<ol>
	<li>Baker, D. G., Fox, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chapter+23.">Chapter 23.</a> The Mouse in Biomedical Research: Diseases. Diseases Vol. 2, (2007).</li><li>Baker, D. G., Baker, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chapter+11.">Chapter 11.</a> Flynn's Parasites of Laboratory Animals. , 303-398 (2007).</li><li>Owen, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Parasites of Laboratory Animals. 12, (1992).</li><li>Pritchett, K. R., Fox, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chapter+22.">Chapter 22.</a> The Mouse in Biomedical Research: Diseases. Diseases Vol. 2, (2007).</li><li>Smith, P. H., Wiles, S. E., Malone, J. B., Monahan, C. M., Baker, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chapter+1.">Chapter 1.</a> Flynn's Parasites of Laboratory Animals. , 1-13 (2007).</li><li>Wasson, K., Fox, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chapter+21.">Chapter 21.</a> The Mouse in Biomedical Research: Diseases. Vol. 2, 517-550 (2007).</li></ol>

המחברים הם כל העובדים של צ&#39;ארלס ריבר, ספקית של בדיקות אבחון ריאגנטים, כולל הבדיקות שתוארו לעיל.

כאשר עובד במעבדה, בטיחות צריכה להיות תמיד דאגה. זכור ללבוש ציוד מגן מתאים כאשר עובדים עם בעלי חיים כדי לנקות תחנת העבודה שלך עם חומר חיטוי לפני ואחרי. שיטות אלה נועדו בעיקר כדי למצוא טפילים של מכרסמים במעבדה במקומות שנבדקו, כלומר, הם יכולים לזהות אקזוטיים או טפילים נדירים ביותר, כמו גם את ותולעי סיכה יותר נפוצים קרדית פרווה. למרות שהם חלים גם על מינים אחרים, מכרסמים בר ייתכן טפילים נוספים במקומות כגון subcutis הכבד, המוח, לא הוערכו על ידי השיטות הנ&quot;ל.

<table border="1" style=";text-align:right;direction:rtl"><tbody><tr><td> שם מגיב </td><td> חברה </td><td> מספר קטלוגי </td><td> תגובות </td></tr><tr><td> קרש חיתוך </td><td> Thermo Electron Corp </td><td> חתול # 36114 </td><td></td></tr><tr><td> מספריים קטנים </td><td> ROBOZ </td><td> RS-5910, G204 </td><td> להבי 23mm, 3.5 &quot;אורך, ישר </td></tr><tr><td> בינוני מספריים </td><td> ROBOZ </td><td> RS-6808, G207 </td><td> 5 &quot; </td></tr><tr><td> מלקחיים-מעוקל </td><td> ROBOZ </td><td> RS-8254 (M1/21004) </td><td> 4.5 &quot;, משונן, עקומה קלה </td></tr><tr><td> מלקחיים-Microdissecting </td><td> ROBOZ </td><td> RS-5238 </td><td> הדסון-(אוולד) </td></tr><tr><td> מלקחיים, רקמות מלקחיים </td><td> ROBOZ </td><td> RS-8160 </td><td> עכברוש שן </td></tr><tr><td> מתכת בדיקה </td><td> VWR </td><td> חתול # 25778-000 </td><td></td></tr><tr><td> Hemostats </td><td> יתרון </td><td> V97-48 </td><td></td></tr><tr><td> המוליך מקלות </td><td> הפוריטנים </td><td> Applicators 6in, שופט # 807 </td><td></td></tr><tr><td> הביתור מיקרוסקופ </td><td> אולימפוס </td><td> SZ51, שוט, ACE1 </td><td></td></tr><tr><td> צלחת פטרי </td><td> VWR </td><td> חתול 100mm, # 3401PDNL </td><td></td></tr><tr><td> כיסוי תלושי </td><td> VWR </td><td> מיקרו זכוכית לכסות, חתול # 48366-067 </td><td></td></tr><tr><td> שקופיות </td><td> VWR </td><td> VistaVision שקופיות מיקרוסקופ חתול # 16004-368 </td><td></td></tr><tr><td> Inoculating לולאה </td><td> VWR </td><td> חתול # 50815-040 </td><td></td></tr><tr><td> מיקרוסקופ אור </td><td> אולימפוס </td><td> דגם # BX41TF </td><td></td></tr><tr><td> מעבדה בתנור </td><td> קווינסי מעבדה בע&quot;מ </td><td> 10 דגם מעבדה תנור </td><td></td></tr><tr><td> סרכזת </td><td> Beckman Coulter </td><td> אלגרה X-12R </td><td></td></tr><tr><td> Vortexer </td><td> VWR </td><td> ה-Mini-Vortexer </td><td></td></tr><tr><td> מבחנה </td><td> קימבל-Chase </td><td> צינור 15 מ&quot;ל צנטריפוגות חד פעמיות, חתול # 73790-15 </td><td></td></tr><tr><td> כיסוי תלושי </td><td> VWR </td><td> פלסטיק לכיסוי מיקרוסקופ מחליק, חתול 22mm, # 48376-049 </td><td></td></tr><tr><td> כובעי הלבן </td><td> VWR </td><td> חתול # 60869-089 </td><td></td></tr><tr><td> יוד </td><td> רולי ביוכימיים המכון </td><td> חתול # SO-364 </td><td></td></tr><tr><td> אבץ סולפט </td><td> סיגמא אולדריץ </td><td> חתול # 1000917519, Z4750-500 גרם </td><td></td></tr><tr><td> סוכרוז </td><td> Mallinckrodt כימיקלים </td><td> חתול # 8360-06 </td><td></td></tr><tr><td> פנול </td><td> EMD </td><td> חתול # PX0510-1 </td><td></td></tr><tr><td> צלופן סרט </td><td> סטייפלס </td><td> קלטת חתול הנעלמה, # 504712 </td><td></td></tr><tr><td> שמן מינרלי </td><td> Mallickrodt </td><td> חתול # 6358 </td><td></td></tr></tbody></table>

