Advancing Biomedical Research: The Flow Cytometry and Advanced Technologies at the Medical College of Wisconsin
The Medical College of Wisconsin (MCW) stands at the forefront of biomedical research, offering a comprehensive suite of advanced technologies and shared resources designed to propel scientific discovery. Among these, the Flow Cytometry Shared Resource (FLOW Core) plays a pivotal role, providing essential tools and expertise for analyzing cells at a granular level. However, the FLOW Core is part of a much larger ecosystem of cutting-edge facilities that collectively empower researchers across MCW and its affiliated institutions. This article delves into the diverse array of these resources, highlighting their capabilities and their collective impact on advancing our understanding of health and disease.
The Heart of Cellular Analysis: Flow Cytometry at MCW
The Flow Cytometry Shared Resource (FLOW Core) at MCW, supported by the Children’s Research Institute (CRI) and the Medical College of Wisconsin Cancer Center (MCWCC), is a critical component of the institution's research infrastructure. Its mission is to provide investigators with comprehensive support for multicolor analytical flow cytometry and cell sorting. This includes crucial technical assistance and training, operator-assisted cell sorting in both standard and BSL-2+ modes, and expert consultation for application development, assay design, and data analysis. Furthermore, the FLOW Core offers advanced data interpretation, ensuring that researchers can derive meaningful insights from their experiments.
The facility is equipped with state-of-the-art instrumentation, including two Cytek Aurora spectral cytometers, an LSRFortessa X20 analytical cytometer, a NanoCellect WOLF cell sorter, a Miltenyi MACSQuant Tyto cell sorter, and a BD FACSAriaII cell sorter. Complementing these are a HemaVet 950 CBC Analyzer for complete blood counts and a DeNovix CellDrop FL cell counter for accurate cell counting and viability assessment.
The impact of flow cytometry at MCW is substantial. The Flow Cytometry Laboratory at the Medical College of Wisconsin/Wisconsin Diagnostic Laboratories, directed by Dr. [Director's Name - information not provided], is a busy service, performing over 4200 studies annually. This robust activity underscores its importance in supporting a growing, high-volume patient population, particularly those with leukemia, lymphoma, and those undergoing bone marrow transplantation. The hematopathology team leverages sophisticated flow cytometry studies for the diagnosis of acute leukemia, myelodysplastic syndrome, plasma cell myeloma, non-Hodgkin lymphoma, and paroxysmal nocturnal hemoglobinuria. Their expertise extends to the management of these disorders, including sensitive minimal residual disease assessment. The laboratory currently performs 8-color flow cytometry and is actively transitioning to a 10-color system, further enhancing its analytical capabilities.
Research utilizing flow cytometry is published and cited extensively. For instance, a publication on "Flow Cytometry of B-Cell Neoplasms" by Kroft SH and Harrington AM, published in 2017, highlights the standard role of flow cytometric evaluation in diagnosing most B-cell lymphoproliferative disorders. The abstract emphasizes that establishing a neoplastic B-cell population relies on identifying light chain restriction or lack of expression in mature neoplasms, alongside demonstrating aberrant antigen expression in both immature and mature neoplasms compared to their normal counterparts. The authors detail the immunophenotypes of common B-cell neoplasms, focusing on differential diagnosis and prognostic and therapeutic implications. This work, with 21 citations, underscores the foundational importance of flow cytometry in hematological diagnostics and research.
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Beyond Flow Cytometry: A Multitude of Advanced Research Cores
MCW's commitment to fostering cutting-edge research extends far beyond flow cytometry, encompassing a broad spectrum of specialized cores and shared resources. These facilities provide access to advanced instruments and technologies that might otherwise be inaccessible to individual faculty members. Expert leaders and dedicated staff offer comprehensive laboratory and data analysis services, alongside invaluable training for faculty, staff, and trainees, all in support of investigator-initiated studies. The professional services provided span experimental consultation, execution, and meticulous documentation, aiding scientific projects from their conceptualization through to publication. The overarching mission of the MCW Core Facilities is to support all MCW and MCW-affiliated investigators and physician scientists, ensuring they have the resources necessary to pursue groundbreaking research.
Imaging and Microscopy for Unveiling Cellular Architecture
The realm of microscopy and imaging is exceptionally well-represented at MCW. The Children’s Research Institute (CRI) Histology Core, a CAP-accredited laboratory, offers a full spectrum of routine and specialized histological and immunological services. This core is staffed by ASCP-certified Histologists and benefits from the analytical expertise of an American Board of Pathology-certified Pathologist, ensuring high-quality diagnostic and research support.
Further enhancing microscopic capabilities, CRI's imaging core houses a variety of state-of-the-art microscopic imaging systems. These include several confocal microscopes, a laser microdissection system for precise isolation of cellular components, and whole slide scanners for comprehensive tissue analysis. This imaging core is accessible to all investigators, including those at collaborating institutes, fostering inter-institutional research collaborations.
The OxCAM-EM facility integrates both Light and Electron Microscopy services, open to all MCW investigators, affiliated institutions, and external researchers. The Electron Microscopy laboratory provides comprehensive services such as tissue processing, immunoelectron microscopy, negative staining, enzyme cytochemistry, ultrastructural electron tomography, and essential TEM training.
The Neuroscience Research Center's microscopy core is specifically equipped to support specialized imaging of neuronal tissue. It features a variety of confocal imaging systems, notably the AxL Cleared Tissue Light Sheet Microscope. This advanced system is complemented by sophisticated software packages, including the Bitplane Imaris 10.0 F1 package and SVI Huygens Professional deconvolution software, enabling detailed analysis and visualization of complex neural structures.
