Los autores desean agradecer a la División de Transmitidas por Vectores de Enfermedades Animales de Recursos Rama, específicamente Andrea Peterson, Lisa Massoudi, O&#39;Brien Verna y Juan Liddell para su cuidado y mantenimiento de los ratones y conejos. También nos gustaría dar las gracias a Amy Ullmann, Russell Teresa, y Barbara J. Johnson por su contribución hacia este manuscrito. Por último, queremos agradecer a Alissa Eckert en la Oficina del Director Adjunto de Comunicación de los CDC para la producción de las ilustraciones gráficas y Lavelle Judy para dirigir todos los aspectos legales relacionados con la filmación de este manuscrito.

1. Hemolinfa colección para la preparación de diapositivas (Película 1) <ol><li> Suavemente eliminar activamente las garrapatas de alimentación de un animal y el lugar en el peróxido de hidrógeno al 3% tópica durante 5 minutos y luego en etanol al 70% durante 10 minutos para esterilizar la superficie. </li><li> Con una pluma de Papanicolaou dibujar un círculo sobre un portaobjetos recubierta de silano y coloque la garrapata dentro del círculo de la pluma de Papanicolaou. <ol><li> Silano portaobjetos revestidos se utilizan para la mejor adherencia de la hemolinfa al portaobjetos de microscopio. </li></ol></li><li> Ver la garrapata con un microscopio de disección (1X objetivo, ocular 10X, la magnificación de 3.5X). </li><li> Empuje suavemente hacia abajo en el dorso de la garrapata con unas pinzas de splay patas de la garrapata y la inmovilización de la garrapata. </li></ol> Nota: Cuando la inmovilización de la garrapata no presionar demasiado fuerte porque esto puede afectar el intestino o la punción de la garrapata y contaminar la hemolinfa. <ol start="5"><li> Amputar la pierna de la garrapata o las patas ºe distal conjunta con una punta fina desechables bisturí. Para determinar la infectividad a través de la hemolinfa sólo una pierna tiene que ser amputada. Para la recolección de la hemolinfa en un portaobjetos de varios tramos puede ser amputada. </li></ol> Nota: No corte la pierna demasiado cerca del cuerpo, porque esto puede causar la contaminación del intestino medio de la hemolinfa. <ol start="6"><li> Después de la pierna o piernas se cortan seguir aplicando presión suavemente a dorso de la garrapata de la hemolinfa de secretar fuera de las patas en la diapositiva. Suavemente mueva alrededor de la garrapata en la diapositiva para difundir la hemolinfa. </li></ol> 2. Extirpación de la glándula salival (Cine 2 y 3) <ol><li> Spot varias piscinas de 25 l de tampón fosfato salino (PBS) sobre un portaobjetos de microscopio y se coloca una marca en una de las piscinas de PBS. </li><li> Ver la garrapata con un microscopio de disección (1X objetivo, ocular 10X, la magnificación de 3.5X). </li><li> Estabilizar la garrapata con pinza de punta finas manteniendo la cabezuela base (partes de la boca) o la parte trasera de la garrapata. </li><li> Inserte las pinzas de punta fina en la parte posterior de la garrapata y el tramo hasta que la garrapata es el dorso de exponer los órganos. Si se desea, el intestino medio puede ser eliminado en este momento y se transfiere a una piscina fresca de PBS en un portaobjetos de microscopio o de un tubo de microcentrífuga que contiene PBS. </li><li> Encuentra el par de glándulas salivales (como racimos de uva), ubicado al lado de las piernas bilateral de la garrapata. Si las glándulas salivales no son visibles entre los escombros garrapata mover la parte principal de la garrapata, que todavía contiene las glándulas salivales, a un grupo fresco de PBS para reducir los trastornos y pérdida de las glándulas salivales. </li></ol> Nota: A principios de la alimentación, las glándulas salivales son más difíciles de localizar porque no están tan desarrolladas como en comparación con el más adelante en la alimentación. <ol start="6"><li> Retire las glándulas salivales de la garrapata con unas pinzas de punta fina y el lugar en una piscina fresca de PBS. </li><li> Wiª de punta fina pinza de transferencia de las glándulas salivales a otra limpia 25 l grupo PBS, repita este paso de lavado 3-4 veces más para eliminar los microorganismos externos y los restos de garrapata. </li></ol> Nota: Lavar las agrupaciones de las glándulas salivales suavemente para reducir la pérdida y alteración de las glándulas salivales individuales. <ol start="8"><li> Colocar las glándulas en una piscina limpia de PBS en un portaobjetos recubierta de silano o en un tubo de microcentrífuga que contiene PBS. </li></ol> 3. La saliva recogida (Película 4) <ol><li> Retire con cuidado garrapatas hembras adultas de conejo o de otro host usando unas pinzas finas con punta justo antes de que caen apagado totalmente llena de sangre, días aproximadamente 5-7 después de la fijación. </li><li> Pegue la garrapata cerca de hinchada en un extremo de un portaobjetos con cinta adhesiva. La cinta debe estar colocada aproximadamente a ¾ de su altura dorso de la garrapata hacia la cabeza, dejando cóndilo de la garrapata base (partes de la boca) expuestos.