diagnosis of ecto- and endoparasites in laboratory rats and mice

טפילים פנימיים וחיצוניים להישאר דאגה משמעותית מתקני מכרסם מעבדה, מתקני מחקר רבים הנמל כמה חיות parasitized. לפני היציאה בדיקה של בעלי חיים טפילים, שני דברים צריך להיחשב. אחת: מה שימוש ייעשה במידע שנאסף, ושני: אשר הבדיקה המתאימה ביותר. בידיעה בעלי חיים parasitized יכול להיות משהו במתקן מקבל, אבל לעיתים קרובות יש צורך לטפל בבעלי חיים, ואז לקבוע את יעילות הטיפול. טפילים ניתן לזהות אצל בעלי חיים באמצעות טכניקות שונות, כולל דגימות שנלקחו מבעלי חיים או מורדמים. מבחינה היסטורית, עם רגישות בדיקות האבחון הגדולים הנדרשים המתת חסד של בעלי חיים, למרות PCR אפשרה רגישות גבוהה בדיקות עבור מספר סוגים של טפיל. מאמר זה מדגים את הליכי זיהוי של אנדו ו ectoparasites בעכברים וחולדות. ההליכים באותו החלות על מכרסמים אחרים, אם כי מינים של טפילים מצאו יהיה שונה.

Watch this Scientific Journal Video about Diagnosis of Ecto- and Endoparasites in Laboratory Rats and Mice at JoVE.com

Diagnosis of Ecto- and Endoparasites in Laboratory Rats and Mice

מאמר זה מתאר את ההליכים השונים חולדות ועכברים ההקרנה לזהות אנדו או ectoparasitism. מבחני אבחון כמה יהיה הפגינו, הן אלה מתאימים לשימוש בבעלי חיים לבין אלה המשמשים לאחר המתת חסד של החיה. צילומים לסייע בזיהוי של טפילים חולדה ועכבר ייכלל.

אבחון חיצוני ו Endoparasites בחולדות מעבדה ועכברים

Internal and external parasites remain a significant concern in laboratory rodent facilities, and many research facilities harbor some parasitized ...

diagnosis-of-ecto-and-endoparasites-in-laboratory-rats-and-mice

Research

JoVE Journal

Immunology and Infection

41.0K Views.  Charles River. מאמר זה מתאר את ההליכים השונים חולדות ועכברים ההקרנה לזהות אנדו או ectoparasitism. מבחני אבחון כמה יהיה הפגינו, הן אלה מתאימים לשימוש בבעלי חיים לבין אלה המשמשים לאחר המתת חסד של החיה. צילומים לסייע בזיהוי של טפילים חולדה ועכבר ייכלל.

