Name: standardized preparation of single-cell suspensions from mouse lung tissue using the gentlemacs dissociator
Uploaded: 2009-07-02T00:00:00.000+00:00
Duration: 7 min 3 s
Description: Watch this Scientific Journal Video about Standardized Preparation of Single-Cell Suspensions from Mouse Lung Tissue using the gentleMACS Dissociator at JoVE.com

Miltenyi Biotec 저자 GmbH를, 독일의 직원입니다.

무균 조건 하에서 해결하려면, 그것은 층류 후드의 모든 단계를 수행하는 것이 좋습니다. 1. 자료 <ol><li> HEPES 버퍼 : 10 MM HEPES - NaOH 산도 7.4, 150 MM NaCl, 5 KCl MM, 1 MM MgCl 2, 1.8 MM CaCl 2) </li><li> Collagenase D 솔루션 : Collagenase D : 100 MG / ML (Collagenase D&gt; 0.15 U / MG) HEPES에서 버퍼 </li><li> DNase I 솔루션 : 20,000 U / ML DNase I </li><li> 산도 7.2에서 PBS 버퍼 </li><li> PEB 버퍼 : 20 부품 1 부품 BSA 주식 솔루션 Rinsing 솔루션 autoMACS </li><li> 조직 : 인접한 장기 무료 7분의 6 week 오래된 성인 여성 BALB / C 마우스에서 파생된 폐. </li><li> 옵션 : CD11c MicroBeads, 맥 MS 열, 마우스 폐 단일 셀 정지 돌기 세포의 격리 Mac 용 세퍼레이터 </li></ol> 2. 폐 조직을 Dissociating <ol><li> 적혈구를 제거하는 PBS가 포함된 배양 접시에있는 조직을 씻어. </li><li> 4.9 ML의 HEPES 버퍼를 포함하는 gentleMACS C 튜브 450 MG의 폐 조직의 최대 전송합니다. </li><li> 2 MG / ML Collagenase D.의 최종 농도 100 μL Collagenase D 솔루션 추가 </li><li> 150 MG 조직 (40 U / ML DNase I의 최종 농도) 또는 150-450 MG의 폐 조직을 20 μL DNase I (80 U / ML DNase I의 최종 농도)에 10 μL DNase I 솔루션을 추가합니다. </li><li> 단단히 C 튜브를 닫고 gentleMACS의 Dissociator에 그것을 연결합니다. 그런 다음 프로그램 &quot;m_lung_01&quot;를 실행합니다. </li><li> 37 30 분 품어 ° C 자동 또는 수동으로 회전마다 5 분 터닝과 함께. </li><li> gentleMACS의 Dissociator에 샘플을 반환하고 프로그램 &quot;m_lung_02&quot;를 실행합니다. </li></ol> 3. 여과법 <ol><li> 70 μm의 메쉬 셀 스트레이너와 뚜껑을 교체하여 필터링 세포를 수집하기 위해 50 ML 튜브를 준비합니다. </li><li> 일단 두 번째 gentleMACS 프로그램이 튜브의 샘플 자료의 배포에 따라 종료, 당신은 튜브 하단의 샘플 자료를 수집하기 위해 간단히 튜브를 원심 수 있습니다. </li><li> 적당한 1,000 μl 피펫 팁을 사용하여 C 튜브의 심장 봉인 캡을 통해 세포를 제거하고 세포 스트레이너에게 적용됩니다. </li><li> RT에서 5 ML HEPES 버퍼와 셀 스트레이너 씻으십시오. </li><li> 10 분 300xg에서 50 ML 튜브의 펠렛에 세포를 원심 분리기. </li><li> 뜨는을 기음과 PEB 버퍼의 원하는 볼륨의 세포 펠렛을 resuspend. </li></ol> 4 부 : 대표 결과 : <img src="/files/ftp_upload/1266/1266fig1.jpg" alt="그림 1" /> 그림 1. gentleMACS ™ Dissociator과 폐 분리가 92% 가능한 세포의 결과. 죽은 세포는 찬란 propidium 요오드화물의 (PI) (; 없음 PI의 얼룩 왼쪽 도트 플롯 바로 도트 플롯)와 스테인드되었습니다. <img src="/files/ftp_upload/1266/1266fig2.jpg" alt="그림 2" /> 그림 2. 가능한 leukocytes 및 내피 세포의 높은 비율의 gentleMACS의 Dissociator 결과를 사용하여 마우스 폐 조직의 해리. 파생된 단일 셀 suspensions는 CD45 - PE와 CD146 - FITC 또는 CD31 - FITC 물들일 및 유동세포계측법에 의해 분석되었다. <img src="/files/ftp_upload/1266/1266figure2c.png" alt="그림 2C" /> 그림 3. 마우스 폐 조직에서 파생된 단일 셀 정지 CD11c - FITC와 안티 mPDCA - 1 - APC 마우스 CD11c 낮은 M - PDCA - 1 + plasmacytoid의 DC가뿐만 아니라 CD11c 높은 세포를 감지하는 물들일했다. <img src="/files/ftp_upload/1266/1266fig4.jpg" alt="그림 4" /> 그림 4. CD11c MicroBeads, MiniMACS 구분 기호와 두 MS 열을 사용하여 돌기 세포의 농축.

