EB, KMB, 그리고 HX 동일하게 기여하고 있습니다. 이 연구는 위장 암 P50 CA062924, NIH K23 CA148964, Lustgarten 재단, Viragh 재단, ASCO 젊은 연구자 상, NIH 5T32 CA0090701-28 교육 교부금, 솔 골드만 삭스의 췌장암 센터, 국립 췌장 재단에서 NCI의 포자에 의해 지원되었다.

B7-H1/PD-L1, 면역 규제 세포 표면 단백질의 B7 가족의 일원은 (PD 프로그래밍 사망-1, 그 수용체와의 상호 작용을 통해 세포 매개 면역 반응의 부정적인 규제에 중요한 역할을 -1) 1,2. 종양 세포의 B7-H1의 Overexpression는 흑색 증, glioblastoma, 그리고 폐, 유방, 대장, 난소 및 신장 세포의 carcinomas을 포함 인간 암의 숫자에 기록되었으며, 항 종양 저해하는 표시되었습니다 T를 세포 면역 3-8. 최근 췌장 선암의 조직에 의해 B7-H1 표현은 잠재적 인 전조 마커 9,10로 확인되었습니다. 또한 췌장암의 마우스 모델에서 B7-H1의 봉쇄는 안티 종양 응답 11 생산 보여왔다. 이러한 데이터는 잠재적 인 치료 목표로 B7-H1의 중요성을 권장합니다. 안티 - B7-H1의 봉쇄 항체 따라서 여러 인간에 대한 임상 실험에서 테스트되고 있습니다흑색 증과 폐, 대장, 신장, 위장, 췌장 (12)의 암 등의 고체 종양. 결국 치료를 타겟으로 B7-H1의 혜택을받을 환자를 식별 할 수 있도록 위해서는 표현과 B7-H1과 B7-H1의 봉쇄 항체 치료에 대한 환자 반응의 국산화 사이의 상관 관계를 조사하는 것이 중요합니다. immunohistochemistry를 통해 인간의 췌장 선암의 조직에서 B7-H1의 표현을 조사하면이 공동 억제 신호 분자가 종양의 microenvironment에 antitumor 면역의 억제에 기여하는 방법에 대한 이해를 제공 할 것입니다. 첸과 동료에 의해 ​​개발 된 안티 - B7-H1 단클론 항체 (클론 5H1)가 인간의 neoplastic 조직 4,8의 여러 유형의 cryosections에서 신뢰할 수있는 염색 결과를 생산하기 위해 표시하지만, 파라핀 - 임베디드 슬라이드에 얼룩되었습니다 것은 때까지 도전이었다 최근 13-18. 우리는 B7-H1 staini을 개발했습니다췌장 선암의 조직의 파라핀 - 임베디드 슬라이드에 대한 것이다 프로토콜. 여기에 설명 된 B7-H1의 착색 프로토콜은 약간의 배경 B7-H1의 일관성 막과 세포질 염색을 생산하고 있습니다. 1. 드 - parafinization <ol><li> 구워 20 분에 55 ° C에서 슬라이드. </li><li> 10 분을위한 크실렌의 화학 후드에서 잠그다는 슬라이드. </li><li> 10 분에 대한 새로운 크실렌에 젖어 슬라이드. </li><li> 젖어은 5 분 동안 100 %의 에탄올에 슬라이드. </li><li> 빠져 5 분을위한 신선한 100 % 에탄올에 슬라이드. </li><li> 젖어은 5 분 동안 95 %의 에탄올에 슬라이드. </li><li> 젖어은 5 분 동안 80 % 에탄올에 슬라이드. </li><li> 젖어은 5 분 동안 탈 이온수에 슬라이드. </li></ol> 2. 항원 불러 오기 <ol><li> 그리고 125 ° C 30 초를위한 트리스 - EDTA (에틸렌 다이아 민 테트라 초산) 버퍼 (산도 9.0)과 압력 밥솥의 열, 90 ° 10 초에 C에 젖어 슬라이드. </li><li> 60 분에 슬라이드가 멋진 보자. </li><li> 5 분에 TBST에 빠져 슬라이드, repe두 번에. </li></ol> 3. 더럽히는 것 <ol><li> 슬라이드에 조직 주위에 솔루션을 닦아. 