</li><li> Cuando la cinta se encuentra con el borde anterior de la superficie dorsal garrapatas pipeta 5 ml de solución de pilocarpina 5% (en metanol). Permitir la cinta para absorber la pilocarpina sobre el dorso de garrapata, sin permitir que la pilocarpina para entrar en contacto con cabezuela base de la garrapata. </li><li> Montar un trozo de plastilina no tóxica en el portaobjetos de un microscopio de aproximadamente una pulgada de piezas bucales de la garrapata. </li><li> Utilizando unas pinzas de punta fina rompa la punta de un tubo capilar sacó 28 al diámetro deseado. </li><li> Ver cóndilo de la garrapata base bajo un microscopio de disección. </li><li> Suavemente encajar hipostoma de la garrapata en el tubo capilar sacó permitiendo que los palpos maxilares para residir en el exterior del tubo capilar. </li><li> Presione el extremo opuesto del tubo capilar en la arcilla de modelado para mantener el tubo capilar en su lugar. </li><li> Coloque la garrapata montada la boca agua dentro de una cámara oscura con una alta humedad (por ejemplo, una caja de poliestireno con tapa forrada con pap húmeda<html> er toallas). Al girar el portaobjetos para que los puntos hipostoma a la parte inferior del recipiente, permitiendo que la gravedad para ayudar en la recogida de la saliva. </li><li> Poner el recipiente a temperatura ambiente. </li><li> Vigilar de cerca las garrapatas salivando durante la primera hora, y recoger la saliva como se genera mediante la expulsión fuera de los tubos capilares con un bulbo de pipeta de Pasteur. Después de la primera hora, visita acumulación cada hora durante al menos 4 horas. Continuar para recoger la saliva, ya que se genera. </li></ol> Nota: la adquisición de la saliva puede detener una vez suficiente saliva se recoge en el estudio que se realiza. <ol start="12"><li> Si las garrapatas no están salivando o si se requiere más saliva, salivación en ocasiones puede ser inducida mediante el tubo capilar para masajear el hipostoma. </li><li> Añadir 0,1 volúmenes de cóctel inhibidor de la proteasa a la saliva y almacenar a -80 ° C hasta que se necesite. </li></ol> 4. Los resultados representativos e_content &quot;Película&gt; 1 muestra cómo llevar a cabo una parte alimenta la ninfa I. scapularis y amputar las piernas para recoger la hemolinfa en un portaobjetos de microscopio. Una vez que la pierna o piernas son amputadas de un líquido claro es secretada (Figura 1A y 1B). Si el intestino medio se rompe la hemolinfa aparece turbia como lo es sale de la pierna amputada (s) (Figura 1C y 1D). La extracción de las glándulas salivares después de la ninfa ha estado alimentando durante 48 ó 72 horas se demuestra en las películas 2 y 3. Después de la garrapata es perforado en general hay una gran cantidad de desechos (que consta de tráquea, malpigiana túbulos, sangre, tejido conjuntivo, etc), para evitar la pérdida o alteración de las glándulas salivales mover la garrapata a una piscina fresca de PBS. Las figuras 2A y 2B muestran en las glándulas salivales se encuentran después de la ninfa ha sido abierto y 2C figura muestra eliminado grupos de glándulas salivales en un charco de PBS. La obtención de saliva crearse a partir de I. las hembras adultas scapularis i<html> s se muestra en la película de 4 y la figura 3. Una garrapata salivando en un tubo capilar se observa en la película 5. Este método de extracción de saliva utiliza la pilocarpina para estimular la salivación y puede producir más de 20 l de saliva por la garrapata hembra adulta. <img alt="Figura 1." src="/files/ftp_upload/3894/3894fig1.jpg" /> Figura 1. Las estructuras con etiqueta de una ninfa I. scapularis. Película 1. Colección de Ixodes scapularis hemolinfa. <a href="http://www.jove.com/files/ftp_upload/3894/3894movie1.avi">Haga clic aquí para ver la película</a> . <img alt="Figura 2." src="/files/ftp_upload/3894/3894fig2.jpg" /> Figura 2. No contaminados (A y B) y contaminados (C + D) hemolinfa exudado de la pierna de la ninfa. Película 2. Extracción de las glándulas salivales de un 48 horas alimentados I. scapularis ninfa.pload/3894/3894movie2.avi &quot;&gt; Haga clic aquí para ver la película. Película 3. Extracción de las glándulas salivales de un 72 horas alimentados I. scapularis ninfa. <a href="http://www.jove.com/files/ftp_upload/3894/3894movie3.avi">Haga clic aquí para ver la película</a> . <img alt="Figura 3." src="/files/ftp_upload/3894/3894fig3.jpg" /> Figura 3. Glándulas salivales Ixodes scapularis ninfa. (A y B) Ejemplos de glándulas salivales en una ninfa de 72 horas alimentado, antes de la extracción. (C) Se ha eliminado del clúster glándula salival. Colección de películas 4. La saliva configurar de un adulto I. garrapata hembra scapularis. <a href="http://www.jove.com/files/ftp_upload/3894/3894movie4.avi">Haga clic aquí para ver la película</a> . <img alt="Figura 4." src="/files/ftp_upload/3894/3894fig4.jpg" /> Figura 4. Colección de saliva de un adulto I. scapularis t mujeresIcks. (A y B) Marque montado en una diapositiva con su hipostoma al final sacó del tubo capilar con el fin unpulled del tubo capilar en manos de plastilina. (C + D) humidificado cámara que contiene agua la boca adulta I. scapularis garrapatas hembras. Película 5. Ixodes scapularis garrapata hembra la boca agua en un tubo capilar. <a href="http://www.jove.com/files/ftp_upload/3894/3894movie5.avi">Haga clic aquí para ver la película</a> .