<ol>
	<li>Swanson, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flt3-Ligand,+IL-4,+GM-CSF,+and+Adherence-Mediated+Isolation+of+Murine+Lung+Dendritic+Cells:+Assessment+of+Isolation+Technique+on+Phenotype+and+Function.">Flt3-Ligand, IL-4, GM-CSF, and Adherence-Mediated Isolation of Murine Lung Dendritic Cells: Assessment of Isolation Technique on Phenotype and Function.</a> J. Immunol. 173, 4875-4881 (2004).</li><li>Vermaelen, K., Pauwels, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accurate+and+simple+discrimination+of+mouse+pulmonary+dendritic+cell+and+macrophage+populations+by+flow+cytometry:+Methodology+and+new+insights.">Accurate and simple discrimination of mouse pulmonary dendritic cell and macrophage populations by flow cytometry: Methodology and new insights.</a> Cytometry Part A. 61A, 170-177 (2004).</li></ol>

The authors are employees of Miltenyi Biotec GmbH who make the instrument used in this article.

이 비디오에서는, 우리는 마우스 폐 조직의 분리를위한 새로운 방법을 소개합니다. 저희는 폐 조직의 기계적 효소 치료의 조합이 가능한 leukocytes 및 내피 세포의 높은 비율을 알아낼 것을 보여줍니다. 특히, 조직의 기계적 붕괴는 gentleMACS의 Dissociator에 의해 달성되었다. 고정자 시스템, 부드러운 방식으로 dissociates 조직 - gentleMACS C 튜브는 로터가 포함되어 있습니다. 절차는 악기의 프로그램 설정에 의해 제어됩니다. 설정은 세포의 높은 수율과 생존을 달성하기 위해 최적화되었다. 파생된 단일 셀 suspensions는 CD45 - PE와 CD146 - FITC 또는 CD31 - FITC 물들일 및 leukocytes과 내피 세포를 감지하는 유동세포계측법에 의해 분석되었다. 단일 세포 현탁액에서 돌기 세포는 CD11c - FITC와 안티 mPDCA - 1 - APC 물들일되었습니다. 또한, 마우스 폐 단일 셀 정지 돌기 세포는 맥 기술을 사용하여 풍부하게되었습니다. 돌기 세포는 자기 (磁气) CD11c MicroBeads를 사용하여 분류하고 MiniMACS 구분 기호와 MS 열을 사용하여 분리되었다 일반적으로 1,2와 새로운 분리 프로토콜의 결합 자기 셀 정렬은 폐 조직의 특정 세포 유형을 얻을 수있는 표준 방법을 나타냅니다.