Video: Diagnosis of Ecto- and Endoparasites in Laboratory Rats and Mice

Orthotopic Hind-Limb Transplantation in Rats

Composite tissue allotransplantation (CTA) now represents a valid therapeutic option after the loss of a hand, forearm or digits and has become a novel therapeutic entity in reconstructive surgery. However, long term high-dose multi-drug immunosuppressive therapy is required to ensure graft survival, bearing the risk of serious side effects which halters broader application. Further progression in this field may depend on better understanding of basic immunology and ischemia reperfusion injury in composite tissue grafts.
To date, orthotopic hind limb transplantation in rats has been the preferred rodent model for reconstructive transplantation (RT), however, it is an extremely demanding procedure that requires extraordinary microsurgical skills for reattachment of vasculature, bones, muscles and nerves.
We have introduced the vascular cuff anastomosis technique to this model, providing a rapid and reliable approach to rat hind limb transplantation. This technique simplifies and shortens the surgical procedure and enables surgeons with basic microsurgical experience to successfully perform the operation with high survival and low complication rates. The technique seems to be well suited for immunological as well as ischemia reperfusion injury (IRI) studies.

Here we describe the orthotopic rat hind-limb transplantation procedure, which seems to be the gold standard in vivo model for composite tissue allotransplantation research.

השתלת orthotopic לימב הינד-בחולדות

Composite tissue allotransplantation (CTA) now represents a valid therapeutic option after the loss of a hand, forearm or digits and has become a novel ...

Microwave Assisted Rapid Diagnosis of Plant Virus Diseases by Transmission Electron Microscopy

Investigations of ultrastructural changes induced by viruses are often necessary to clearly identify viral diseases in plants. With conventional sample preparation for transmission electron microscopy (TEM) such investigations can take several days1,2 and are therefore not suited for a rapid diagnosis of plant virus diseases. Microwave fixation can be used to drastically reduce sample preparation time for TEM investigations with similar ultrastructural results as observed after conventionally sample preparation3-5. Many different custom made microwave devices are currently available which can be used for the successful fixation and embedding of biological samples for TEM investigations5-8. In this study we demonstrate on Tobacco Mosaic Virus (TMV) infected Nicotiana tabacum plants that it is possible to diagnose ultrastructural alterations in leaves in about half a day by using microwave assisted sample preparation for TEM. We have chosen to perform this study with a commercially available microwave device as it performs sample preparation almost fully automatically5 in contrast to the other available devices where many steps still have to be performed manually6-8 and are therefore more time and labor consuming. As sample preparation is performed fully automatically negative staining of viral particles in the sap of the remaining TMV-infected leaves and the following examination of ultrastructure and size can be performed during fixation and embedding.

This study describes a method that allows the rapid and clear diagnosis of plant virus diseases in about half a day by using a combination of microwave assisted plant sample preparation for transmission electron microscopy and negative staining methods.

מיקרוגל Assisted אבחון מהיר של מחל וירוס צמח על ידי מיקרוסקופי אלקטרוני הילוכים

Investigations of ultrastructural changes induced by viruses are often necessary to clearly identify viral diseases in plants. With conventional sample ...

Basophil Activation Test for Investigation of IgE-Mediated Mechanisms in Drug Hypersensitivity

Hypersensitivity reactions against non-steroidal anti-inflammatory drugs (NSAIDs) like propyphenazone (PP) and diclofenac (DF) can manifest as Type I-like allergic reactions 1. In clinical practice, diagnosis of drug hypersensitivity is mainly performed by patient history, as skin testing is not reliable and oral provocation testing bears life-threatening risks for the patient 2. Hence, evidence for an underlying IgE-mediated pathomechanism is hard to obtain. 
Here, we present an in vitro method based on the use of human basophils derived from drug-hypersensitive patients that mimics the allergic effector reaction in vivo. As basophils of drug-allergic patients carry IgE molecules specific for the culprit drug, they become activated upon IgE receptor crosslinking and release allergic effector molecules. The activation of basophils can be monitored by the determination of the upregulation of CD63 surface expression using flow cytometry 3. 
In the case of low molecular weight drugs, conjugates are designed to enable IgE receptor crosslinking on basophils. As depicted in Figure 1, two representatives of NSAIDs, PP and DF, are covalently bound to human serum albumin (HSA) via a carboxyl group reacting with the primary amino group of lysine residues. DF carries an intrinsic carboxyl group and, thus, can be used directly 4, whereas a carboxyl group-containing derivative of PP had to be organochemically synthesized prior to the study 1.
The coupling degree of the low molecular weight compounds on the protein carrier molecule and their spatial distribution is important to guarantee crosslinking of two IgE receptor molecules. The here described protocol applies high performance-size exclusion chromatography (HPSEC) equipped with a sequential refractive index (RI) and ultra violet (UV) detection system for determination of the coupling degree.
As the described methodology may be applied for other drugs, the basophil activation test (BAT) bears the potential to be used for the determination of IgE-mediated mechanisms in drug hypersensitivity. Here, we determine PP hypersensitivity as IgE-mediated and DF hypersensitivity as non-IgE-mediated by BAT.

