşması gözlemleyerek tarafından doğrulanmadı ventilatör devresi (genellikle bir PEEP tuzak) ve (Şekil 1) izleme, bir hava yolu basıncı üzerindeki olumsuz sapmaların. Hava yolu basıncı izleme üzerinde olumsuz deplasmanlar en güvenilir. Diğer ekspiratuar vantilatör devresi 2&#39;ye bağlı intravenöz bor...

Watch this Scientific Journal Video about Endotracheal Intubation in Mice via Direct Laryngoscopy Using an Otoscope at JoVE.com

Endotracheal Intubation in Mice via Direct Laryngoscopy Using an Otoscope

Biz 2.0 mm spekulum ile bir otoskop kullanarak direkt laringoskopi yoluyla farelerde entübasyon için, basit, güvenilir ve nispeten ucuz bir yöntem geliştirdik. Bu teknik, atravmatik ve kronik deneylerde tekrarlanan ölçümler için de kullanılabilir. Biz üstün trakeostomi veya daha önce cerrahi dışı teknikler bildirdi bulabilirsiniz.

Farelerde Endotrakeal Entübasyon<em&gt; Via</emBir Otoskop Kullanma&gt; Doğrudan Larengoskopi

Mice, both wildtype and transgenic, are the principal mammalian model in biomedical research currently. Intubation and mechanical ventilation are ...

endotracheal-intubation-mice-via-direct-laryngoscopy-using-an

Research

JoVE Journal

Medicine

Endotracheal Intubation in Mice via Direct Laryngoscopy Using an Otoscope

30.9K Görüntüleme.  VA San Diego Healthcare System.  Biz 2.0 mm spekulum ile bir otoskop kullanarak direkt laringoskopi yoluyla farelerde entübasyon için, basit, güvenilir ve nispeten ucuz bir yöntem geliştirdik. Bu teknik, atravmatik ve kronik deneylerde tekrarlanan ölçümler için de kullanılabilir. Biz üstün trakeostomi veya daha önce cerrahi dışı teknikler bildirdi bulabilirsiniz. 

Video: Endotracheal Intubation in Mice via Direct Laryngoscopy Using an Otoscope

Using Quantitative Real-time PCR to Determine Donor Cell Engraftment in a Competitive Murine Bone Marrow Transplantation Model

Murine bone marrow transplantation models provide an important tool in measuring hematopoietic stem cell (HSC) functions and determining genes/molecules that regulate HSCs. In these transplant model systems, the function of HSCs is determined by the ability of these cells to engraft and reconstitute lethally irradiated recipient mice. Commonly, the donor cell contribution/engraftment is measured by antibodies to donor- specific cell surface proteins using flow cytometry. However, this method heavily depends on the specificity and the ability of the cell surface marker to differentiate donor-derived cells from recipient-originated cells, which may not be available for all mouse strains. Considering the various backgrounds of genetically modified mouse strains in the market, this cell surface/ flow cytometry-based method has significant limitations especially in mouse strains that lack well-defined surface markers to separate donor cells from congenic recipient cells. Here, we reported a PCR-based technique to determine donor cell engraftment/contribution in transplant recipient mice. We transplanted male donor bone marrow HSCs to lethally irradiated congenic female mice. Peripheral blood samples were collected at different time points post transplantation. Bone marrow samples were obtained at the end of the experiments. Genomic DNA was isolated and the Y chromosome specific gene, Zfy1, was amplified using quantitative Real time PCR. The engraftment of male donor-derived cells in the female recipient mice was calculated against standard curve with known percentage of male vs. female DNAs. Bcl2 was used as a reference gene to normalize the total DNA amount. Our data suggested that this approach reliably determines donor cell engraftment and provides a useful, yet simple method in measuring hematopoietic cell reconstitution in murine bone marrow transplantation models. Our method can be routinely performed in most laboratories because no costly equipment such as flow cytometry is required.

Determining donor cell engraftment presents a challenge in mouse bone marrow transplant models that lack well-defined phenotypical markers. We described a methodology to quantify male donor cell engraftment in female transplant recipient mice. This method can be used in all mouse strains for the study of HSC functions.

Rekabetçi bir mürin Kemik İliği Transplantasyon Modelinde Donör Hücre Aşı belirleme Kantitatif Gerçek-zamanlı PCR kullanarak

Murine bone marrow transplantation models provide an important tool in measuring hematopoietic stem cell (HSC) functions and determining genes/molecules ...