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For structural biology investigations, the facility includes an X-ray diffraction system. This comprises a Rigaku R-AXIS IV++ image plate detector system, a MicroMax 007 generator, and an automated crystallization system equipped with a CrysCam for nanoliter crystallization and visualization, crucial for determining the three-dimensional structures of biomolecules.
The Biomedical Imaging Shared Resource (BISR) plays a vital role in advancing research at MCW by providing both expertise and cutting-edge technology for preclinical imaging. Their capabilities include bioluminescence and fluorescence imaging, low-dose X-ray microCT, and Cherenkov Luminescence imaging. Furthermore, BISR offers preclinical near-infrared (NIR) and short-wave infrared (SWIR) imaging, significantly enhancing the translational potential of research in both non-cancer and cancer-related fields.
Omics and Genomics: Decoding the Blueprint of Life
MCW provides extensive resources for researchers exploring the complexities of the genome, transcriptome, epigenome, and proteome. The Mellowes Center is a hub for advanced sequencing technologies, offering both short-read (Illumina) and long-read (Oxford Nanopore) sequencing. It also supports single-cell sequencing and spatial genomic analysis, as well as NanoString nCounter applications. This comprehensive suite of services allows for in-depth investigation across multiple molecular layers.
The Mellowes Center Bioinformatics Resource is an integral part of this offering, providing essential assistance in data processing and offering services or collaborative research support to MCW investigators. Their analysis services encompass crucial aspects of the research lifecycle, including study design, data acquisition, rigorous data analysis, and support for publication.
Protein Production and Structural Biology: Illuminating Molecular Mechanisms
The Structural Biology Shared Resources (SBSR) facility is a two-component entity crucial for understanding the molecular underpinnings of biological processes. Its protein production facility is dedicated to supplying high-quality, homogeneous proteins for structural biology and other applications. This is achieved through the utilization of a variety of expression systems, including baculovirus/insect cell, HEK293, CHO, and E.coli, ensuring researchers have access to the specific proteins they need for their studies.
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Biochemistry and Mass Spectrometry: Quantifying and Characterizing Molecules
The Biochemistry Mass Spectrometry shared equipment houses a Thermo LTQ Mass Spectrometer, readily available for walk-up use for peptide and protein analysis. This resource is accessible to faculty, students, staff within the Department of Biochemistry, and their collaborators, facilitating detailed molecular characterization.
Advanced Spectroscopy and Biomolecular Analysis
The NMR Facility, an interdepartmental research service unit within the Biochemistry Department, offers high-field NMR spectroscopy. This powerful technique is indispensable for the study of biomolecular structure and dynamics, providing detailed insights into the arrangement and movement of molecules within biological systems.
Analytical biochemical studies and the investigation of biomolecular structure and dynamics, from proteins to small molecules, are vital to understanding the molecular interactions that influence health and disease.
Redox Biology and Metabolic Insights
The Redox and Bioenergetics Shared Resource (RBSR) supports critical metabolic services. These include the measurement of reactive oxygen species (ROS), metabolic pathway profiling, oxygen consumption analysis, assessment of glycolytic function, membrane potential determination, and the study of metabolic reprogramming. These analyses are performed using advanced instrumentation such as Agilent Seahorse, HPLC, UHPLC-electron paramagnetic resonance (EPR), and luminescence spectrometers, providing a detailed understanding of cellular energy metabolism and oxidative stress.
Cellular Function and Long-Term Imaging
The NMR Facility also supports automated image acquisition, allowing for the analysis of incubated cells over extended periods-days, weeks, or even months. This capability is crucial for studying cellular behavior and responses in a stable, maintained environment over time.
Stem Cell Research and Translational Applications
The Cardiovascular iPSC Core provides cutting-edge induced pluripotent stem cell (iPSC) services to research teams across MCW. This core facilitates translational research by bridging preclinical animal and cell models with human cells and tissues. Their services encompass human iPSC reprogramming from peripheral blood monocytes or fibroblasts (skin biopsy), differentiation of human iPSC lines into endothelial cells, cardiomyocytes, or vascular smooth muscle cells, and iPSC CRISPR gene knockout/knock-in or single nucleotide editing. Additionally, the core offers premade cell products from their bank or select validated external sources, accelerating research timelines.
Tissue Banking for Comprehensive Biospecimen Services
The CAP-accredited MCW Tissue Bank provides essential services related to human biospecimens for MCW faculty investigators. Beyond the distribution of tissues and other biospecimens, their services include tissue and blood processing, RNA/DNA extraction, tissue analytics, and secure storage, ensuring the integrity and availability of critical research materials.
Clinical Trials and Translational Support
The Department of Radiology, in collaboration with the Medical College of Wisconsin’s Clinical Cancer Center (MCWCC) Clinical Trials Office, actively supports the assessment of tumor response to therapy. This is guided by board-certified radiologists, integrating imaging expertise with clinical research.
Data Science and Computational Analysis
MCW's data science facilities are robust, encompassing geospatial analysis, epidemiology, biostatistics, bioinformatics, and qualitative analysis. These are achieved through the application of advanced computational techniques, enabling researchers to extract insights from complex datasets.
Preclinical Studies and Translational Resources
Basic and translational science investigators are well-served by advanced technologies covering cellular function, tissue analytics, genetic mechanisms, biophysics, and molecular analysis. MCW Core Facilities and Shared Resources offer high-quality services for complex preclinical studies, available for both whole animal and in vitro applications using established and novel research models. MCW's translational resources provide clinical-grade services adapted to investigator protocols for research involving adult and pediatric participants and model systems.
Biomechanics and Injury Research
The Clement J. Zablocki VA Medical Center Laboratories house a wide range of facilities that support research into the biomechanics of injury, further broadening the scope of research capabilities available through MCW affiliations.
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