<ol>
	<li>Quach, K. A., Boctor, F. N., Elston, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=What's+eating+you?+Hyalomma+ticks.">What's eating you? Hyalomma ticks.</a> Cutis. 87, 165-167 (2011).</li><li>Graham, J., Stockley, K., Goldman, R. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tick-borne+illnesses:+a+CME+update.">Tick-borne illnesses: a CME update.</a> Pediatr. Emerg. Care. 27, 141-147 (2011).</li><li>Nuttall, P. A., Paesen, G. C., Lawrie, C. H., Wang, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vector-host+interactions+in+disease+transmission.">Vector-host interactions in disease transmission.</a> J. Mol. Microbiol. Biotechnol. 2, 381-386 (2000).</li><li>Estrada-Pena, A., Jongejan, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ticks+feeding+on+humans:+a+review+of+records+on+human-biting+Ixodoidea+with+special+reference+to+pathogen+transmission.">Ticks feeding on humans: a review of records on human-biting Ixodoidea with special reference to pathogen transmission.</a> Exp. Appl. Acarol. 23, 685-715 (1999).</li><li>Nuttall, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathogen-tick-host+interactions:+Borrelia+burgdorferi+and+TBE+virus.">Pathogen-tick-host interactions: Borrelia burgdorferi and TBE virus.</a> Zentralbl Bakteriol. 289, 492-505 (1999).</li><li>Jones, L. D., Hodgson, E., Nuttall, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enhancement+of+virus+transmission+by+tick+salivary+glands.">Enhancement of virus transmission by tick salivary glands.</a> J. Gen. Virol. 70, 1895-1898 (1989).</li><li>Labuda, M., Nuttall, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tick-borne+viruses.">Tick-borne viruses.</a> Parasitol. 129, 221-245 (2004).</li><li>Socolovschi, C., Mediannikov, O., Raoult, D., Parola, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Update+on+tick-borne+bacterial+diseases+in+Europe.">Update on tick-borne bacterial diseases in Europe.</a> Parasite. 16, 259-273 (2009).</li><li>Zhang, L., 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=Molecular+Interactions+that+Enable+Movement+of+the+Lyme+Disease+Agent+from+the+Tick+Gut+into+the+Hemolymph.">Molecular Interactions that Enable Movement of the Lyme Disease Agent from the Tick Gut into the Hemolymph.</a> PLoS Pathog. 7, e1002079 (2011).</li><li>Piesman, J., Schneider, B. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynamic+changes+in+Lyme+disease+spirochetes+during+transmission+by+nymphal+ticks.">Dynamic changes in Lyme disease spirochetes during transmission by nymphal ticks.</a> Exp. Appl. Acarol. 28, 141-145 (2002).</li><li>Brossard, M., Wikel, S. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tick+immunobiology.">Tick immunobiology.</a> Parasitol. , S161-S176 (2004).</li><li>Machackova, M., Obornik, M., Kopecky, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+salivary+gland+extract+from+Ixodes+ricinus+ticks+on+the+proliferation+of+Borrelia+burgdorferi+sensu+stricto+in+vivo.">Effect of salivary gland extract from Ixodes ricinus ticks on the proliferation of Borrelia burgdorferi sensu stricto in vivo.</a> Folia Parasitol. 53, 153-158 (2006).</li><li>Nuttall, P. A., Labuda, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tick-host+interactions:+saliva-activated+transmission.">Tick-host interactions: saliva-activated transmission.</a> Parasitol. 129, 177-189 (2004).</li><li>Anguita, J., Hedrick, M. N., Fikrig, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adaptation+of+Borrelia+burgdorferi+in+the+tick+and+the+mammalian+host.">Adaptation of Borrelia burgdorferi in the tick and the mammalian host.</a> FEMS Microbiol. Rev. 27, 493-504 (2003).</li><li>Hovius, J. W., van Dam, A. P., Fikrig, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tick-host-pathogen+interactions+in+Lyme+borreliosis.">Tick-host-pathogen interactions in Lyme borreliosis.</a> Trends Parasitol. 23, 434-438 (2007).</li><li>Dunham-Ems, S. M., 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=Live+imaging+reveals+a+biphasic+mode+of+dissemination+of+Borrelia+burgdorferi+within+ticks.">Live imaging reveals a biphasic mode of dissemination of Borrelia burgdorferi within ticks.</a> Journal Clin. Invest. 119, 3652-3665 (2009).</li><li>Ribeiro, J. M., Mather, T. N., Piesman, J., Spielman, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dissemination+and+salivary+delivery+of+Lyme+disease+spirochetes+in+vector+ticks+(Acari:+Ixodidae).">Dissemination and salivary delivery of Lyme disease spirochetes in vector ticks (Acari: Ixodidae).</a> J. Med. Entomol. 24, 201-205 (1987).</li><li>Piesman, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transmission+of+Lyme+disease+spirochetes+(Borrelia+burgdorferi.">Transmission of Lyme disease spirochetes (Borrelia burgdorferi.</a> Exp. Appl. Acarol. 7, 71-80 (1989).</li><li>De Silva, A. M., Fikrig, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Growth+and+migration+of+Borrelia+burgdorferi+in+Ixodes+ticks+during+blood+feeding.">Growth and migration of Borrelia burgdorferi in Ixodes ticks during blood feeding.</a> Am. J. Trop. Med. Hyg. 53, 397-404 (1995).</li><li>Horka, H., Cerna-Kyckova, K., Skallova, A., Kopecky, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tick+saliva+affects+both+proliferation+and+distribution+of+Borrelia+burgdorferi+spirochetes+in+mouse+organs+and+increases+transmission+of+spirochetes+to+ticks.">Tick saliva affects both proliferation and distribution of Borrelia burgdorferi spirochetes in mouse organs and increases transmission of spirochetes to ticks.</a> Int. J. Med. Microbiol. 299, 373-380 (2009).</li><li>Brossard, M., Wikel, S. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunology+of+interactions+between+ticks+and+hosts.">Immunology of interactions between ticks and hosts.</a> Med. Vet. Entomol. 11, 270-276 (1997).</li><li>Wikel, S. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tick+modulation+of+host+immunity:+an+important+factor+in+pathogen+transmission.">Tick modulation of host immunity: an important factor in pathogen transmission.</a> Int. J. Parasitol. 29 (99), 851-859 (1999).</li><li>Binnington, K. C., Kemp, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+tick+salivary+glands+in+feeding+and+disease+transmission.">Role of tick salivary glands in feeding and disease transmission.</a> Adv. Parasitol. 18, 315-339 (1980).</li><li>Guo, 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=Inhibition+of+neutrophil+function+by+two+tick+salivary+proteins.">Inhibition of neutrophil function by two tick salivary proteins.</a> Infect. Immun. 77, 2320-2329 (2009).</li><li>Montgomery, R. R., Lusitani, D., De Boisfleury Chevance, A., Malawista, S. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tick+saliva+reduces+adherence+and+area+of+human+neutrophils.">Tick saliva reduces adherence and area of human neutrophils.</a> Infect. Immun. 72, 2989-2994 (2004).</li><li>Lima, C. M., 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=Differential+infectivity+of+the+Lyme+disease+spirochete+Borrelia+burgdorferi+derived+from+Ixodes+scapularis+salivary+glands+and+midgut.">Differential infectivity of the Lyme disease spirochete Borrelia burgdorferi derived from Ixodes scapularis salivary glands and midgut.</a> J. Med. Entomol. 42, 506-510 (2005).</li><li>Severinova, 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=Co-inoculation+of+Borrelia+afzelii+with+tick+salivary+gland+extract+influences+distribution+of+immunocompetent+cells+in+the+skin+and+lymph+nodes+of+mice.">Co-inoculation of Borrelia afzelii with tick salivary gland extract influences distribution of immunocompetent cells in the skin and lymph nodes of mice.</a> Folia Microbiol. 50, 457-463 (2005).</li><li>Labuda, M., Jones, L. D., Williams, T., Nuttall, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enhancement+of+tick-borne+encephalitis+virus+transmission+by+tick+salivary+gland+extracts.">Enhancement of tick-borne encephalitis virus transmission by tick salivary gland extracts.</a> Med. Vet. Entomol. 7, 193-196 (1993).</li><li>Kariu, T., Coleman, A. S., Anderson, J. F., Pal, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methods+for+Rapid+Transfer+and+Localization+of+Lyme+Disease+Pathogens+Within+the+Tick+Gut.">Methods for Rapid Transfer and Localization of Lyme Disease Pathogens Within the Tick Gut.</a> J. Vis. Exp. (48), e2544 (2011).</li><li>Edwards, K. T., Goddard, J., Varela-Stokes, A. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Examination+of+the+internal+morphology+of+the+Ixodid+tick+Amblyomma+maculatum+koch,+(Acari:Ixodidae);+a+"How-to"+pictorial+dissection+guide.">Examination of the internal morphology of the Ixodid tick Amblyomma maculatum koch, (Acari:Ixodidae); a "How-to" pictorial dissection guide.</a> Midsouth Entomologist. 2, 28-39 (2009).</li><li>Ledin, K. E., 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=Borreliacidal+activity+of+saliva+of+the+tick+Amblyomma+americanum.">Borreliacidal activity of saliva of the tick Amblyomma americanum.</a> Med. Vet. Entomol. 19, 90-95 (2005).</li><li>Ribeiro, J. M., Zeidner, N. S., Ledin, K., Dolan, M. C., Mather, T. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=How+much+pilocarpine+contaminates+pilocarpine-induced+tick+saliva?.">How much pilocarpine contaminates pilocarpine-induced tick saliva?.</a> Med. Vet. Entomol. 18, 20-24 (2004).</li><li>Barker, R. W., Burris, E., Sauer, J. R., Hair, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Composition+of+tick+oral+secretions+obtained+by+three+different+collection+methods.">Composition of tick oral secretions obtained by three different collection methods.</a> J. Med. Entomol. 10, 198-201 (1973).</li><li>Burgdorfer, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hemolymph+test.+A+technique+for+detection+of+rickettsiae+in+ticks.">Hemolymph test. A technique for detection of rickettsiae in ticks.</a> Am. J. Trop. Med. Hyg. 19, 1010-1014 (1970).</li></ol>

