Sophia Ackermann, Maximilian Herold, Vincent Rohrbacher, Michael Schäfer, Marcell Tóth, Stefan Thomann, Thilo Hackert, Eduard Ryschich
{"title":"利用光纤显微镜对重要的人体胰腺组织进行原位免疫荧光成像。","authors":"Sophia Ackermann, Maximilian Herold, Vincent Rohrbacher, Michael Schäfer, Marcell Tóth, Stefan Thomann, Thilo Hackert, Eduard Ryschich","doi":"10.1155/2024/1397875","DOIUrl":null,"url":null,"abstract":"<p><strong>Purpose: </strong>Surgical resection is the only curative option for pancreatic carcinoma, but disease-free and overall survival times after surgery are limited due to early tumor recurrence, most often originating from local microscopic tumor residues (R1 resection). The intraoperative identification of microscopic tumor residues within the resection margin <i>in situ</i> could improve surgical performance. The aim of this study was to evaluate the effectiveness of fiber-optic microscopy for detecting microscopic residues in vital pancreatic cancer tissues. <i>Experimental Design</i>. Fresh whole-mount human pancreatic tissues, histological tissue slides, cell culture, and chorioallantoic membrane xenografts were analyzed. Specimens were stained with selected fluorophore-conjugated antibodies and studied using conventional wide-field and self-designed multicolor fiber-optic fluorescence microscopy instruments.</p><p><strong>Results: </strong>Whole-mount vital human tissues and xenografts were stained and imaged using an <i>in situ</i> immunofluorescence protocol. Fiber-optic microscopy enabled the detection of epitope-based fluorescence in vital whole-mount tissue using fluorophore-conjugated antibodies and enabled visualization of microvascular, epithelial, and malignant tumor cells. Among the selected antigen-antibody pairs, antibody clones WM59, AY13, and 9C4 were the most promising for fiber-optic imaging in human tissue samples and for endothelial, tumor and epithelial cell detection.</p><p><strong>Conclusions: </strong>Fresh dissected whole-mount tissue can be stained using direct exposure to selected antibody clones. Several antibody clones were identified that provided excellent immunofluorescence imaging of labeled structures, such as endothelial, epithelial, or EGFR-expressing cells. The combination of <i>in situ</i> immunofluorescence staining and fiber-optic microscopy visualizes structures in vital tissues and could be proposed as an useful tool for the <i>in situ</i> identification of residual tumor mass in patients with a high operative risk for incomplete resection.</p>","PeriodicalId":47063,"journal":{"name":"International Journal of Biomedical Imaging","volume":"2024 ","pages":"1397875"},"PeriodicalIF":3.3000,"publicationDate":"2024-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11178408/pdf/","citationCount":"0","resultStr":"{\"title\":\"<i>In Situ</i> Immunofluorescence Imaging of Vital Human Pancreatic Tissue Using Fiber-Optic Microscopy.\",\"authors\":\"Sophia Ackermann, Maximilian Herold, Vincent Rohrbacher, Michael Schäfer, Marcell Tóth, Stefan Thomann, Thilo Hackert, Eduard Ryschich\",\"doi\":\"10.1155/2024/1397875\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Purpose: </strong>Surgical resection is the only curative option for pancreatic carcinoma, but disease-free and overall survival times after surgery are limited due to early tumor recurrence, most often originating from local microscopic tumor residues (R1 resection). The intraoperative identification of microscopic tumor residues within the resection margin <i>in situ</i> could improve surgical performance. The aim of this study was to evaluate the effectiveness of fiber-optic microscopy for detecting microscopic residues in vital pancreatic cancer tissues. <i>Experimental Design</i>. Fresh whole-mount human pancreatic tissues, histological tissue slides, cell culture, and chorioallantoic membrane xenografts were analyzed. Specimens were stained with selected fluorophore-conjugated antibodies and studied using conventional wide-field and self-designed multicolor fiber-optic fluorescence microscopy instruments.</p><p><strong>Results: </strong>Whole-mount vital human tissues and xenografts were stained and imaged using an <i>in situ</i> immunofluorescence protocol. Fiber-optic microscopy enabled the detection of epitope-based fluorescence in vital whole-mount tissue using fluorophore-conjugated antibodies and enabled visualization of microvascular, epithelial, and malignant tumor cells. Among the selected antigen-antibody pairs, antibody clones WM59, AY13, and 9C4 were the most promising for fiber-optic imaging in human tissue samples and for endothelial, tumor and epithelial cell detection.