소수성 펜으로 조직 주위에 원을 표시합니다. </li><li> 습도 챔버에 슬라이드를 삽입하고 전체 염색 과정에서 슬라이드의 조직의 건조를 방지. 5 분에 퍼 옥시 데이즈 블록에서 슬라이드 및 품다 슬라이드에 소수성 원에서 퍼 옥시 데이즈 블록의 장소 한 방울 (완전히 조직 영역을 커버하기 위해 약 200 μl 이상). </li><li> 5 분에 TBST에서 발생 후, TBST로 슬라이드를 씻어. </li><li> BLOCK ACE 장소 한 방울로 15 분 각 슬라이드 및 배양에 버퍼를 차단. </li><li> 5 분에 TBST에서 발생 후, TBST로 슬라이드를 씻어. </li><li> 15 분 각 슬라이드 및 배양에 아비딘 솔루션의 장소 한 방울. </li><li> 5 분에 TBST에서 발생 후, TBST로 슬라이드를 씻어. </li><li> 15 분 각 슬라이드 및 배양에 비오틴 솔루션의 장소 한 방울. </li><li> SLI를 씻어TBST로 DES, 5 분에 TBST에서 다음 장소. </li><li> 마우스 방지 B7-H1 단클론 항체의 1:1,000 희석 (복제 5H1) 또는 마우스 IgG1과 함께 슬라이드를 배양 4에 200 μl TBST에서 κ isotype 제어 ° C 22 시간 + / 밤새 - 1 시간. TBST의 최종 항체 농도는 2.57 μg / ML입니다. </li><li> 5 분에 TBST에서 발생 후, TBST로 슬라이드를 씻어. 두 번 반복합니다. </li><li> 30 분의 1:500 농도에서 비오틴 복합 쥐 안티 - 마우스 IgG1 항체 보조 (200 μl 2 % 염소 혈청 솔루션)에 슬라이드를 품다. </li><li> 5 분에 TBST에서 발생 후, TBST로 슬라이드를 씻어. 두 번 반복합니다. </li><li> Streptavidin-비오틴 복합체 대신 한 방울은 15 분 각 슬라이드 및 배양에 (사용 전에 적어도 30 분을 준비해야합니다.) </li><li> 5 분에 TBST에서 발생 후, TBST로 슬라이드를 씻어. 두 번 반복합니다. </li><li> 4 분 각 슬라이드 및 배양에 증폭 시약 대신 한 방울 (TBST로 2시 1분을 희석). </li><li> T와 슬라이드를 씻어영국 서머 타임, 5 분에 TBST에서 다음 장소. 두 번 반복합니다. </li><li> 15 분 각 슬라이드 및 배양에 Streptavidin-HRP 대신 한 방울. </li><li> 5 분에 TBST에서 발생 후, TBST로 슬라이드를 씻어. 두 번 반복합니다. </li><li> 각 슬라이드에 하나 3,3-Diaminobenzidine의 드롭 (DAB) 기판과 chromogen을 놓고 2 분의 색상을 개발합니다. </li><li> 즉시 색상 개발 한 후, 1-2 분 동안 ddH 2 O로 슬라이드를 씻으십시오. </li><li> 90 초 동안 hematoxylin 한 방울과 슬라이드를 품다. </li><li> 1-2 분 동안 ddH 2 O로 씻으십시오. </li><li> 1-2 분 동안 비누의 수돗물로 씻으십시오. </li><li> 1-2 분 동안 ddH 2 O로 씻으십시오. </li></ol> 4. 탈수 및 장착 <ol><li> 3 분에 80 % 에탄올에 슬라이드를 빠져. </li><li> 3 분에 95 %의 에탄올에 슬라이드를 빠져. </li><li> 5 분을위한 100 % 에탄올에 슬라이드를 빠져. 반복합니다. </li><li> 5 분에 크실렌에서 슬라이드를 빠져. 반복합니다. </li><li> 한 방울을 추가슬라이드과 장소 coverslip에 Cytoseal 60. </li></ol> 표 1은 위에서 설명한 절차에 사용되는 시약을 나열합니다.