Tıp

A Simple Method of Mouse Lung Intubation

A simple procedure to intubate mice for pulmonary function measurements would have several advantages in longitudinal studies with limited numbers or expensive animal. One of the reasons that this is not done more routinely is that it is relatively difficult, despite there being several published studies that describe ways to achieve it. In this paper we demonstrate a procedure that eliminates one of the major hurdles associated with this intubation, that of visualizing the trachea during the entire time of intubation. The approach uses a 0.5 mm fiberoptic light source that serves as an introducer to direct the intubation cannula into the mouse trachea. We show that it is possible to use this procedure to measure lung mechanics in individual mice over a time course of at least several weeks. The technique can be set up with relatively little expense and expertise, and it can be routinely accomplished with relatively little training. This should make it possible for any laboratory to routinely carry out this intubation, thereby allowing longitudinal studies in individual mice, thereby minimizing the number of mice needed and increasing the statistical power by using each mouse as its own control.

This paper describes a striaghforward and efficient method of intubating mice for pulmonary function measurements or pulmonary instillation, that allows the mice to recover and be studied at later times. The procedure involves an inexpensive fiberoptic light source that directly illuminates the trachea.

Fare Akciğer Entübasyonun Basit Bir Yöntem

A  simple procedure to intubate mice for pulmonary function measurements would  have several advantages in longitudinal studies with limited numbers or  ...

Heterotopic Mucosal Engrafting Procedure for Direct Drug Delivery to the Brain in Mice

Delivery of therapeutics into the brain is impeded by the presence of the blood-brain barrier (BBB) which restricts the passage of polar and high molecular weight compounds from the bloodstream and into brain tissue. Some direct delivery success in humans has been achieved via implantation of transcranial catheters; however this method is highly invasive and associated with numerous complications. A less invasive alternative would be to dose the brain through a surgically implanted, semipermeable membrane such as the nasal mucosa that is used to repair skull base defects following endoscopic transnasal tumor removal surgery in humans. Drug transfer though this membrane would effectively bypass the BBB and diffuse directly into the brain and cerebrospinal fluid. Inspired by this approach, a surgical approach in mice was developed that uses a donor septal mucosal membrane engrafted over an extracranial surgical BBB defect. This model has been shown to effectively allow the passage of high molecular weight compounds into the brain. Since numerous drug candidates are incapable of crossing the BBB, this model is valuable for performing preclinical testing of novel therapies for neurological and psychiatric diseases.

A mouse model of human endoscopic skull base reconstruction has been developed that creates a semipermeable interface between the brain and nose using nasal mucosal grafts. This method allows researchers to study delivery to the central nervous system of high molecular weight therapeutics which are otherwise excluded by the blood-brain barrier when administered systemically.

Farelerde Beyin Doğrudan İlaç Teslim heterotopik mukozal aşılanması Prosedürü

Delivery of therapeutics into the brain is impeded by the presence of the blood-brain barrier (BBB) which restricts the passage of polar and high ...

An Improved Method for Rapid Intubation of the Trachea in Mice

Despite some anatomical and physiological differences, mouse models continue to be an essential tool for studying human lung disease. Bleomycin toxicity is a commonly used model to study both acute lung injury and fibrosis, and multiple methods have been developed for administering bleomycin (and other toxic agents) into the lungs. However, many of these approaches, such as transtracheal instillation, have inherent drawbacks, including the need for strong anesthetics and survival surgery. This paper reports a quick, reproducible method of intratracheal intubation that involves mild inhaled anesthesia, visualization of the trachea, and the use of a surrogate spirometer to confirm exposure. As a proof of concept, 8-12 week old C57BL/6 mice were administered either 2.0 U/kg of bleomycin or an equivalent volume of PBS, and both damage and fibrotic endpoints were measured post-exposure. This procedure allows researchers to treat a large cohort of mice in a relatively short period with little expense and minimal post-procedure care.

This article presents a rapid and simple method for administering bleomycin directly into the mouse trachea via intubation. Key advantages of this method are that it is highly reproducible, easy to master, and does not require specialized equipment or lengthy recovery times.

Farelerde Trakea hızlı intübasyon için geliştirilmiş bir yöntem

Despite some anatomical and physiological differences, mouse models continue to be an essential tool for studying human lung disease. Bleomycin toxicity ...

