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<ol><li>Scheele, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Multiphoton+intravital+microscopy+of+rodents%5BTitle%5D)+AND+%22Nat+Rev+Methods+Primers%22%5BJournal%5D)">Multiphoton intravital microscopy of rodents.</a> Nat Rev Methods Primers. 2 (1), 1-26 (2022).</li>
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<li>Nooshabadi, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Intravital+excitation+increases+detection+sensitivity+for+pulmonary+tuberculosis+by+whole-body+imaging+with+beta-lactamase+reporter+enzyme+fluorescence%5BTitle%5D)+AND+%22J+Biophotonics%22%5BJournal%5D)">Intravital excitation increases detection sensitivity for pulmonary tuberculosis by whole-body imaging with beta-lactamase reporter enzyme fluorescence.</a> J Biophotonics. 10 (6-7), 821-829 (2017).</li>
<li>Sharan, R., Yang, H. J., Sule, P., Cirillo, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Imaging+Mycobacterium+tuberculosis+in+Mice+with+Reporter+Enzyme+Fluorescence%5BTitle%5D)+AND+%22J+Vis+Exp%22%5BJournal%5D)">Imaging Mycobacterium tuberculosis in Mice with Reporter Enzyme Fluorescence.</a> J Vis Exp. (132), e56801(2018).</li>
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<li>Barlerin, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Biosafety+Level+3+setup+for+multiphoton+microscopy+in+vivo%5BTitle%5D)+AND+%22Sci+Rep%22%5BJournal%5D)">Biosafety Level 3 setup for multiphoton microscopy in vivo.</a> Sci Rep. 7 (1), 571(2017).</li>
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<li>Ovchinnikov, D. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Expression+of+Gal4-dependent+transgenes+in+cells+of+the+mononuclear+phagocyte+system+labeled+with+enhanced+cyan+fluorescent+protein+using+Csf1r-Gal4VP16%2FUAS-ECFP+double-transgenic+mice%5BTitle%5D)+AND+%22J+Leukoc+Biol%22%5BJournal%5D)">Expression of Gal4-dependent transgenes in cells of the mononuclear phagocyte system labeled with enhanced cyan fluorescent protein using Csf1r-Gal4VP16/UAS-ECFP double-transgenic mice.</a> J Leukoc Biol. 83 (2), 430-433 (2008).</li>
<li>Cohen, S. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Alveolar+Macrophages+Provide+an+Early+Mycobacterium+tuberculosis+Niche+and+Initiate+Dissemination%5BTitle%5D)+AND+%22Cell+Host+Microbe%22%5BJournal%5D)">Alveolar Macrophages Provide an Early Mycobacterium tuberculosis Niche and Initiate Dissemination.</a> Cell Host Microbe. 24 (3), 439-446.e4 (2018).</li>
<li>Pandey, A. K., Sassetti, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mycobacterial+persistence+requires+the+utilization+of+host+cholesterol%5BTitle%5D)+AND+%22Proc+Natl+Acad+Sci+U+S+A%22%5BJournal%5D)">Mycobacterial persistence requires the utilization of host cholesterol.</a> Proc Natl Acad Sci U S A. 105 (11), 4376-4380 (2008).</li>
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</ol>