Basophil activation test is a potent tool for the detection of IgE-dependent allergies in vitro. Here, an optimized protocol for basophil activation test is used to investigate drug hypersensitivity. A method for the efficient production of covalent drug-protein conjugates and their physicochemical characterization is described.

Basophil מבחן הפעלה לחקירות IgE בתיווך מנגנונים של תרופות רגישות יתר

Hypersensitivity reactions against non-steroidal anti-inflammatory drugs (NSAIDs) like propyphenazone (PP) and diclofenac (DF) can manifest as Type I-like ...

Safety Precautions and Operating Procedures in an (A)BSL-4 Laboratory: 1. Biosafety Level 4 Suit Laboratory Suite Entry and Exit Procedures

Biosafety level 4 (BSL-4) suit laboratories are specifically designed to study high-consequence pathogens for which neither infection prophylaxes nor treatment options exist. The hallmarks of these laboratories are: custom-designed airtight doors, dedicated supply and exhaust airflow systems, a negative-pressure environment, and mandatory use of positive-pressure (&#8220;space&#8221;) suits. The risk for laboratory specialists working with highly pathogenic agents is minimized through rigorous training and adherence to stringent safety protocols and standard operating procedures. Researchers perform the majority of their work in BSL-2 laboratories and switch to BSL-4 suit laboratories when work with a high-consequence pathogen is required. Collaborators and scientists considering BSL-4 projects should be aware of the challenges associated with BSL-4 research both in terms of experimental technical limitations in BSL-4 laboratory space and the increased duration of such experiments. Tasks such as entering and exiting the BSL-4 suit laboratories are considerably more complex and time-consuming compared to BSL-2 and BSL-3 laboratories. The focus of this particular article is to address basic biosafety concerns and describe the entrance and exit procedures for the BSL-4 laboratory at the NIH/NIAID Integrated Research Facility at Fort Detrick. Such procedures include checking external systems that support the BSL-4 laboratory, and inspecting and donning positive-pressure suits, entering the laboratory, moving through air pressure-resistant doors, and connecting to air-supply hoses. We will also discuss moving within and exiting the BSL-4 suit laboratories, including using the chemical shower and removing and storing positive-pressure suits.

Safety Precautions and Operating Procedures in an ABSL-4 Laboratory: 1. Biosafety Level 4 Suit Laboratory Suite Entry and Exit Procedures

Although researchers are generally knowledgeable about procedures and safety precautions required for biosafety level 1 or 2 (BSL-1/2) experiments, they may not be familiar with experimental procedures in BSL-4 suit laboratories. This article provides a detailed visual demonstration of BSL-4 suit laboratory systems check, laboratory entry, movement, and exit procedures.

אמצעי בטיחות ונהלי הפעלה בבית (א) BSL-4 מעבדה: 1. רמת בטיחות ביולוגית 4 Suit כניסה ויציאה ונהלים Suite מעבדה

Biosafety level 4 (BSL-4) suit laboratories are specifically designed to study high-consequence pathogens for which neither infection prophylaxes nor ...

Imaging Mycobacterium tuberculosis in Mice with Reporter Enzyme Fluorescence

Reporter enzyme fluorescence (REF) utilizes substrates that are specific for enzymes present in target organisms of interest for imaging or detection by fluorescence or bioluminescence. We utilize BlaC, an enzyme expressed constitutively by all M. tuberculosis strains. REF allows rapid quantification of bacteria in lungs of infected mice. The same group of mice can be imaged at many time points, greatly reducing costs, enumerating bacteria more quickly, allowing novel observations in host-pathogen interactions, and increasing statistical power, since more animals per group are readily maintained. REF is extremely sensitive due to the catalytic nature of the BlaC enzymatic reporter and specific due to the custom flourescence resonance energy transfer (FRET) or fluorogenic substrates used. REF does not require recombinant strains, ensuring normal host-pathogen interactions. We describe the imaging of M. tuberculosis infection using a FRET substrate with maximal emission at 800 nm. The wavelength of the substrate allows sensitive deep tissue imaging in mammals. We will outline aerosol infection of mice with M. tuberculosis, anesthesia of mice, administration of the REF substrate, and optical imaging. This method has been successfully applied to evaluating host-pathogen interactions and efficacy of antibiotics targeting M. tuberculosis.