Inducing Acute Lung Injury in Mice by Direct Intratracheal Lipopolysaccharide Instillation

Airway administration of lipopolysaccharide (LPS) is a common way to study pulmonary inflammation and acute lung injury (ALI) in small animal models. Various approaches have been described, such as the inhalation of aerosolized LPS as well as nasal or intratracheal instillation. The presented protocol describes a detailed step-by-step procedure to induce ALI in mice by direct intratracheal LPS instillation and perform FACS analysis of blood samples, bronchoalveolar lavage (BAL) fluid, and lung tissue. After intraperitoneal sedation, the trachea is exposed and LPS is administered via a 22 G venous catheter. A robust and reproducible inflammatory reaction with leukocyte invasion, upregulation of proinflammatory cytokines, and disruption of the alveolo-capillary barrier is induced within hours to days, depending on the LPS dosage used. Collection of blood samples, BAL fluid, and lung harvesting, as well as the processing for FACS analysis, are described in detail in the protocol. Although the use of the sterile LPS is not suitable to study pharmacologic interventions in infectious diseases, the described approach offers minimal invasiveness, simple handling, and good reproducibility to answer mechanistic immunological questions. Furthermore, dose titration as well as the use of alternative LPS preparations or mouse strains allow modulation of the clinical effects, which can exhibit different degrees of ALI severity or early vs. late onset of disease symptoms.

Presented here is a step-by-step procedure to induce acute lung injury in mice by direct intratracheal lipopolysaccharide instillation and to perform FACS analysis of blood samples, bronchoalveolar lavage fluid, and lung tissue. Minimal invasiveness, simple handling, good reproducibility, and titration of disease severity are advantages of this approach.

Doğrudan Intratracheal Lipopolysaccharide Instilasyon ile farelerde akut akciğer hasarının İndükleme

Airway administration of lipopolysaccharide (LPS) is a common way to study pulmonary inflammation and acute lung injury (ALI) in small animal models. ...

Repeated Orotracheal Intubation in Mice

The literature describes several methods for mouse intubation that either require visualization of the glottis through the oral cavity or incision in the ventral neck for direct confirmation of cannula placement in the trachea. The relative difficulty or the tissue trauma induced to the subject by such procedures can be an impediment to an investigator&#8217;s ability to perform longitudinal studies. This article illustrates a technique in which physical manipulation of the mouse following the use of a depilatory to remove hair from the ventral neck permits transcutaneous visualization of the trachea for orotracheal intubation regardless of degree of skin pigmentation. This method is innocuous to the subject and easily achieved with a limited understanding of murine anatomy. This refined approach facilitates repeated intubation, which may be necessary for monitoring progression of disease or instillation of treatments. Using this method may result in a reduction of the number of animals and technical skill required to measure lung function in mouse models of respiratory disease.

The goal of this article is to describe a refined method of intubation of the laboratory mouse. The method is noninvasive and, therefore, ideal for studies that require serial monitoring of respiratory function and/or instillation of treatments into the lung.

Farelerde Tekrarlanan Orotracheal Entübasyon

The literature describes several methods for mouse intubation that either require visualization of the glottis through the oral cavity or incision in the ...

Blind Endotracheal Intubation in Neonatal Rabbits

The newborn rabbit is a useful animal model for various pathologies and procedures. Airway management of the rabbit is complex due to its anatomical characteristics, which is further complicated in the case of the newborn. Of the different methods of advanced airway management, endotracheal intubation is less aggressive than tracheostomy, and is more feasible than supraglottic management given the lack of supraglottic devices of such a small size. As direct glottis visualization is very difficult in animals this size, this blind intubation model is presented as an effective alternative, especially for experiments requiring prolonged anesthesia. Using this method, we performed blind intubations with a 90% success rate.

We describe a technique of endotracheal intubation in newborn rabbits after esophageal catheterization with a gastric tube.

Yenidoğan Tavşanlarında Kör Endotrakeal Entübasyon

The newborn rabbit is a useful animal model for various pathologies and procedures. Airway management of the rabbit is complex due to its anatomical ...

Direct Drug Delivery to Kidney via the Renal Artery

There is a need for directed injections to enable increased and specific renal exposure for efficient evaluation of drug targets in the renal research field. Accumulation of drugs in certain organs may give rise to adverse and unwanted effects, depending on the nature of injectate. To minimize spillover and/or accumulation in other tissues, the herein described method directs the formulation into the renal artery bloodstream by inserting a catheter in the infra renal aorta, just below where it branches into the renal artery, resulting in the kidney as first reached organ and distributing of formulation throughout the kidney.
This manuscript provides a detailed description of the method, as well as its challenges and difficulties. It guides the experimenter to become skillful with this type of microsurgery that requires accuracy under sterile conditions. Speed is crucial for minimizing the ischemia and practicing the procedure will increase the chance of successful injections without adverse effects. By modulating the time between injection and reperfusion as well as the injected volume, the risk of spillover to other organs is mitigated. 
Note that this technique is suitable for single dosing strategies.