No tenemos nada que revelar.

La colección de la hemolinfa de garrapatas, las glándulas salivales y la saliva es importante en el estudio de la transmitida por las garrapatas transmisión del patógeno, la prevalencia, la difusión, la proliferación y persistencia, tanto en la garrapata y el anfitrión 6,11-13,20,23,29. Hay varias formas para disecar una marca 30,31. Sin embargo, en la recogida de las glándulas salivales es fundamental para diseccionar la garrapata correctamente por lo que las glándulas salivales no se rom...

<table border="1"><tbody><tr><td> Reactivo </td><td> Empresa </td><td> Número de catálogo </td></tr><tr><td> Peróxido de hidrógeno </td><td> Pescador </td><td> H312-500 </td></tr><tr><td> Etanol </td><td> Acros </td><td> 61509-5000 </td></tr><tr><td> PBS </td><td> Boston Bioproductos </td><td> BM-2205 </td></tr><tr><td> Dumont Bellas fórceps (3C) </td><td> Pescador </td><td> NC9906085 </td></tr><tr><td> Tratada con silano portaobjetos de microscopio </td><td> Bioworld </td><td> 42763007-1 </td></tr><tr><td> Papanicolaou lápiz </td><td> Bioworld </td><td> 21750008-1 </td></tr><tr><td> Súper heladas, más portaobjetos de microscopio </td><td> Pescador </td><td> 12-550-18 </td></tr><tr><td> La pilocarpina </td><td> Sigma </td><td> P6503-5G </td></tr><tr><td> Cóctel de inhibidores de la proteasa </td><td> Sigma </td><td> P2714 </td></tr><tr><td> # 11 desechables bisturí </td><td> Pluma </td><td> 2975 # 11 </td></tr><tr><td> Plastilina no tóxica </td><td&gt; Fisher </td><td> S17307 </td></tr><tr><td> Los tubos capilares </td><td> Caza científica de vidrio, inc </td><td> 40A502 </td></tr></tbody></table>