Imaging Mycobacterium tuberculosis in Mice with Reporter Enzyme Fluorescence

We describe the optical imaging of mice infected with Mycobacterium tuberculosis (M. tuberculosis) using reporter enzyme fluorescence (REF). This protocol facilitates the sensitive and specific detection of M. tuberculosis in pre-clinical animal models for pathogenesis, therapeutics and vaccine research.

הדמיה שחפת Mycobacterium בעכברים עם הכתב אנזים קרינה פלואורסצנטית

Reporter enzyme fluorescence (REF) utilizes substrates that are specific for enzymes present in target organisms of interest for imaging or detection by ...

Evaluation of Zika Virus-specific T-cell Responses in Immunoprivileged Organs of Infected Ifnar1-/- Mice

The Zika virus (ZIKV) can induce inflammation in immunoprivileged organs (e.g., the brain and testis), leading to the Guillain-Barr&#233; syndrome and damaging the testes. During an infection with the ZIKV, immune cells have been shown to infiltrate into the tissues. However, the cellular mechanisms that define the protection and/or immunopathogenesis of these immune cells during a ZIKV infection are still largely unknown. Herein, we describe methods to evaluate the virus-specific T-cell functionality in these immunoprivileged organs of ZIKV-infected mice. These methods include a) a ZIKV infection and vaccine inoculation in Ifnar1-/- mice; b) histopathology, immunofluorescence, and immunohistochemistry assays to detect the virus infection and inflammation in the brain, testes, and spleen; c) the preparation of a tetramer of ZIKV-derived T-cell epitopes; d) the detection of ZIKV-specific T cells in the monocytes isolated from the brain, testes, and spleen. Using these approaches, it is possible to detect the antigen-specific T cells that have infiltrated into the immunoprivileged organs and to evaluate the functions of these T cells during the infection: potential immune protection via virus clearance and/or immunopathogenesis to exacerbate the inflammation. These findings may also help to clarify the contribution of T cells induced by the immunization against ZIKV.

Evaluation of Zika Virus-specific T-cell Responses in Immunoprivileged Organs of Infected Ifnar1-/- Mice

A protocol to evaluate antigen-specific T-cell responses in the immunoprivileged organs of the Ifnar1-/- murine model for the Zika virus (ZIKV) infection is described. This method is pivotal for investigating the cellular mechanisms of the protection and immunopathogenesis of ZIKV vaccines and is also valuable for their efficacy evaluation.

הערכה של תגובות T-cell Zika וירוס ספציפיים באיברים Immunoprivileged של עכברים נגועים Ifnar1- / -

The Zika virus (ZIKV) can induce inflammation in immunoprivileged organs (e.g., the brain and testis), leading to the Guillain-Barr&#233; syndrome and ...

Lung microRNA Profiling Across the Estrous Cycle in Ozone-exposed Mice

MicroRNA (miRNA) profiling has become of interest to researchers working in various research areas of biology and medicine. Current studies show a promising future of using miRNAs in the diagnosis and care of lung diseases. Here, we define a protocol for miRNA profiling to measure the relative abundance of a group of miRNAs predicted to regulate inflammatory genes in the lung tissue from of an ozone-induced airway inflammation mouse model. Because it has been shown that circulating sex hormone levels can affect the regulation of lung innate immunity in females, the purpose of this method is to describe an inflammatory miRNA profiling protocol in female mice, taking into consideration the estrous cycle stage of each animal at the time of ozone exposure. We also address applicable bioinformatics approaches to miRNA discovery and target identification methods using limma, an R/Bioconductor software, and functional analysis software to understand the biological context and pathways associated with differential miRNA expression.

Here we describe a method to assess lung expression of miRNAs that are predicted to regulate inflammatory genes using mice exposed to ozone or filtered air at different stages of the estrous cycle.

ריאות microRNA פרופיל לרוחב ייחום בעכברים החשופים אוזון

MicroRNA (miRNA) profiling has become of interest to researchers working in various research areas of biology and medicine. Current studies show a ...