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L., Chen, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Colocalization+of+inflammatory+response+with+B7-h1+expression+in+human+melanocytic+lesions+supports+an+adaptive+resistance+mechanism+of+immune+escape.">Colocalization of inflammatory response with B7-h1 expression in human melanocytic lesions supports an adaptive resistance mechanism of immune escape.</a> Sci. Transl. Med. 4, 127ra37 (2012).</li></ol>

관심 없음 충돌이 선언 없습니다.

cryosections에서 B7-H1의 Immunohistochemical 착색은 암 3,4의 다양한 사전 연구에서보고되었습니다. 그러나, 임상 종양 샘플을 일반적으로 파라핀 - 임베디드 블록의 형태로 사용할 수 있습니다. 5H1 단클론 항체는 최근 성공적으로 신장 세포 암종, 자궁 경부 암, 방광 암, 흑색 증 13-18 등의 파라핀 포함 된 종양 조직의 착색에 적용되었습니다. 파라핀 - 임베디드 9,19 또는 그라 이오 ?...

<table border="1"><tbody><tr><td> 시약의 이름 </td><td> 회사 </td><td> 카탈로그 번호 </td><td> 코멘트 </td></tr><tr><td> CSA 시스템 </td><td> Dako </td><td> K1500 </td><td> 퍼 옥시 데이즈 블록, streptavidin-비오틴 단지, 증폭 시약, streptavidin-HRP 및 DAB 기판과 chromogen 다음 프로토콜에 언급 된 시약을 포함 </td></tr><tr><td> 아비딘 / 비오틴 차단 키트 </td><td> 벡터 </td><td> SP-2001 </td><td></td></tr><tr><td> 차단 ACE </td><td> Serotec </td><td> BUF029 </td><td></td></tr><tr><td> 비오틴 안티 - 마우스 IgG1 </td><td> BD </td><td> 553441 </td><td></td></tr><tr><td> 마우스 IgG1 K Isotype 제어 </td><td> eBioscience </td><td> 14-4714-85 </td><td></td></tr><tr><td> 10X TBS (생리 트리스 버퍼) </td><td> Cellgro </td><td> 46-012-CM </td><td> 10xTBS 80 G / L NaCl 및 24.2 g / L Tirs을 포함, 세차를위한 ddH2O와 1X로 희석; </td></tr><tr><td> 십대 초반 20 </td><td> 시그마 - 알드리치 </td><td> P1379 </td><td> 한 ML은 TBST를 만들기 위해 1X TBS의 1 L에 추가 </td></tr><tr><td> 대상 검색, 20x </td><td> Celerus 웨이브 </td><td> 014-3000-000 </td><td> ddH 2 O로 1 회에 희석 </td></tr><tr><td> 190 증명 에틸 알코올 </td><td> Pharmco - Aaper </td><td> 111000190 </td><td></td></tr><tr><td> 200 증명 에틸 알코올 </td><td> Pharmo - Aaper </td><td> 111000200 </td><td></td></tr><tr><td> 크실렌 </td><td> VWR </td><td> 95057-827 </td><td></td></tr><tr><td> Hematoxylin </td><td> 리차드 - 앨런 과학 </td><td> 72,804 </td><td></td></tr><tr><td> Cytoseal 60 </td><td> 리차드 - 앨런 과학 </td><td> 8310-4 </td><td></td></tr><tr><td> Coverslips </td><td> 코닝 </td><td> 2940-245 </td><td></td></tr><tr><td> 습도 챔버 </td><td> 시그마 </td><td> H6644 </td><td></td></tr><tr><td></td><td></td><td></td><td> 표 1. 시약 및 장비. </td></tr></tbody></table>

immunohistochemical staining of b7-h1 (pd-l1) on paraffin-embedded slides of pancreatic adenocarcinoma tissue