<table><tbody><tr><td>C Tubes</td><td>Miltenyi Biotec</td><td>Order no. 130-093-237</td><td>&lt;a href=&quot;http://www.miltenyibiotec.com/en/PG_709_765_gentleMACS_C_Tubes.aspx&quot;&gt;http://www.miltenyibiotec.com/en/PG_709_765_gentleMACS_C_Tubes.aspx&lt;/a&gt;</td></tr><tr><td>gentleMACS&amp;#8482; Starting Kit</td><td>Miltenyi Biotec</td><td>Order no. 130-093-235</td><td>&lt;a href=&quot;http://www.miltenyibiotec.com/en/PG_709_763_gentleMACS_Dissociator.aspx&quot;&gt;http://www.miltenyibiotec.com/en/PG_709_763_gentleMACS_Dissociator.aspx&lt;/a&gt;</td></tr><tr><td>MACSmix&amp;#8482; Tube Rotator</td><td>Miltenyi Biotec</td><td>Order no. 130-090-753</td><td>&lt;a href=&quot;http://www.miltenyibiotec.com/en/PG_34_251_MACSmix_Tube_Rotator.aspx&quot;&gt;http://www.miltenyibiotec.com/en/PG_34_251_MACSmix_Tube_Rotator.aspx&lt;/a&gt;</td></tr><tr><td>autoMACS&amp;#8482; Rinsing Solution</td><td>Miltenyi Biotec</td><td>Order no. 130-091-222</td><td>&lt;a href=&quot;http://www.miltenyibiotec.com/en/PG_134_155_autoMACS_Rinsing_Solution.aspx&quot;&gt;http://www.miltenyibiotec.com/en/PG_134_155_autoMACS_Rinsing_Solution.aspx&lt;/a&gt;</td></tr><tr><td>MACS&amp;#174; BSA Stock Solution</td><td>Miltenyi Biotec</td><td>Order no. 130-091-376</td><td>&lt;a href=&quot;http://www.miltenyibiotec.com/en/PG_33_295_MACS_BSA_Stock_Solution.aspx&quot;&gt;http://www.miltenyibiotec.com/en/PG_33_295_MACS_BSA_Stock_Solution.aspx&lt;/a&gt;</td></tr><tr><td>CD11c MicroBeads</td><td>Miltenyi Biotec</td><td>Order No. 130-052-001</td><td>&lt;a href=&quot;http://www.miltenyibiotec.com/en/PG_93_132_CD11c_MicroBeads.aspx&quot;&gt;http://www.miltenyibiotec.com/en/PG_93_132_CD11c_MicroBeads.aspx&lt;/a&gt;</td></tr><tr><td>CD11c-FITC, CD11c-PE</td><td>Miltenyi Biotec</td><td>Order no. 130-092-026130-091-830</td><td>&lt;a href=&quot;http://www.miltenyibiotec.com/en/PG_599_329_CD11c.aspx&quot;&gt;http://www.miltenyibiotec.com/en/PG_599_329_CD11c.aspx&lt;/a&gt;</td></tr><tr><td>Anti-mPDCA-1-APC</td><td>Miltenyi Biotec</td><td>Order no. 130-091-963</td><td>&lt;a href=&quot;http://www.miltenyibiotec.com/en/PG_599_325_Anti_mPDCA_1.aspx&quot;&gt;http://www.miltenyibiotec.com/en/PG_599_325_Anti_mPDCA_1.aspx&lt;/a&gt;</td></tr><tr><td>CD146-FITC</td><td>Miltenyi Biotec</td><td>Order no. 130-092-026</td><td>&lt;a href=&quot;http://www.miltenyibiotec.com/en/PG_599_417_CD146_LSEC_.aspx&quot;&gt;http://www.miltenyibiotec.com/en/PG_599_417_CD146_LSEC_.aspx&lt;/a&gt;</td></tr><tr><td>CD45-PE</td><td>Miltenyi Biotec</td><td>Order no. 130-091-610</td><td>&lt;a href=&quot;http://www.miltenyibiotec.com/en/PG_599_331_CD45.aspx&quot;&gt;http://www.miltenyibiotec.com/en/PG_599_331_CD45.aspx&lt;/a&gt;</td></tr><tr><td>Propidium Iodide Solution</td><td>Miltenyi Biotec</td><td>Order no. 130-093-233</td><td>&lt;a href=&quot;http://www.miltenyibiotec.com/en/PG_337_744_Propidium_Iodide_Solution.aspx&quot;&gt;http://www.miltenyibiotec.com/en/PG_337_744_Propidium_Iodide_Solution.aspx&lt;/a&gt;</td></tr><tr><td>Art 1000 Reach Pipet Tips</td><td>Molecular BioProducts</td><td>Order no. 2079</td><td>&lt;a href=&quot;http://www.mbpinc.com/ASP/products.aspx?query_TipID=61&quot;&gt;http://www.mbpinc.com/ASP/products.aspx?query_TipID=61&lt;/a&gt;&lt;br/&gt;&lt;a href=&quot;http://www.mbpinc.com/html/products/art/display.html&quot;&gt;http://www.mbpinc.com/html/products/art/display.html&lt;/a&gt;</td></tr></tbody></table>

standardized preparation of single-cell suspensions from mouse lung tissue using the gentlemacs dissociator