saliva, salivary gland, and hemolymph collection from ixodes scapularis ticks

Las garrapatas se encuentran en todo el mundo y afectan a los seres humanos con muchas enfermedades transmitidas por garrapatas. Las garrapatas son vectores de patógenos que causan la enfermedad de Lyme transmitida por garrapatas y la fiebre recurrente (Borrelia spp.), Rocky Mountain la fiebre manchada (Rickettsia rickettsii), ehrlichiosis (Ehrlichia equi chaffeensis y E.), anaplasmosis (Anaplasma phagocytophilum), encefalitis (por garrapatas virus de la encefalitis transmitida), babesiosis (Babesia spp.), la fiebre de Colorado (Coltivirus), y la tularemia (Francisella tularensis) 1-8. Para ser transmitido de forma adecuada en el huésped estos agentes infecciosos regular la expresión génica diferencial, interactuar con las proteínas de garrapatas, y migran a través de la garrapata 3,9-13. Por ejemplo, el agente de la enfermedad de Lyme, Borrelia burgdorferi, se adapta a través de la expresión génica diferencial a las etapas de escasez y abundancia de ciclo enzoótico de la garrapata 14,15. Además, como una garrapata Ixodes consume unaharina de sangre de Borrelia replicarse y migrar del intestino medio en el hemocele, donde viajan a las glándulas salivales y se transmiten en el huésped con la saliva expulsada 9,16-19. Como una garrapata se alimenta del huésped por lo general responde con una fuerte respuesta inmune innata y hemostático 11,13,20-22. A pesar de estas respuestas de acogida, I. scapularis pueden alimentarse durante varios días debido a saliva de la garrapata contiene proteínas que son inmunomoduladores, agentes líticos, anticoagulantes y fibrinolysins para ayudar a alimentar la garrapata 3,11,20,21,23. Las actividades inmunomoduladoras que poseen saliva de la garrapata o extracto de las glándulas salivales (SGE) facilitan la transmisión, la proliferación y difusión de numerosos patógenos transmitidos por garrapatas 3,20,24-27. Para entender mejor cómo transmitida por garrapatas agentes infecciosos causan la enfermedad es esencial para diseccionar alimentando activamente garrapatas y recoger saliva de la garrapata. Este protocolo vídeo muestra la técnica de disección dela recogida de hemolinfa y la eliminación de las glándulas salivares de manera activa la alimentación I. scapularis ninfas después de 48 y 72 horas después de la colocación del ratón. También demuestran la toma de saliva de un adulto I. femenina scapularis garrapatas.

Watch this Scientific Journal Video about Saliva, Salivary Gland, and Hemolymph Collection from Ixodes scapularis Ticks at JoVE.com

Saliva, Salivary Gland, and Hemolymph Collection from Ixodes scapularis Ticks

La colección de la hemolinfa garrapata infectada, las glándulas salivales y la saliva es importante para estudiar la forma transmitida por garrapatas patógenos causan enfermedades. En este protocolo se demuestra cómo recoger las glándulas salivales y la hemolinfa de la alimentación<em&gt; Ixodes scapularis</em&gt; Ninfas. También demuestran la toma de saliva de la mujer<em&gt; I. scapularis</em&gt; Adultos.

La saliva, la glándula salival, y la Colección hemolinfa de las garrapatas Ixodes scapularis

Ticks are found worldwide and afflict humans with many tick-borne illnesses. Ticks are vectors for pathogens that cause Lyme disease and tick-borne ...

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Research

JoVE Journal

Immunology and Infection

Saliva, Salivary Gland, and Hemolymph Collection from Ixodes scapularis Ticks

29.2K vistas.  Centers for Disease Control and Prevention. La colección de la hemolinfa garrapata infectada, las glándulas salivales y la saliva es importante para estudiar la forma transmitida por garrapatas patógenos causan enfermedades. En este protocolo se demuestra cómo recoger las glándulas salivales y la hemolinfa de la alimentación Ixodes scapularis Ninfas. También demuestran la toma de saliva de la mujer I. scapularis Adultos.

Video: Saliva, Salivary Gland, and Hemolymph Collection from Ixodes scapularis Ticks

RNA Interference in Ticks

Ticks are obligate hematophagous ectoparasites of wild and domestic animals and humans, and are considered to be second worldwide to mosquitoes as vectors of human diseases1 and the most important vectors affecting cattle industry worldwide2. Ticks are classified in the subclass Acari, order Parasitiformes, suborder Ixodida and are distributed worldwide from Arctic to tropical regions3. Despite efforts to control tick infestations, these ectoparasites remain a serious problem for human and animal health4,5.
RNA interference (RNAi)6 is a nucleic acid-based reverse genetic approach that involves disruption of gene expression in order to determine gene function or its effect on a metabolic pathway. Small interfering RNAs (siRNAs) are the effector molecules of the RNAi pathway that is initiated by double-stranded RNA (dsRNA) and results in a potent sequence-specific degradation of cytoplasmic mRNAs containing the same sequence as the dsRNA trigger7-9. Post-transcriptional gene silencing mechanisms initiated by dsRNA have been discovered in all eukaryotes studied thus far, and RNAi has been rapidly developed in a variety of organisms as a tool for functional genomics studies and other applications10.
RNAi has become the most widely used gene-silencing technique in ticks and other organisms where alternative approaches for genetic manipulation are not available or are unreliable5,11. The genetic characterization of ticks has been limited until the recent application of RNAi12,13. In the short time that RNAi has been available, it has proved to be a valuable tool for studying tick gene function, the characterization of the tick-pathogen interface and the screening and characterization of tick protective antigens14. Herein, a method for RNAi through injection of dsRNA into unfed ticks is described. It is likely that the knowledge gained from this experimental approach will contribute markedly to the understanding of basic biological systems and the development of vaccines to control tick infestations and prevent transmission of tick-borne pathogens15-19.