B7-H1/PD-L1, 면역 규제 세포 표면 단백질의 B7 가족의 일원은 (PD 프로그래밍 사망-1, 그 수용체와의 상호 작용을 통해 세포 매개 면역 반응의 부정적인 규제에 중요한 역할을 -1) 1,2. 종양 세포의 B7-H1의 Overexpression는 흑색 증, glioblastoma, 그리고 폐, 유방, 대장, 난소 및 신장 세포의 carcinomas을 포함 인간 암의 숫자에 기록되었으며, 항 종양 저해하는 표시되었습니다 T를 세포 면역 3-8. 최근 췌장 선암의 조직에 의해 B7-H1 표현은 잠재적 인 전조 마커 9,10로 확인되었습니다. 또한 췌장암의 마우스 모델에서 B7-H1의 봉쇄는 안티 종양 응답 11 생산 보여왔다. 이러한 데이터는 잠재적 인 치료 목표로 B7-H1의 중요성을 권장합니다. 안티 - B7-H1의 봉쇄 항체 따라서 mul에 대한 임상 실험에서 테스트되고 있습니다흑색 증과 폐, 대장, 신장, 위장, 췌장 (12)의 암을 포함 tiple 인간의 고체 종양. 결국 치료를 타겟으로 B7-H1의 혜택을받을 환자를 식별 할 수 있도록 위해서는 표현과 B7-H1과 B7-H1의 봉쇄 항체 치료에 대한 환자 반응의 국산화 사이의 상관 관계를 조사하는 것이 중요합니다. immunohistochemistry를 통해 인간의 췌장 선암의 조직에서 B7-H1의 표현을 조사하면이 공동 억제 신호 분자가 종양의 microenvironment에 antitumor 면역의 억제에 기여하는 방법에 대한 이해를 제공 할 것입니다. 첸과 동료에 의해 ​​개발 된 안티 - B7-H1 단클론 항체 (클론 5H1)가 인간의 neoplastic 조직 4,8의 여러 유형의 cryosections에서 신뢰할 수있는 염색 결과를 생산하기 위해 표시하지만, 파라핀 - 임베디드 슬라이드에 얼룩되었습니다 것은 때까지 도전이었다 최근 13-18. 우리는 개발자가췌장 선암의 조직의 파라핀 - 임베디드 슬라이드에 대한 사랑의 도피를 떠났을 B7-H1의 착색 프로토콜. 여기에 설명 된 B7-H1의 착색 프로토콜은 약간의 배경 B7-H1의 일관성 막과 세포질 염색을 생산하고 있습니다.

B7-H1의 성공적인 immunohistochemical 얼룩은 췌장 선암의 B7-H1 (갈색 신호 DAB 색상 개발)의 이기종 표현을 보여줍니다. 다른 B7-H1 또는 B7-H1 (도 2 및도 3)의 작은 표현 중 하나 주로 세포질 표현을 할 수 있지만 일부 췌장 선암 세포는 B7-H1의 주로 막 표현 (그림 1A)를 갖추고 있습니다. 일반 췌장 덕트 상피는 (그림 4) B7-H1 약간을 표현한다. 마우스 IgG1 κ isotype 제어 항체?...

Watch this Scientific Journal Video about Immunohistochemical Staining of B7-H1 PD-L1 on Paraffin-embedded Slides of Pancreatic Adenocarcinoma Tissue at JoVE.com

Immunohistochemical Staining of B7-H1 PD-L1 on Paraffin-embedded Slides of Pancreatic Adenocarcinoma Tissue

B7-H1 (PD-L1) 및 PD-1의 바인딩은 종양의 microenvironment의 주요 종양 유발 면역 신호를 제공합니다. 표현과 췌장 선암의 B7-H1의 국산화를 특성화 할 수 immunohistochemical 얼룩 기법이 여기에 설명되어 있습니다.

췌장 선암 조직의에서 B7-H1의 Immunohistochemical 착색 (PD-L1) 파라핀은 - 임베디드 슬라이드

B7-H1/PD-L1, a member  of the B7 family of immune-regulatory cell-surface proteins, plays an important  role in the negative regulation of cell-mediated ...

