Abstract:Temporal and spatial regulation of proteins contributes to function. We describe a multidimensional microscopic robot technology for high-throughput protein colocalization studies that runs cycles of fluorescence tagging, imaging and bleaching in situ. This technology combines three advances: a fluorescence technique capable of mapping hundreds of different proteins in one tissue section or cell sample; a method selecting the most prominent combinatorial molecular patterns by representing the data as binary ve… Show more
“…9). Extensive literature exists on the development of multiplexed and more recently hyperplexed fluorescence methods for tissue analyses, including instrumentation, software, and bioinformatics tools that can be used in the development of diagnostic tests 9, 20, 21, 22, 23. The ability to measure many parameters in large numbers of tissue samples has propelled these assays into the realm of big data challenges for the future.…”
Section: Hyperplexed Fluorescence Measurements Of the Cellular And Sumentioning
The high‐content interrogation of single cells with platforms optimized for the multiparameter characterization of cells in liquid and solid biopsy samples can enable characterization of heterogeneous populations of cells ex vivo. Doing so will advance the diagnosis, prognosis, and treatment of cancer and other diseases. However, it is important to understand the unique issues in resolving heterogeneity and variability at the single cell level before navigating the validation and regulatory requirements in order for these technologies to impact patient care. Since 2013, leading experts representing industry, academia, and government have been brought together as part of the Foundation for the National Institutes of Health (FNIH) Biomarkers Consortium to foster the potential of high‐content data integration for clinical translation.
“…9). Extensive literature exists on the development of multiplexed and more recently hyperplexed fluorescence methods for tissue analyses, including instrumentation, software, and bioinformatics tools that can be used in the development of diagnostic tests 9, 20, 21, 22, 23. The ability to measure many parameters in large numbers of tissue samples has propelled these assays into the realm of big data challenges for the future.…”
Section: Hyperplexed Fluorescence Measurements Of the Cellular And Sumentioning
The high‐content interrogation of single cells with platforms optimized for the multiparameter characterization of cells in liquid and solid biopsy samples can enable characterization of heterogeneous populations of cells ex vivo. Doing so will advance the diagnosis, prognosis, and treatment of cancer and other diseases. However, it is important to understand the unique issues in resolving heterogeneity and variability at the single cell level before navigating the validation and regulatory requirements in order for these technologies to impact patient care. Since 2013, leading experts representing industry, academia, and government have been brought together as part of the Foundation for the National Institutes of Health (FNIH) Biomarkers Consortium to foster the potential of high‐content data integration for clinical translation.
“…Described previously, the multi-epitope-ligand cartography (MELC) 8,9 , is one of these bleaching techniques, that is capable to map the location of different proteins in one sample of cells or tissues using sequential rounds of fluorescent detection 10 . In each cycle, a couple of antibodies are added; phase contrast and fluorescence images then are acquired by a high-sensitivity "charged Coupled Device", a sensor used in digital cameras to record images; the sample is washed with phosphate buffer saline and bleaches at the excitation wavelengths; and post bleaching phase contrast and fluorescence images are acquired.…”
Multiplexed immunofluorescence (IF) methods to detect simultaneously different molecules are revolutionizing immunohistochemistry (IHC) in the last years. These new technologies can be valuable for tumor examination in formalin-fixed paraffin-embedded (FFPE) specimens, and for improved new treatment discoveries and translational cancer studies. The aim of this minireview is to highlight the recent methodologies that using multiplexed IF to study simultaneous proteins identification in FFPE tumor tissues to clinical research and potential translational analysis. Multiplexed IF methods, which permit the identification of up to 4 proteins at the same time, have been increased in the last years the abilities of study cells by cells and their spatial distribution in several tumor tissues. Although, most of the old platforms are not more used after the powerful multiplex IHC methods are continue growing, the basis of these old methodologies have helped to improve the new technologies. Associated with image analysis software's these technologies can be improved to performance high throughput assay to study these specimens. Each multiplexed IF technique, detailed herein, is associated with important advantages in cancer study as well as translational research studies.
“…This can be exploited to develop new diagnostic methods. Where single genetic markers are not sufficient to distinguish healthy tissue from a diseased phenotype, combinations of genes can enhance diagnostic strength [5]. Microscopy and comparative analysis of images is therefore becoming an important tool to study the spatio-temporal distribution of multiple gene products.…”
Abstract. Carefully studied in-situ hybridization Gene expression patterns (GEP) can provide a first glance at possible relationships among genes. Automatic comparative analysis tools are an indispensable requirement to manage the constantly growing amount of such GEP images. We present here an automated processing pipeline for Segmenting, Classification, and Clustering large-scale sets of Drosophila melanogaster GEP images that facilitates automatic GEP analysis.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.