A method for RNA interference (RNAi) by injection of dsRNA into unfed ticks is described. RNAi is the most widely used gene-silencing technique in ticks where the use of other methods of genetic manipulation has been limited.

La interferencia de ARN en las garrapatas

Ticks are obligate hematophagous ectoparasites of wild and domestic animals and humans, and are considered to be second worldwide to mosquitoes as vectors ...

Investigación

Inmunología e Infección

Hybridization in situ of Salivary Glands, Ovaries, and Embryos of Vector Mosquitoes

Mosquitoes are vectors for a diverse set of pathogens including arboviruses, protozoan parasites and nematodes. Investigation of transcripts and gene regulators that are expressed in tissues in which the mosquito host and pathogen interact, and in organs involved in reproduction are of great interest for strategies to reduce mosquito-borne disease transmission and disrupt egg development. A number of tools have been employed to study and validate the temporal and tissue-specific regulation of gene expression. Here, we describe protocols that have been developed to obtain spatial information, which enhances our understanding of where specific genes are expressed and their products accumulate. The protocol described has been used to validate expression and determine accumulation patterns of transcripts in tissues related to mosquito-borne pathogen transmission, such as female salivary glands, as well as subcellular compartments of ovaries and embryos, which relate to mosquito reproduction and development. 
 The following procedures represent an optimized methodology that improves the efficiency of various steps in the protocol without loss of target-specific hybridization signals. Guidelines for RNA probe preparation, dissection of soft tissues and the general procedure for fixation and hybridization are described in Part A, while steps specific for the collection, fixation, pre-hybridization and hybridization of mosquito embryos are detailed in Part B.

Hybridization in situ of Salivary Glands, Ovaries, and Embryos of Vector Mosquitoes

Temporal and spatial gene expression analyses have a crucial role in functional genomics. Whole-mount hybridization in situ is useful for determining the localization of transcripts within tissues and subcellular compartments. Here we outline a hybridization in situ protocol with modifications for specific target tissues in mosquitoes.

Hibridación<em&gt; In situ</em&gt; De las glándulas salivales, los ovarios y embriones de los mosquitos vectores

Mosquitoes are vectors for a diverse set of pathogens including arboviruses, protozoan parasites and nematodes.  Investigation of transcripts and gene ...

Feeding of Ticks on Animals for Transmission and Xenodiagnosis in Lyme Disease Research

Transmission of the etiologic agent of Lyme disease, Borrelia burgdorferi, occurs by the attachment and blood feeding of Ixodes species ticks on mammalian hosts. In nature, this zoonotic bacterial pathogen may use a variety of reservoir hosts, but the white-footed mouse (Peromyscus leucopus) is the primary reservoir for larval and nymphal ticks in North America. Humans are incidental hosts most frequently infected with B. burgdorferi by the bite of ticks in the nymphal stage. B. burgdorferi adapts to its hosts throughout the enzootic cycle, so the ability to explore the functions of these spirochetes and their effects on mammalian hosts requires the use of tick feeding. In addition, the technique of xenodiagnosis (using the natural vector for detection and recovery of an infectious agent) has been useful in studies of cryptic infection. In order to obtain nymphal ticks that harbor B. burgdorferi, ticks are fed live spirochetes in culture through capillary tubes. Two animal models, mice and nonhuman primates, are most commonly used for Lyme disease studies involving tick feeding. We demonstrate the methods by which these ticks can be fed upon, and recovered from animals for either infection or xenodiagnosis.

Lyme disease is the most commonly-reported vector-borne disease in North America. The causative agent, Borrelia burgdorferi is a spirochete bacterium transmitted by Ixodid ticks. Transmission and detection of infection in animal models is optimized by the use of tick feeding, which we describe here.

La alimentación de las garrapatas de los animales para Transmisión y xenodiagnóstico en Lyme Investigación de Enfermedades

Transmission of the etiologic agent of Lyme disease, Borrelia burgdorferi, occurs by the  attachment and blood feeding of Ixodes species ticks on ...

Rearing Ixodes scapularis, the Black-legged Tick: Feeding Immature Stages on Mice

Ixodes scapularis, the vector of Lyme disease, is one of the most important disease vectors in the eastern and Midwestern United States. This species is a three host tick that requires a blood meal from a vertebrate host for each development stage, and the adult females require a blood meal for reproduction. Larval ticks attach to their host for 3 - 5 days for feeding and drop off the host when fully engorged. This dependency on several different hosts and the lengthy attachment time for engorgement complicates tick rearing in the laboratory setting. However, to understand tick biology and tick-pathogen interactions, the production of healthy, laboratory-reared ticks is essential. Here, we demonstrate a simple, cost-effective protocol for immature tick feeding on mice. We modified the existing protocols for decreased stress on mice and increased tick feeding success and survival by using disposable cages without mesh bottoms to avoid contact of ticks with water contaminated with mice urine and feces.