조직에서 단일 셀 suspensions의 준비 세포 연구에 많은 실험을위한 중요한 사전 요구 사항에 해당됩니다. 전체 장기 dissociating의 과정은 재현성 방식으로 가능한 세포의 높은 숫자를 얻기 위해서는 특정 매개 변수를 필요로합니다. gentleMACS Dissociator은 간단하고 실용적인 프로토콜이 작업을 최적화합니다. 악기는 마우스를 포함하여 폐 조직 유형의 다양한 효율적이지만 부드러운 분리에 최적화된 미리 프로그램된 설정이 포함되어 있습니다. 이 간행물의 생쥐에서 파생된 폐 조직에 gentleMACS의 Dissociator의 사용을 시연합니다.

Watch this Scientific Journal Video about Standardized Preparation of Single-Cell Suspensions from Mouse Lung Tissue using the gentleMACS Dissociator at JoVE.com

Standardized Preparation of Single-Cell Suspensions from Mouse Lung Tissue using the gentleMACS Dissociator

특정 조직 유형의 세포를 Dissociating하면 조직의 동요에 대한 특정 매개 변수가 가능한, culturable 세포의 높은 볼륨을 얻기 위해 필요합니다. Miltenyi gentleMACS dissociator은 간단하고 실용적인 프로토콜이 작업을 최적화합니다. 이 발행물에서 폐 조직에서이 장치의 사용이 설명되어 있습니다.

gentleMACS Dissociator를 사용하여 마우스 폐 조직에서 단일 셀 Suspensions의 표준화 준비

The preparation of single-cell suspensions from tissues is an important prerequisite for many experiments in cellular research. The process of ...

standardized-preparation-single-cell-suspensions-from-mouse-lung

Universidad San Sebastian

Research

JoVE Journal

Biology

34.5K 조회수.  Miltenyi Biotec,GmbH.  中文 日本語 

비디오: Standardized Preparation of Single-Cell Suspensions from Mouse Lung Tissue using the gentleMACS Dissociator

Isolation of Mouse Lung Dendritic Cells

Lung dendritic cells (DC) play a fundamental role in sensing invading pathogens 1,2 as well as in the control of tolerogenic responses 3 in the respiratory tract. At least three main subsets of lung dendritic cells have been described in mice: conventional DC (cDC) 4, plasmacytoid DC (pDC) 5 and the IFN-producing killer DC (IKDC) 6,7. The cDC subset is the most prominent DC subset in the lung 8. 
The common marker known to identify DC subsets is CD11c, a type I transmembrane integrin (&beta;2) that is also expressed on monocytes, macrophages, neutrophils and some B cells 9. In some tissues, using CD11c as a marker to identify mouse DC is valid, as in spleen, where most CD11c+ cells represent the cDC subset which expresses high levels of the major histocompatibility complex class II (MHC-II). However, the lung is a more heterogeneous tissue where beside DC subsets, there is a high percentage of a distinct cell population that expresses high levels of CD11c bout low levels of MHC-II. Based on its characterization and mostly on its expression of F4&#47;80, an splenic macrophage marker, the CD11chiMHC-IIlo lung cell population has been identified as pulmonary macrophages 10 and more recently, as a potential DC precursor 11. 
In contrast to mouse pDC, the study of the specific role of cDC in the pulmonary immune response has been limited due to the lack of a specific marker that could help in the isolation of these cells. Therefore, in this work, we describe a procedure to isolate highly purified mouse lung cDC. The isolation of pulmonary DC subsets represents a very useful tool to gain insights into the function of these cells in response to respiratory pathogens as well as environmental factors that can trigger the host immune response in the lung.

A highly purified preparation of mouse lung dendritic cells is described. Specific emphasis is given to the isolation of conventional dendritic cell subset.

마우스 폐에 돌기 세포의 분리

Lung dendritic cells (DC) play a fundamental role in sensing invading pathogens 1,2 as well as in the control of tolerogenic responses 3 in the ...

연구

면역학 및 감염병학

Preparation of Single-Cell Suspensions from Mouse Spleen with the gentleMACS Dissociator

Single-cell suspensions are a prerequisite for experiments in cell separation, cell analysis and cell culture.  To avoid tedious and often painful manual dissociations the gentleMACS Dissociator allows one to dissociate tissue very efficiently under controlled and reproducible conditions.  The gentleMACS Dissociator can optimally dissociate mouse spleen, combining timesaving and standardization with user-safety.