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Immunohistochemical Staining of B7-H1 (PD-L1) on Paraffin-embedded Slides of Pancreatic Adenocarcinoma Tissue

23.0K 조회수.  The Johns Hopkins University School of Medicine. B7-H1 (PD-L1) 및 PD-1의 바인딩은 종양의 microenvironment의 주요 종양 유발 면역 신호를 제공합니다. 표현과 췌장 선암의 B7-H1의 국산화를 특성화 할 수 immunohistochemical 얼룩 기법이 여기에 설명되어 있습니다.

비디오: Immunohistochemical Staining of B7-H1 PD-L1 on Paraffin-embedded Slides of Pancreatic Adenocarcinoma Tissue

Staining Protocols for Human Pancreatic Islets

Estimates of islet area and numbers and endocrine cell composition in the adult human pancreas vary from several hundred thousand to several million and beta mass ranges from 500 to 1500 mg 1-3. With this known heterogeneity, a standard processing and staining procedure was developed so that pancreatic regions were clearly defined and islets characterized using rigorous histopathology and immunolocalization examinations. 
Standardized procedures for processing human pancreas recovered from organ donors are described in part 1 of this series. The pancreas is processed into 3 main regions (head, body, tail) followed by transverse sections. Transverse sections from the pancreas head are further divided, as indicated based on size, and numbered alphabetically to denote subsections. This standardization allows for a complete cross sectional analysis of the head region including the uncinate region which contains islets composed primarily of pancreatic polypeptide cells to the tail region.
The current report comprises part 2 of this series and describes the procedures used for serial sectioning and histopathological characterization of the pancreatic paraffin sections with an emphasis on islet endocrine cells, replication, and T-cell infiltrates. Pathology of pancreatic sections is intended to characterize both exocrine, ductular, and endocrine components. The exocrine compartment is evaluated for the presence of pancreatitis (active or chronic), atrophy, fibrosis, and fat, as well as the duct system, particularly in relationship to the presence of pancreatic intraductal neoplasia4. Islets are evaluated for morphology, size, and density, endocrine cells, inflammation, fibrosis, amyloid, and the presence of replicating or apoptotic cells using H&amp;E and IHC stains. 
The final component described in part 2 is the provision of the stained slides as digitized whole slide images. The digitized slides are organized by case and pancreas region in an online pathology database creating a virtual biobank. Access to this online collection is currently provided to over 200 clinicians and scientists involved in type 1 diabetes research. The online database provides a means for rapid and complete data sharing and for investigators to select blocks for paraffin or frozen serial sections.

This video demonstrates procedures for characterization of human pancreatic islets using hematoxylin and eosin (H&E) and immunohistochemistry (IHC). Pancreatic sections from head, body, and tail regions are stained by both H&amp;E and IHC to determine islet endocrine composition (insulin, glucagon, and pancreatic polypeptide), cell replication (Ki67), and inflammatory infiltrates (H&E, CD3). The uncinate region is localized using IHC for pancreatic polypeptide.

인간의 췌장 Islets위한 염색법 프로토콜

Estimates of islet area and numbers and endocrine cell composition in the adult human pancreas vary from several hundred thousand to several million and ...

연구

의학

Formation of Human Prostate Epithelium Using Tissue Recombination of Rodent Urogenital Sinus Mesenchyme and Human Stem Cells