Rearing Ixodes scapularis, the Black-legged Tick: Feeding Immature Stages on Mice

The production of healthy laboratory-reared ticks is essential to studies on tick biology, and tick-pathogen interactions. Here we demonstrate a simple protocol for immature tick feeding that is cost-effective and less stressful to mice.

La cría<em&gt; Ixodes scapularis,</em&gt; La garrapata de patas Negro: La alimentación de los estados inmaduros de ratones

Ixodes scapularis, the vector of Lyme disease, is one of the most important disease vectors in the eastern and Midwestern United States. This species is a ...

A Method for the Measurement of Salivary Gland Function in Mice

Patients with Sj&#246;gren&#39;s syndrome, an autoimmune disease affecting the exocrine glands, develop salivary gland inflammation and have reduced saliva production. Similarly, saliva production is severely compromised in patients receiving radiation treatment for head and neck cancers. Rodent models, developed to mimic these clinical conditions, facilitate an understanding of the disease pathogenesis and allow for the development of new therapeutic strategies. Therefore, the ability to accurately, reproducibly, and repeatedly measure salivary gland function in animal models is critical. Building on procedures previously described in the literature, a method was developed that meets these criteria and was used to evaluate salivary gland function in mice. An additional advantage of this new method is that it is easily mastered, and has little inter-operator variation. Salivary gland function is evaluated as the amount (weight or volume) or rate (mL/min) of saliva produced in response to pilocarpine stimulation. The collected saliva is a good source for the analyses of protein content, immunoglobulin concentrations, and other biomolecules.

Salivary gland hypofunction is a frequent consequence of autoimmune disease and radiation therapy. Reproducible evaluation of salivary gland function in mouse models of these diseases is a technical challenge. Here, a simple method for accurate and reproducible measurement of saliva production in mice is described.

Un método para la medición de la función de la glándula salival en ratones

Patients with Sj&#246;gren&#39;s syndrome, an autoimmune disease affecting the exocrine glands, develop salivary gland inflammation and have reduced ...

Preparation of Murine Submandibular Salivary Gland for Upright Intravital Microscopy

The submandibular salivary gland (SMG) is one of the three major salivary glands, and is of interest for many different fields of biological research, including cell biology, oncology, dentistry, and immunology. The SMG is an exocrine gland comprised of secretory epithelial cells, myofibroblasts, endothelial cells, nerves, and extracellular matrix. Dynamic cellular processes in the rat and mouse SMG have previously been imaged, mostly using inverted multi-photon microscope systems. Here, we describe a straightforward protocol for the surgical preparation and stabilization of the murine SMG in anesthetized mice for in vivo imaging with upright multi-photon microscope systems. We present representative intravital image sets of endogenous and adoptively transferred fluorescent cells, including the labeling of blood vessels or salivary ducts and second harmonic generation to visualize fibrillar collagen. In sum, our protocol allows for surgical preparation of mouse salivary glands in upright microscopy systems, which are commonly used for intravital imaging in the field of immunology.

We describe a protocol to surgically expose and stabilize the murine submandibular salivary gland for intravital imaging using upright intravital microscopy. This protocol is easily adaptable to other exocrine glands of the head and neck region of mice and other small rodents.

Preparación de glándula salival submaxilar murino para la microscopia Intravital vertical

The submandibular salivary gland (SMG) is one of the three major salivary glands, and is of interest for many different fields of biological research, ...

Isolation of Salivary Epithelial Cells from Human Salivary Glands for In Vitro Growth as Salispheres or Monolayers

The salivary glands are a site of significant interest for researchers interested in multiple aspects of human disease. One goal of researchers is to restore function of glands damaged by radiation therapies or due to pathologies associated with Sj&#246;gren's syndrome. A second goal of researchers is to define the mechanisms by which viruses replicate within glandular tissue where they can then gain access to salivary fluids important for horizontal transmission. These goals highlight the need for a robust and accessible in vitro salivary gland model that can be utilized by researchers interested in the above mentioned as well as related research areas. Here we discuss a simple protocol to isolate epithelial cells from human salivary glands and propagate them in vitro. Our protocol can be further optimized to meet the needs of individual studies. Briefly, salivary tissue is mechanically and enzymatically separated to isolate single cells or small clusters of cells. Selection for epithelial cells occurs by plating onto a basement membrane matrix in the presence of media optimized to promote epithelial cell growth. These resulting cultures can be maintained as three-dimensional clusters, termed "salispheres", or grown as a monolayer on treated plastic tissue culture dishes. This protocol results in the outgrowth of a heterogenous population of mainly epithelial cells that can be propagated for 5-8 passages (15-20 population doublings) before undergoing cellular senescence.

We present a method for isolating and cultivating primary human salivary gland-derived epithelial cells. These cells exhibit gene expression patterns consistent with them being of salivary epithelial origin and can be grown as salispheres on basement membrane matrices derived from Engelbreth-Holm-Swarm tumor cells or as monolayers on treated culture dishes.

Aislamiento de células epiteliales salivales de glándulas salivales humanas para el crecimiento in vitro como salisferas o monocapas

The salivary glands are a site of significant interest for researchers interested in multiple aspects of human disease. One goal of researchers is to ...

Purification of High Yield Extracellular Vesicle Preparations Away from Virus

One of the major hurdles in the field of extracellular vesicle (EV) research today is the ability to achieve purified EV preparations in a viral infection setting. The presented method is meant to isolate EVs away from virions (i.e., HIV-1), allowing for a higher efficiency and yield compared to conventional ultracentrifugation methods. Our protocol contains three steps: EV precipitation, density gradient separation, and particle capture. Downstream assays (i.e., Western blot, and PCR) can be run directly following particle capture. This method is advantageous over other isolation methods (i.e., ultracentrifugation) as it allows for the use of minimal starting volumes. Furthermore, it is more user friendly than alternative EV isolation methods requiring multiple ultracentrifugation steps. However, the presented method is limited in its scope of functional EV assays as it is difficult to elute intact EVs from our particles. Furthermore, this method is tailored towards a strictly research-based setting and would not be commercially viable.