This manuscript describes a method for targeted delivery to a single kidney via a catheter placed in the infrarenal abdominal aorta in the mouse.

Böbrek Atardamarı Ile Böbreğe Doğrudan İlaç Verilmesi

There is a need for directed injections to enable increased and specific renal exposure for efficient evaluation of drug targets in the renal research ...

Endotracheal Intubation Using a Flexible Intubation Endoscope as a Standardized Model for Safe Airway Management in Swine

Endotracheal intubation is often a basic requirement for translational research in porcine models for various interventions that require a secured airway or high ventilation pressures. Endotracheal intubation is a challenging skill,&#160;requiring a minimum number of successful endotracheal intubations to achieve a high success rate under optimal conditions, which is often unachievable for non-anaesthesiology researchers. Due to the specific porcine airway anatomy, a difficult airway can usually be assumed. The impossibility of establishing a secure airway can result in injury, adverse events, or death of the laboratory animal. Using a prospective, randomized, controlled evaluation approach, it has been shown that fiberoptic-assisted endotracheal intubation takes longer but has a higher first-pass success rate than conventional intubation without causing clinically relevant drops in oxygen saturation. This model presents a standardized regimen for endoscopically guided endotracheal intubation, providing a secured airway, especially for researchers who are inexperienced in the technique of endotracheal intubation via direct laryngoscopy. This procedure is expected to minimize animal suffering and unnecessary animal losses.

The use of pigs in research has increased in recent years. Nevertheless, pigs are characterized by difficult airway anatomy. By demonstrating how to perform endoscopically guided endotracheal intubation, the present protocol aims to further increase laboratory animals' safety to avoid animal suffering and unnecessary death.

Domuzlarda Güvenli Hava Yolu Yönetimi için Standartlaştırılmış Bir Model Olarak Esnek Entübasyon Endoskobu Kullanılarak Endotrakeal Entübasyon

Endotracheal intubation is often a basic requirement for translational research in porcine models for various interventions that require a secured airway ...

<meta charset="utf8"></head><body>Hayatta Kalamayan Sıçan Modelleri için Trakeotomi Tabanlı Endotrakeal Entübasyon Protokolü

Endotracheal Intubation via Tracheotomy and Subsequent Thoracotomy in Rats for Non-Survival Applications

Endotracheal intubation and subsequent ventilation are often basic requirements for translational research in rat models for various interventions that require controlled or high ventilation pressures or access to the thoracic cavity and organs. Conventional endoorotracheal intubation using the anatomically existing route through the mouth is well suited for survival experiments. However, this procedure poses some challenges, including generally higher levels of the required experience and technical skill, more advanced equipment, and greater time effort with relevant intubation failure rates and complications such as tracheal perforation, temporary systemic hypooxygenation, and relevant aerial leakage.
This manuscript, therefore, presents a detailed step-by-step protocol for endotracheal intubation through tracheotomy in non-survival rat models when guaranteed intubation success, constant oxygenation levels, high ventilation pressures, or open thoracotomy are required.
The protocol emphasizes the importance of meticulous surgical technique to ensure consistent and reliable outcomes, especially for researchers who are inexperienced or lack routine in the technique of endoorotracheal intubation via direct laryngoscopy. This procedure is, therefore, expected to minimize animal suffering and unnecessary animal losses.

<meta charset="utf8"></head><body>Hayatta kalamayan uygulamalar için sıçanlarda trakeotomi ile endotrakeal entübasyon ve ardından torakotomi

Here we introduce a standardized procedure for endotracheal intubation via tracheotomy followed by thoracotomy in rats, aimed at enhancing the precision and reproducibility of non-survival applications requiring invasive ventilation and exposure of thoracic organs in in vivo rat models.

Hayatta kalamayan uygulamalar için sıçanlarda trakeotomi ile endotrakeal entübasyon ve ardından torakotomi

Endotracheal intubation and subsequent ventilation are often basic requirements for translational research in rat models for various interventions that ...