<table><tbody><tr><td>15 ml conical tube</td><td>Corning</td><td>430790</td><td></td></tr><tr><td>2-0 silk tie&nbsp;</td><td>Ethicon, Inc</td><td>LA55G</td><td></td></tr><tr><td>5 mm coverslip</td><td>Electron Microscopy Sciences</td><td>72296-05</td><td></td></tr><tr><td>5 mm disposable biopsy punch</td><td>Integra</td><td>33-35-SH</td><td></td></tr><tr><td>5-0 silk purse-string suture&nbsp;</td><td>Ethicon, Inc</td><td>640G</td><td></td></tr><tr><td>Blunt micro-dissecting scissors</td><td>Roboz</td><td>RS-5980</td><td></td></tr><tr><td>Buprenorphine</td><td>Hospira</td><td>0409-2012-32</td><td>Injeção de 10 &mu;L de buprenorfina (solução de estoque de 0,3 mg/mL) diluída em 90 &mu;L de PBS estéril (dose final = 0,1 mg/kg)&nbsp;</td></tr><tr><td>Compressed air canister</td><td>Falcon</td><td>DPSJB-12</td><td></td></tr><tr><td>Cyanoacrylate adhesive</td><td>Loctite</td><td>234790</td><td></td></tr><tr><td>D-calcium pantothenate</td><td>Acros</td><td>243305000</td><td></td></tr><tr><td>ECM 630 Electroporation System Complete, with Safety stand</td><td>BTX Harvard Apparatus</td><td>45-0051</td><td></td></tr><tr><td>Electroporation cuvette</td><td>Fisher Scientific</td><td>FB102</td><td></td></tr><tr><td>FITC-Dextran</td><td>Sigma-Aldrich</td><td>FD150S-1G</td><td>150 kDa</td></tr><tr><td>Glycerol</td><td>Fisher Chemicals</td><td>G33-1</td><td></td></tr><tr><td>Graefe forceps</td><td>Roboz</td><td>RS-5135</td><td>Para agarrar e elevar a costela durante o procedimento de implantação da janela pulmonar</td></tr><tr><td>Isoflurane</td><td>&nbsp;Pivetal</td><td>78949580</td><td></td></tr><tr><td>Isoflurane vaporizer</td><td>SurgiVet</td><td>VCT302</td><td></td></tr><tr><td>Kanamycin</td><td>Goldbio.com</td><td>K-120-25</td><td></td></tr><tr><td>L-arginine monohydrochloride</td><td>Sigma-Aldrich</td><td>A5131-100G</td><td></td></tr><tr><td>L-leucine</td><td>Sigma-Aldrich</td><td>L8000</td><td></td></tr><tr><td>MacBlue mice</td><td>Jackson Labs</td><td>Strain #: 026051</td><td></td></tr><tr><td>Middlebrook 7H10 Agar</td><td>Difco</td><td>262710</td><td></td></tr><tr><td>Middlebrook 7H9 broth</td><td>Difco</td><td>271310</td><td></td></tr><tr><td>Middlebrook OADC Enrichment</td><td>Becton Dickinson&nbsp;</td><td>212351</td><td></td></tr><tr><td>Murine ventilator</td><td>Kent Scientific</td><td>PS-02</td><td>PhysioSuite</td></tr><tr><td>Nair</td><td>Church &amp; Dwight Co., Inc.</td><td>40002957</td><td></td></tr><tr><td>PBS</td><td>Corning</td><td>21-031-CV</td><td></td></tr><tr><td>PETG media inkwell bottle</td><td>Thermo Scientific</td><td>342020</td><td></td></tr><tr><td>PhysioSuite</td><td>Kent Scientific</td><td>PS-5623</td><td>Para monitorar o volume tidal do mouse&rsquo;s via intubação durante a cirurgia</td></tr><tr><td>Plasmid pYUB1169</td><td>Plasmid custom made</td><td></td><td></td></tr><tr><td>Rag2 KO mice</td><td>Jackson Labs</td><td>Strain #: 008449</td><td></td></tr><tr><td>Small animal lung inflation bulb</td><td>Harvard Apparatus</td><td>72-9083</td><td></td></tr><tr><td>Sodium propionate</td><td>Thermo Scientific</td><td>A17440.36</td><td></td></tr><tr><td>Sonicator</td><td>Fisher Scientific</td><td>FB705</td><td></td></tr><tr><td>Sorval Centrifuge X pro series</td><td>Thermo Scientific</td><td>75009020</td><td></td></tr><tr><td>Spectrophotometer Genesys 10 UV</td><td>Thermo Spectronic</td><td>335903</td><td></td></tr><tr><td>SurgiSuite</td><td>Kent Scientific</td><td>SURGI-M02</td><td>Multi-Functional Surgical Platform for Mice, with Warming</td></tr><tr><td>Tracheal catheter</td><td>Exelint International</td><td>26746</td><td>22G</td></tr><tr><td>Tyloxapol</td><td>Sigma-Aldrich</td><td>T0307-50G</td><td></td></tr><tr><td>Vacuum pickup system metal probe&nbsp;</td><td>Ted Pella, Inc.</td><td>528-112</td><td></td></tr></tbody></table>

imagem intravital de mycobacterium tuberculosis auxotrófico atenuado no pulmão murino durante infecção intravascular precoce

Intravital Imaging of Attenuated Auxotrophic Mycobacterium tuberculosis in Murine Lung during Early Intravascular Infection

Imagem intravital de Mycobacterium tuberculosis auxotrófico atenuado no pulmão murino durante infecção intravascular precoce

Intravital microscopy enables direct visualization of dynamic cellular processes within intact tissues. However, its application to Mycobacterium ...

intravital-imaging-attenuated-auxotrophic-mycobacterium-tuberculosis

Research

JoVE Journal

Biology

Biologia

Fonte: Niu, L., et al. Visualização da morte celular lítica de macrófagos durante infecção por micobactérias em embriões de peixe-zebra via microscopia intravital. J. Vis. Exp. (2019)Este vídeo demonstra o uso da microscopia confocal intravital para visualizar a morte celular lítica de macrófagos em embriões de peixe-zebra após infecção por Mycobacterium marinum .

Imagens ao vivo da morte celular de macrófagos durante infecção por micobactérias em embriões de peixe-zebra

Fonte: Bernut, A., et al. Decifração e Imagem da Patogênese e Cordamento do Mycobacterium abscessus em embriões de peixe-zebra. J. Vis. Exp. (2015)Este vídeo demonstra um protocolo que utiliza embriões transgênicos de peixe-zebra expressando macrófagos fluorescentes e infectados com a cepa áspera fluorescente Mycobacterium abscessus para visualizar as interações hospedeiro-patógeno. Ele revela como o cordão extracelular possibilita a evasão imunológica, promovendo a sobrevivência bacteriana e...