This protocol isolates extracellular vesicles (EVs) away from virions with high efficiency and yield by incorporating EV precipitation, density gradient ultracentrifugation, and particle capture to allow for a streamlined workflow and a reduction of starting volume requirements, resulting in reproducible preparations for use in all EV research.

Purificación de preparaciones de vesícula extracelular de alto rendimiento lejos del virus

One of the major hurdles in the field of extracellular vesicle (EV) research today is the ability to achieve purified EV preparations in a viral infection ...

Disclosing Hemolymph Collection and Inoculation of Metarhizium Blastospores into Rhipicephalus Microplus Ticks Towards Invertebrate Pathology Studies

Ticks are obligate hematophagous ectoparasites and Rhipicephalus microplus has great importance in veterinary medicine because it causes anemia, weight loss, depreciation of the animals&#39; leather and also can act as a vector of several pathogens. Due to the exorbitant costs to control these parasites, damage to the environment caused by the inappropriate use of chemical acaricides, and the increased resistance against traditional parasiticides, alternative control of ticks, by the use of entomopathogenic fungi, for example, has been considered an interesting approach. Nevertheless, few studies have demonstrated how the tick&#39;s immune system acts to fight these entomopathogens. Therefore, this protocol demonstrates two methods used for entomopathogen inoculation into engorged females and two techniques used for hemolymph collection and hemocytes harvesting. Inoculation of pathogens at the leg insertion in the tick female&#39;s body allows evaluation of females biologic parameters unlike the inoculation between the scutum and capitulum, which frequently damages Gen&#233;&#39;s organ. Dorsal hemolymph collection yielded a higher volume recovery than collection through the legs. Some limitations of tick hemolymph collection and processing include i) high rates of hemocytes&#39; disruption, ii) hemolymph contamination with disrupted midgut, and iii) low hemolymph volume recovery. When hemolymph is collected through the leg cutting, the hemolymph takes time to accumulate at the leg opening, favoring the clotting process. In addition, fewer hemocytes are obtained in the collection through the leg compared to the dorsal collection, even though the first method is considered easier to be performed. Understanding the immune response in ticks mediated by entomopathogenic agents helps to unveil their pathogenesis and develop new targets for tick control. The inoculation processes described here require very low technological resources and can be used not only to expose ticks to pathogenic microorganisms. Similarly, the collection of tick hemolymph may represent the first step for many physiological studies.

Disclosing Hemolymph Collection and Inoculation of Metarhizium Blastospores into Rhipicephalus Microplus Ticks Towards Invertebrate Pathology Studies

Analysis of tick hemolymph represents an important source of information on how some pathogens cause disease and how ticks immunologically respond to this infection. The present study demonstrates how to inoculate fungal propagules and collect hemolymph from Rhipicephalus microplus engorged females.

Divulgar la colección hemolymph y la inoculación de Metarhizium blastosporos en las garrapatas de rhipicephalus microplus hacia los estudios de patología de invertebrados

Ticks are obligate hematophagous ectoparasites and Rhipicephalus microplus has great importance in veterinary medicine because it causes anemia, weight ...

Imágenes en vivo en tiempo real de la regeneración de la glándula submandibular del ratón

Interrogating Cell-Cell Interactions in the Salivary Gland via Ex Vivo Live Cell Imaging

Salivary gland regeneration is a complex process involving intricate interactions among various cell types. Recent studies have shed light on the pivotal role played by macrophages in driving the regenerative response. However, our understanding of this critical role has primarily relied on static views obtained from fixed tissue biopsies. To overcome this limitation and gain insights into these interactions in real time, this study outlines a comprehensive protocol for culturing salivary gland tissue ex vivo and capturing live images of cell migration.
The protocol involves several key steps: First, mouse submandibular salivary gland tissue is carefully sliced using a vibratome and then cultured at an air-liquid interface. These slices can be intentionally injured, for instance, through exposure to radiation, to induce cellular damage and trigger the regenerative response. To track specific cells of interest, they can be endogenously labeled, such as by utilizing salivary gland tissue collected from genetically modified mice where a particular protein is marked with green fluorescent protein (GFP). Alternatively, fluorescently-conjugated antibodies can be employed to stain cells expressing specific cell surface markers of interest. Once prepared, the salivary gland slices are subjected to live imaging using a high-content confocal imaging system over a duration of 12 h, with images captured at 15 min intervals. The resulting images are then compiled to create a movie, which can subsequently be analyzed to extract valuable cell behavior parameters. This innovative method provides researchers with a powerful tool to investigate and better understand macrophage interactions within the salivary gland following injury, thereby advancing our knowledge of the regenerative processes at play in this dynamic biological context.

Autor destacado: Estudio de las interacciones entre macrófagos y células epiteliales en la regeneración de glándulas salivales después de una lesión 

Immunofluorescent imaging is constrained by the ability to observe complex, time-dependent biological processes in just a single snapshot in time. This study outlines a live-imaging approach conducted on precision-cut mouse submandibular gland slices. This approach allows for the real-time observation of cell-cell interactions during homeostasis and the processes of regeneration and repair.

Interrogación de las interacciones célula-célula en la glándula salival a través de imágenes de células vivas ex vivo

Salivary gland regeneration is a complex process involving intricate interactions among various cell types. Recent studies have shed light on the pivotal ...