This video describes how to dissociate mouse spleens using the gentleMACS Dissociator, an automated bench-top device that can mechanically disrupt tissues using special tubes to produce viable cell suspensions. Following dissociation, spleens are filtered, centrifuged, and resuspended for further applications.

This video describes a simple, time-saving technique for automated tissue dissociation using the gentleMACS Dissociator to prepare single-cell suspensions of mouse splenocytes.

gentleMACS Dissociator과 마우스 비장에서 단일 셀 Suspensions의 준비

Single-cell suspensions are a prerequisite for experiments in cell separation, cell analysis and cell culture.  To avoid tedious and often painful manual ...

생물학

Generation of Single-Cell Suspensions from Mouse Neural Tissue

Within the nervous system, hundreds of neuronal and glial cell types have been described. Each specific cell type in the brain or spinal cord has a repertoire of cell surface molecules, or molecular determinants, through which it can be identified and characterized. Currently, robust cell identification and separation technologies require single-cell preparations to be generated while simultaneously limiting cell death and destruction of characteristic surface protein. The gentleMACS Dissociator, when used in combination with trypsin or papain-based dissociation kits, can effectively and gently dissociate brain tissue while preserving antigen epitopes and limiting cell loss. Standardized preparation of single-cell suspensions is achieved using C Tubes and optimized, preset gentleMACS Programs. Once generated, single-cell suspensions can be treated with monoclonal conjugates like Anti-Prominin-1 MicroBeads, which identify neural progenitors, or purified further using Myelin Removal Beads.

Dissociating cells from specific tissue types requires specific parameters for tissue aggitation to obtain a high volume of viable, culturable cells. The Miltenyi gentleMACS Dissociator optimizes this task with a simple, practical protocol. In this publication the use of this apparatus on nerual tissue is explained.

마우스 신경 조직에서 단일 셀 Suspensions의 생성

Within the nervous system, hundreds of neuronal and glial cell types have been described. Each specific cell type in the brain or spinal cord has a ...

Isolation and In Vitro Culture of Murine and Human Alveolar Macrophages

Alveolar macrophages are terminally differentiated, lung-resident macrophages of prenatal origin. Alveolar macrophages are unique in their long life and their important role in lung development and function, as well as their lung-localized responses to infection and inflammation. To date, no unified method for identification, isolation, and handling of alveolar macrophages from humans and mice exists. Such a method is needed for studies on these important innate immune cells in various experimental settings. The method described here, which can be easily adopted by any laboratory, is a simplified approach to harvesting alveolar macrophages from bronchoalveolar lavage fluid or from lung tissue and maintaining them in vitro. Because alveolar macrophages primarily occur as adherent cells in the alveoli, the focus of this method is on dislodging them prior to harvest and identification. The lung is a highly vascularized organ, and various cell types of myeloid and lymphoid origin inhabit, interact, and are influenced by the lung microenvironment. By using the set of surface markers described here, researchers can easily and unambiguously distinguish alveolar macrophages from other leukocytes, and purify them for downstream applications. The culture method developed herein supports both human and mouse alveolar macrophages for in vitro growth, and is compatible with cellular and molecular studies.

Isolation and In Vitro Culture of Murine and Human Alveolar Macrophages

This communication describes methodologies for isolation and culture of alveolar macrophages from humans and murine models for experimental purposes.

고립과 Murine 인간과 치경 대 식 세포의 생체 외에서 문화

Alveolar macrophages are terminally differentiated, lung-resident macrophages of prenatal origin. Alveolar macrophages are unique in their long life and ...

Isolation &#38; Characterization of Hoechstlow CD45negative Mouse Lung Mesenchymal Stem Cells

Tissue resident mesenchymal stem cells (MSC) are important regulators of tissue repair or regeneration, fibrosis, inflammation, angiogenesis and tumor formation. Taken together these studies suggest that resident lung MSC play a role during pulmonary tissue homeostasis, injury and repair during diseases such as pulmonary fibrosis (PF) and arterial hypertension (PAH). Here we describe a technology to define a population of resident lung MSC. The definition of this population in vivo pulmonary tissue using a define set of markers facilitates the repeated isolation of a well-characterized stem cell population by flow cytometry and the study of a specific cell type and function.

Isolation & Characterization of Hoechstlow CD45negative Mouse Lung Mesenchymal Stem Cells

In this article we demonstrate the isolation of murine resident lung mesenchymal stem cells (lung MSC), their expansion, characterization and analysis of immunomodulatory properties.