</p><p><strong>Conclusions: </strong>Fresh dissected whole-mount tissue can be stained using direct exposure to selected antibody clones. Several antibody clones were identified that provided excellent immunofluorescence imaging of labeled structures, such as endothelial, epithelial, or EGFR-expressing cells. The combination of <i>in situ</i> immunofluorescence staining and fiber-optic microscopy visualizes structures in vital tissues and could be proposed as an useful tool for the <i>in situ</i> identification of residual tumor mass in patients with a high operative risk for incomplete resection.</p>\",\"PeriodicalId\":47063,\"journal\":{\"name\":\"International Journal of Biomedical Imaging\",\"volume\":\"2024 \",\"pages\":\"1397875\"},\"PeriodicalIF\":3.3000,\"publicationDate\":\"2024-06-06\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11178408/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"International Journal of Biomedical Imaging\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1155/2024/1397875\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2024/1/1 0:00:00\",\"PubModel\":\"eCollection\",\"JCR\":\"Q2\",\"JCRName\":\"ENGINEERING, BIOMEDICAL\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"International Journal of Biomedical Imaging","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1155/2024/1397875","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/1/1 0:00:00","PubModel":"eCollection","JCR":"Q2","JCRName":"ENGINEERING, BIOMEDICAL","Score":null,"Total":0}
In Situ Immunofluorescence Imaging of Vital Human Pancreatic Tissue Using Fiber-Optic Microscopy.
Purpose: Surgical resection is the only curative option for pancreatic carcinoma, but disease-free and overall survival times after surgery are limited due to early tumor recurrence, most often originating from local microscopic tumor residues (R1 resection). The intraoperative identification of microscopic tumor residues within the resection margin in situ could improve surgical performance. The aim of this study was to evaluate the effectiveness of fiber-optic microscopy for detecting microscopic residues in vital pancreatic cancer tissues. Experimental Design. Fresh whole-mount human pancreatic tissues, histological tissue slides, cell culture, and chorioallantoic membrane xenografts were analyzed. Specimens were stained with selected fluorophore-conjugated antibodies and studied using conventional wide-field and self-designed multicolor fiber-optic fluorescence microscopy instruments.
Results: Whole-mount vital human tissues and xenografts were stained and imaged using an in situ immunofluorescence protocol. Fiber-optic microscopy enabled the detection of epitope-based fluorescence in vital whole-mount tissue using fluorophore-conjugated antibodies and enabled visualization of microvascular, epithelial, and malignant tumor cells. Among the selected antigen-antibody pairs, antibody clones WM59, AY13, and 9C4 were the most promising for fiber-optic imaging in human tissue samples and for endothelial, tumor and epithelial cell detection.
Conclusions: Fresh dissected whole-mount tissue can be stained using direct exposure to selected antibody clones. Several antibody clones were identified that provided excellent immunofluorescence imaging of labeled structures, such as endothelial, epithelial, or EGFR-expressing cells. The combination of in situ immunofluorescence staining and fiber-optic microscopy visualizes structures in vital tissues and could be proposed as an useful tool for the in situ identification of residual tumor mass in patients with a high operative risk for incomplete resection.
期刊介绍:
The International Journal of Biomedical Imaging is managed by a board of editors comprising internationally renowned active researchers. The journal is freely accessible online and also offered for purchase in print format. It employs a web-based review system to ensure swift turnaround times while maintaining high standards. In addition to regular issues, special issues are organized by guest editors. The subject areas covered include (but are not limited to):
Digital radiography and tomosynthesis
X-ray computed tomography (CT)
Magnetic resonance imaging (MRI)
Single photon emission computed tomography (SPECT)
Positron emission tomography (PET)
Ultrasound imaging
Diffuse optical tomography, coherence, fluorescence, bioluminescence tomography, impedance tomography
Neutron imaging for biomedical applications
Magnetic and optical spectroscopy, and optical biopsy
Optical, electron, scanning tunneling/atomic force microscopy
Small animal imaging
Functional, cellular, and molecular imaging
Imaging assays for screening and molecular analysis
Microarray image analysis and bioinformatics
Emerging biomedical imaging techniques
Imaging modality fusion
Biomedical imaging instrumentation
Biomedical image processing, pattern recognition, and analysis
Biomedical image visualization, compression, transmission, and storage
Imaging and modeling related to systems biology and systems biomedicine
Applied mathematics, applied physics, and chemistry related to biomedical imaging
Grid-enabling technology for biomedical imaging and informatics