Progress in prostate cancer research is severely limited by the availability of human-derived and hormone-na&iuml;ve model systems, which limit our ability to understand genetic and molecular events underlying prostate disease initiation. Toward developing better model systems for studying human prostate carcinogenesis, we and others have taken advantage of the unique pro-prostatic inductive potential of embryonic rodent prostate stroma, termed urogenital sinus mesenchyme (UGSM). When recombined with certain pluripotent cell populations such as embryonic stem cells, UGSM induces the formation of normal human prostate epithelia in a testosterone-dependent manner. Such a human model system can be used to investigate and experimentally test the ability of candidate prostate cancer susceptibility genes at an accelerated pace compared to typical rodent transgenic studies. Since Human embryonic stem cells (hESCs) can be genetically modified in culture using inducible gene expression or siRNA knock-down vectors prior to tissue recombination, such a model facilitates testing the functional consequences of genes, or combinations of genes, which are thought to promote or prevent carcinogenesis.
The technique of isolating pure populations of UGSM cells, however, is challenging and learning often requires someone with previous expertise to personally teach. Moreover, inoculation of cell mixtures under the renal capsule of an immunocompromised host can be technically challenging. Here we outline and illustrate proper isolation of UGSM from rodent embryos and renal capsule implantation of tissue mixtures to form human prostate epithelium. Such an approach, at its current stage, requires in vivo xenografting of embryonic stem cells; future applications could potentially include in vitro gland formation or the use of induced pluripotent stem cell populations (iPSCs).

To unravel the earliest molecular mechanisms underlying prostate cancer initiation, novel and innovative human model systems and approaches are desperately needed. The potential of pre-prostatic urogenital sinus mesenchyme (UGSM) to induce pluripotent stem cell populations to form human prostate epithelium is a powerful experimental tool in prostate research.

설치류 비뇨 부비동 간엽 및 인간 줄기 세포의 조직 재조합을 사용하여 인간의 전립선 상피의 형성

Progress in prostate cancer research is severely  limited by the availability of human-derived and hormone-na&iuml;ve model systems,  which limit our ...

Preparation and Respirometric Assessment of Mitochondria Isolated from Skeletal Muscle Tissue Obtained by Percutaneous Needle Biopsy

Respirometric profiling of isolated mitochondria is commonly used to investigate electron transport chain function. We describe a method for obtaining samples of human Vastus lateralis, isolating mitochondria from minimal amounts of skeletal muscle tissue, and plate based respirometric profiling using an extracellular flux (XF) analyzer. Comparison of respirometric profiles obtained using 1.0, 2.5 and 5.0 &#956;g of mitochondria indicate that 1.0 &#956;g is sufficient to measure respiration and that 5.0 &#956;g provides most consistent results based on comparison of standard errors. Western blot analysis of isolated mitochondria for mitochondrial marker COX IV and non-mitochondrial tissue marker GAPDH indicate that there is limited non-mitochondrial contamination using this protocol. The ability to study mitochondrial respirometry in as little as 20 mg of muscle tissue allows users to utilize individual biopsies for multiple study endpoints in clinical research projects.

Methods for biopsy of Vastus lateralis, preparation of purified mitochondria, and respirometric profiling are described. The use of small muscle volume makes this technique suitable for clinical research applications.

경피적 바늘 생검에 의해 획득 골격근 조직에서 고립 된 미토콘드리아의 준비 및 호흡계 평가

Respirometric profiling of isolated mitochondria is commonly used to investigate electron transport chain function. We describe a method for obtaining ...

Application of End-to-end Anastomosis in Robotic Central Pancreatectomy

Central pancreatectomy is carried out for the treatment of benign or low-malignant potential tumors located in the pancreatic neck or proximal part of pancreatic body. With technological development, the robotic surgical system has shown its advantage in minimally invasive surgery and been increasingly applied in central pancreatectomy. However, reconstruction of the continuity of pancreas with end-to-end anastomosis after robotic central pancreatectomy has not been applied. In this study, we report surgical techniques for robotic central pancreatectomy with end-to-end anastomosis. The pancreas is reconstructed by duct-to-duct anastomosis of the pancreatic duct with a pancreatic stent inserted in the two stumps of pancreatic duct, and by end-to-end anastomosis of the pancreatic parenchyma. Compared with traditional central pancreatectomy with pancreaticoenteric anastomosis, this approach decreases the operative injury to the patient, and also conserves the integrity and continuity of the digestive duct and pancreatic duct. The robotic surgical system integrated with multiple instruments with flexible and precise movement is particularly suitable for the dissection and reconstruction of the pancreatic duct. We found that robotic central pancreatectomy with end-to-end anastomosis is safe and feasible, and we need more experience to evaluate its best indications and long-term outcomes.