Hoechst의 분리 및 특성 낮은 CD45 부정적인 마우스 폐 Mesenchymal 줄기 세포

Tissue resident mesenchymal stem cells (MSC) are important regulators of tissue repair or regeneration, fibrosis, inflammation, angiogenesis and tumor ...

단일 세포 분석을 위한 반자동 기계-효소 조직 분리

Mechanical Dissociation of Tissues for Single Cell Analysis Using a Motorized Device

Being able to isolate and prepare single cells for the analysis of tissue samples has rapidly become crucial for new biomedical discoveries and research. Manual protocols for single-cell isolations are highly time-consuming and prone to user variability. Automated mechanical protocols are able to reduce processing time and sample variability but aren't easily accessible or cost-effective in lower-resourced research settings. The device described here was designed for semi-automated tissue dissociation using commercially available materials as a low-cost alternative for academic laboratories. Instructions to fabricate, assemble, and operate the device design have been provided. The dissociation protocol reliably produces single-cell suspensions with comparable cell yields and sample viability to manual preparations across multiple mouse tissues. The protocol provides the ability to process up to 12 tissue samples simultaneously per device, making studies requiring large sample sizes more manageable. The accompanying software also allows for customization of the device protocol to accommodate varying tissues and experimental constraints.

저자 스포트라이트: 약물 전달 및 면역 요법을 위한 나노 입자 기술의 발전

A general protocol for the combined enzymatic and semi-automated mechanical dissociation of tissues to generate single-cell suspensions for downstream analyses, such as flow cytometry, is provided. Instructions for the fabrication, assembly, and operation of the low-cost mechanical device developed for this protocol are included.

전동 장치를 사용한 단일 세포 분석을 위한 조직의 기계적 해리

Being able to isolate and prepare single cells for the analysis of tissue samples has rapidly become crucial for new biomedical discoveries and research. ...

생체공학

Isolation and Characterization of Antigen-Specific T cells in Mouse Lungs(마우스 폐에서 항원 특이적 T 세포의 분리 및 특성 분석)

Identification of Rare Antigen-Specific T Cells from Mouse Lungs with Peptide:Major Histocompatibility Complex Tetramers

The identification and characterization of antigen-specific T cells during health and disease remains a key to improving our understanding of immune pathophysiology. The technical challenges of tracking antigen-specific T cell populations within the endogenous T cell repertoire have been greatly advanced by the development of peptide:MHC tetramer reagents. These fluorescently labeled soluble multimers of MHC class I or class II molecules complexed to antigenic peptide epitopes bind directly to T cells with corresponding T cell receptor (TCR) specificity and can, therefore, identify antigen-specific T cell populations in their native state without a requirement for a functional response induced by ex vivo stimulation. For exceedingly rare populations, tetramer-bound T cells can be magnetically enriched to increase the sensitivity and reliability of detection.
As the investigation of tissue-resident T cell immunity deepens, there is a pressing need to identify antigen-specific T cells that traffic to and reside in nonlymphoid tissues. In this protocol, we present a detailed set of instructions for the isolation and characterization of antigen-specific T cells present within mouse lungs. This involves the isolation of T cells from digested lung tissue followed by a general T cell magnetic enrichment step and tetramer staining for flow cytometry analysis and sorting. The steps highlighted in this protocol utilize common techniques and readily available reagents, making it accessible for nearly any researcher engaged in mouse T cell immunology, and are highly adaptable for a variety of downstream analyses of any low frequency antigen-specific T cell population residing within the lungs.

저자 스포트라이트: Tetramers를 사용하여 마우스 폐에서 항원 특이적 T 세포를 검출하기 위한 최적화된 프로토콜

We provide a detailed protocol for isolating and identifying rare antigen-specific T cell populations in mouse lungs through magnetic bead-based T cell enrichment and peptide:major histocompatibility complex (MHC) tetramers.

펩타이드를 이용한 생쥐 폐의 희귀 항원 특이적 T 세포 식별: 주요 조직적합성 복합체 테트라머

The identification and characterization of antigen-specific T cells during health and disease remains a key to improving our understanding of immune ...

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펩타이드를 이용한 생쥐 폐의 희귀 항원 특이적 T 세포 식별: 주요 조직적합성 복합체 테트라머

펩타이드를 이용한 생쥐 폐의 희귀 항원 특이적 T 세포 식별: 주요 조직적합성 복합체 테트라머