Adoptive Immunotherapy of iNKT Cells in Glucose-6-Phosphate Isomerase (G6PI)-Induced RA Mice

Rheumatoid arthritis (RA) is a complex chronic inflammatory autoimmune disease. The pathogenesis of the disease is related to invariant natural killer T (iNKT) cells. Patients with active RA present fewer iNKT cells, defective cell function, and excessive polarization of Th1. In this study, an RA animal model was established using a mixture of hGPI325-339 and hGPI469-483 peptides. The iNKT cells were obtained by in vivo induction and in vitro purification, followed by infusion into RA mice for adoptive immunotherapy. The in vivo imaging system (IVIS) tracking revealed that iNKT cells were mainly distributed in the spleen and liver. On day 12 after cell therapy, the disease progression slowed down significantly, the clinical symptoms were alleviated, the abundance of iNKT cells in the thymus increased, the proportion of iNKT1 in the thymus decreased, and the levels of TNF-&#945;, IFN-&#947;, and IL-6 in the serum decreased. Adoptive immunotherapy of iNKT cells restored the balance of immune cells and corrected the excessive inflammation of the body.

Adoptive Immunotherapy of iNKT Cells in Glucose-6-Phosphate Isomerase G6PI-Induced RA Mice

This protocol uses G6PI mixed peptides to construct rheumatoid arthritis models that are closer to that of human rheumatoid arthritis in CD4+ T cells and cytokines. High purity invariant natural killer T cells (mainly iNKT2) with specific phenotypes and functions were obtained by in vivo induction and in vitro purification for adoptive immunotherapy.

חיסוני המאמצת של תאים iNKT ב גלוקוז-6-פוספט איזומראז (G6PI)-הנגרמת עכברים RA

Rheumatoid arthritis (RA) is a complex chronic inflammatory autoimmune disease. The pathogenesis of the disease is related to invariant natural killer T ...

Evaluation of T Follicular Helper Cells and Germinal Center Response During Influenza A Virus Infection in Mice

T Follicular Helper (Tfh) cells are an independent CD4+&#160;T cell subset specialized in providing help for germinal center (GC) development and generation of high-affinity antibodies. In influenza virus infection, robust Tfh and GC B cell responses are induced to facilitate effective virus eradication, which confers a qualified mouse model for Tfh-associated study. In this paper, we described protocols in detection of basic Tfh-associated immune response during influenza virus infection in mice. These protocols include: intranasal inoculation of influenza virus; flow cytometry staining and analysis of polyclonal and antigen-specific Tfh cells, GC B cells and plasma cells; immunofluorescence detection of GCs; enzyme-linked immunosorbent assay (ELISA) of influenza virus-specific antibody in serum. These assays basically quantify the differentiation and function of Tfh cells in influenza virus infection, thus providing help for studies in elucidating differentiation mechanism and manipulation strategy.

This paper describes protocols of evaluating Tfh and GC B response in mouse model of influenza virus infection.

הערכה של תאי עוזר T זקיקים ותגובת מרכז חיידקים במהלך שפעת זיהום וירוס בעכברים

T Follicular Helper (Tfh) cells are an independent CD4+&#160;T cell subset specialized in providing help for germinal center (GC) development and ...

Conjunctival Commensal Isolation and Identification in Mice

The ocular surface was once considered immune privileged and abiotic, but recently it appears that there is a small, but persistent commensal presence. Identification and monitoring of bacterial species at the ocular mucosa have been challenging due to their low abundance and limited availability of appropriate methodology for commensal growth and identification. There are two standard approaches: culture based or DNA sequencing methods. The first method is problematic due to the limited recoverable bacteria and the second approach identifies both live and dead bacteria leading to an aberrant representation of the ocular space. We developed a robust and sensitive method for bacterial isolation by building upon standard microbiological culturing techniques. This is a swab-based technique, utilizing an &#8220;in-lab&#8221; made thin swab that targets the lower conjunctiva, followed by an amplification step for aerobic and facultative anaerobic genera. This protocol has allowed us to isolate and identify conjunctival species such as Corynebacterium spp., Coagulase Negative Staphylococcus spp., Streptococcus spp., etc. The approach is suitable to define commensal diversity in mice under different disease conditions.

Presented here is a protocol for the isolation and amplification of aerobic and facultative anaerobic mouse conjunctival commensal bacteria using a unique eye swab and culture-based enrichment step with subsequent identification by microbiological based methods and MALDI-TOF mass spectrometry.

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The ocular surface was once considered immune privileged and abiotic, but recently it appears that there is a small, but persistent commensal presence. ...