The robotic central pancreatectomy with end-to-end anastomosis is feasible and safe for tumor in the pancreatic neck and proximal portion of pancreatic body. The operative techniques of this operation are presented.

로봇 중앙 Pancreatectomy 문 합 엔드 투 엔드의 응용 프로그램

Central pancreatectomy is carried out for the treatment of benign or low-malignant potential tumors located in the pancreatic neck or proximal part of ...

Immunofluorescence Labelling of Human and Murine Neutrophil Extracellular Traps in Paraffin-Embedded Tissue

Neutrophil granulocytes, also called polymorphonuclear leukocytes (PMN) due to their lobulated nucleus, are the most abundant type of leukocytes. They mature in the bone marrow and are released into the peripheral blood, where they circulate for about 6&#8722;8 h; however, in tissue, they can survive for days. By diapedesis through the endothelium, they leave the blood stream, enter tissues, and migrate towards the site of an infection following chemotactic gradients.&#160;Neutrophils can combat invading microorganisms by phagocytosis,&#160;degranulation, and generation of neutrophil extracellular traps (NETs). This protocol will help to detect NETs in paraffin-embedded tissue. NETs are the result of a process called NETosis, which leads to the release of nuclear, granular, and cytoplasmic components either from living (vital NETosis) or dying (suicidal NETosis) neutrophils. In vitro, NETs form cloud-like structures, which occupy a space several times larger than that of the cells from which they descended. The backbone of NETs is chromatin, to which a selection of proteins and peptides originating from granules and cytoplasm are bound. Thereby, a high local concentration of toxic compounds is maintained so that NETs can capture and inactivate a variety of pathogens including bacteria, fungi, viruses, and parasites, while diffusion of the highly active NET components leading to damage in neighboring tissue is limited. Nevertheless, in recent years it has become apparent that NETs, if generated in abundance or cleared insufficiently, do have pathological potential ranging from autoimmune diseases to cancer. Thus, detection of NETs in tissue samples may have diagnostic significance, and the detection of NETs in diseased tissue can influence the treatment of patients. Since paraffin-embedded tissue samples are the standard specimen used for pathological analysis, it was sought to establish a protocol for fluorescent staining of NET components in paraffin-embedded tissue using commercially available antibodies.

Neutrophil extracellular traps (NETs) are three-dimensional structures generated by stimulated neutrophil granulocytes. It has become clear in recent years that NETs are involved in a wide variety of diseases. Detection of NETs in tissue may have diagnostic relevance, so standardized protocols for labelling NET components are required.

파라핀 내장 된 조직에서 인간과 뮤린 호중구 세포 외 트랩의 면역 형광 라벨링

Neutrophil granulocytes, also called polymorphonuclear leukocytes (PMN) due to their lobulated nucleus, are the most abundant type of leukocytes. They ...

Robotic Enucleation of an Intra-Pancreatic Insulinoma in the Pancreatic Head

Pancreatic parenchyma sparing surgery for insulinomas avoids the risk of endocrine and exocrine insufficiency, and potential high-risk anastomoses associated with pancreatic resection. Robotic surgery may be used as an alternative for open pancreatic enucleation without compromising dexterity and 3D-vision.
We present the case of a 42-year old woman who presented with sweating, tremor and episodes of hypoglycemia. A fasting test confirmed endogenic insulin overproduction. After inconclusive CT- and MRI imaging, endoscopic ultrasonography showed a hypoechoic lesion, which was fully within the pancreatic head. Although consent was obtained for pancreatoduodenectomy, robotic enucleation seemed feasible. After mobilization, intraoperative ultrasonography was used to identify the lesion and its relation with the pancreatic duct. Dissection was performed using a traction suture, hot shears and bipolar diathermia. A sealant patch was applied for hemostasis and a drain placed. The patient developed a grade B pancreatic fistula for which endoscopic sphincterotomy was performed; the surgical drain could be removed in the outpatient clinic after 20 days. Prospective studies should confirm the short- and long-term benefits of robotic enucleation of insulinomas.

Here, we present a robotic approach to enucleate an insulinoma in the pancreatic head.