Imagens ao vivo das interações hospedeiro-patógeno em embriões de peixe-zebra infectados com Mycobacterium abscessus

O citomegalovírus (HCMV) A infecção humana de recém-nascidos representa uma importante causa de retardo mental, mas os eventos moleculares que levam à induzida por vírus patogênese ainda são pouco compreendidos. Para investigar a dinâmica da infecção cerebral, adaptamos...

Utilização de<em&gt; In vivo</em&gt; Imagem para monitorar a progressão da infecção pelo citomegalovírus Experimental Rato em recém-nascidos

Tuberculose e malária são duas das infecções mais prevalentes em seres humanos e as principais causas de morbidade e mortalidade em populações empobrecidas nos trópicos. Nós estabelecemos um sistema de modelo experimental para estudar resultado de malária tuberculose co-infecção em camundongos após o desafio com ambos os patógenos através de sua rota natural de infecção.

Um modelo experimental para estudar a Tuberculose-Malária Coinfection sobre Transmissão Natural de Mycobacterium tuberculosis E Plasmodium berghei</i

Streptococcus pneumoniae é o patógeno principal causador da pneumonia adquirida na comunidade grave e responsável por mais de 2 milhões de mortes em todo o mundo. O impacto dos fatores bacterianos implicados em fitness ou virulência pode ser monitorado em tempo real em uma pneumonia ou bacteremia modelo aguda mouse usando bactérias bioluminescentes.

A seguir em tempo real o impacto de fatores de virulência do pneumococo em um mouse Pneumonia aguda Modelo Usando bactérias bioluminescentes

Embriões de peixe-zebra opticamente transparentes são amplamente utilizados para estudar e visualizar em tempo real as interações entre microrganismos patogênicos e as células imunes inatas. A microinjeção de Mycobacterium abscessus, combinada com imagens de fluorescência, é usada para examinar características patogênicas essenciais, como a formação de cordão em embriões de peixe-zebra.

Decifrar e Imagem Patogênese e Cording de<em&gt; Mycobacterium abscessus</em&gt; Em embriões Zebrafish

Descrevemos a imagem óptica de camundongos infectados com Mycobacterium tuberculosis (M. tuberculosis) usando fluorescência de enzima repórter (REF). Esse protocolo proporciona uma detecção sensível e específica de M. tuberculosis em modelos animais pré-clínicos para investigação patogenia, terapêutica e vacina.

Mycobacterium tuberculosis em camundongos com fluorescência de enzima do repórter de imagem

Neste estudo, apresentamos um protocolo para executar dois fotões intravital imagem e célula interação análise na mucosa traqueal murino após infecção com vírus da gripe. O presente protocolo será relevante para pesquisadores estudando a dinâmica de células imunes durante infecções respiratórias.

Interação de célula de imagem latente na Mucosa traqueal durante a infecção de vírus de Influenza usando microscopia Intravital dois fotões

Este protocolo descreve uma técnica para visualizar o comportamento do macrófago e a morte no zebrafish embrionário durante a infecção do marinum de Mycobacterium. Etapas para a preparação de bactérias, infecção dos embriões e microscopia intravital estão incluídas. Esta técnica pode ser aplicada à observação do comportamento celular e da morte em cenários semelhantes envolvendo infecção ou inflamação estéril.

Visualização da morte de células ticas de macrófago durante infecção micobacteriana em embriões de zebrafish via microscopia intravital

Aqui, apresentamos um protocolo para obter imagens intravitais do pulmão esquerdo de camundongo transplantado usando microscopia de dois fótons. Isso representa uma ferramenta valiosa para estudar a dinâmica e as interações celulares em tempo real após o transplante de pulmão murino.

Microscopia intravital de dois fótons do pulmão de camundongo transplantado

Aqui apresentamos um protocolo para realizar uma inoculação intradérmica de baixa dose de Mycobacterium avium em uma orelha de camundongo para refletir condições semelhantes à exposição.

Inoculação intradérmica de Mycobacterium avium na orelha de camundongo

Albert Einstein College of Medicine.  Aqui, apresentamos um protocolo para imagem intravital de um Mycobacterium tuberculosis aprovado por BSL-2 atenuado. Cepa no pulmão murino usando uma janela pulmonar permanente. Essa abordagem permite a análise longitudinal com resolução de célula única das interações precoces hospedeiro-patógeno, incluindo localização bacilar, dinâmica vascular e captação de macrófagos dentro do mesmo microambiente pulmonar.

Imagem intravital de Mycobacterium tuberculosis auxotrófico atenuado no pulmão murino durante infecção intravascular precoce

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