췌장 머리에 있는 췌장 내 인슐린종의 로봇 핵형성

Pancreatic parenchyma sparing surgery for insulinomas avoids the risk of endocrine and exocrine insufficiency, and potential high-risk anastomoses ...

Measurement of Tissue Non-Heme Iron Content using a Bathophenanthroline-Based Colorimetric Assay

Iron is an essential micronutrient. Both iron overload and deficiency are highly detrimental to humans, and tissue iron levels are finely regulated. The use of experimental animal models of iron overload or deficiency has been instrumental to advance knowledge of the mechanisms involved in the systemic and cellular regulation of iron homeostasis. The measurement of total iron levels in animal tissues is commonly performed with atomic absorption spectroscopy or with a colorimetric assay based on the reaction of non-heme iron with a bathophenanthroline reagent. For many years, the colorimetric assay has been used for the measurement of the non-heme iron content in a wide range of animal tissues. Unlike atomic absorption spectroscopy, it excludes the contribution of heme iron derived from hemoglobin contained in red blood cells. Moreover, it does not require sophisticated analytical skills or highly expensive equipment, and can thus be easily implemented in most laboratories. Finally, the colorimetric assay can be either cuvette-based or adapted to a microplate format, allowing higher sample throughput. The present work provides a well-established protocol that is suited for the detection of alterations in tissue iron levels in a variety of experimental animal models of iron overload or iron deficiency.

Here, a protocol for the measurement of the non-heme iron content in animal tissues is provided, using a simple, well-established colorimetric assay that can be easily implemented in most laboratories.

Bathophenanthroline-based Colorimetric Assay를 이용한 조직 비헴철 함량 측정

Iron is an essential micronutrient. Both iron overload and deficiency are highly detrimental to humans, and tissue iron levels are finely regulated. The ...

Oral Submucous Fibrosis Tissue Sample Preparation for Hematoxylin-Eosin Staining

Hematoxylin-Eosin 염색을 위한 구강 점막하 섬유증 조직 샘플 준비

Tissue Embedding and Immunofluorescence Staining of Myocardial Sleeves in Murine Pulmonary Veins

<meta charset="utf8"></head><body>설치류 안구의 망막 두께와 신경절 세포층 수 평가

Preparation and Analysis of Histological Slides of Rat and Mouse Eyeballs to Evaluate the Retina

A rodent eyeball is a powerful tool for researching the pathomechanisms of many ophthalmic diseases, such as glaucoma, hypertensive retinopathy, and many more. Preclinical experiments enable researchers to examine the efficacy of novel drugs, develop new methods of treatment, or seek new pathomechanisms involved in the disease's onset or progression. A histological examination provides a lot of information necessary to assess the effects of the conducted experiments and can reveal degeneration, tissue remodeling, infiltration, and many other pathologies. In clinical research, there is rarely any chance of obtaining eye tissue suitable for a histological examination, which is why researchers should take advantage of the opportunity offered by the examination of eyeballs from rodents. This manuscript presents a protocol for the histological preparation of rodent eyeballs' sections. The procedure is presented for the eyeballs of mice and rats and has the following steps: (i) harvesting the eyeball, (ii) preserving the eyeball for further analysis, (iii) processing the tissue in paraffin, (iv) preparing slides, (v) staining with hematoxylin and eosin, (vi) assessing the tissue under a light microscope. With the proposed method, the retina can be easily visualized and assessed in detail.

<meta charset="utf8"></head><body>망막을 평가하기 위한 쥐와 쥐 안구의 조직학적 슬라이드의 준비 및 분석

This manuscript presents a protocol for the histological preparation of slides from rodent eyeballs. With the proposed method, the inner retina can be easily visualized and assessed under a light microscope. The procedure is presented for the eyeballs of mice and rats.

망막을 평가하기 위한 쥐와 쥐 안구의 조직학적 슬라이드의 준비 및 분석

A rodent eyeball is a powerful tool for researching the pathomechanisms of many ophthalmic diseases, such as glaucoma, hypertensive retinopathy, and many ...