The Translational Pathology Core Laboratory (TPCL) is a research facility in the UCLA Department of Pathology and Laboratory Medicine and a UCLA Jonsson Comprehensive Cancer Center Shared Facility. Since 1996, the TPCL has provided an array of pathology-related services in support of basic, translational and clinical research at UCLA. Our mission is to provide the highest quality pathology services in an efficient and cost-effective manner. The TPCL also provides expert consultative services to investigators in pathology-related study design, tissue selection, microscopic interpretation, immunohistochemistry, laser capture microdissection, digital image analysis, and IRB-related tissue questions.
Digital imaging and image analysis services include state-of-the-art virtual microscopy (VM) and digital pathology (DP) (image analysis) services to the UCLA community. In VM, whole glass slides are converted to high resolution digital images (either brightfield or fluorescence) for easy archiving and retrieval, detailed magnification up to 40X, remote viewing via a web-based interface and preparation for publications and teaching - most of which can be performed from a single computer, using free software. DP includes performance of (or instruction in performing) quantitative digital image analysis studies. This includes both quantitative immunohistochemistry and analysis of other cellular characteristics (e.g., cell size). Dr. Clara Magyar directs the digital imaging/image analysis services for the core. The TPCL provides assistance with image acquisition and analysis, tips on sample preparation, and training on our image analysis systems. The TPCL houses several different scanners and image analysis programs (discussed in detail below). Scanners include the Applied Imaging Leica Aperio Versa high throughput scanning system (fluorescence, brightfield) and the Aperio ScanScope AT high throughput scanning system (brightfield, web enabled).Our automated digital analysis software includes Definiens, Ariol and Molecular Devices MetaMorph.
Why use digital imaging/image analysis? VM/DP permits real-time discussions of histology images posted on secure websites, eliminating barriers to the exchange of pathology information with national/international collaborators/consultants, helping to reduce inter/intra-observer variability that adversely affects quantitative histopathological analysis, improving the overall efficiency of our researchers and clinicians, and eliminating delay, damage, or loss of often irreplaceable slides during shipping between UCLA faculty and extramural sites. Images can be archived, stored indefinitely, and easily retrieved - unlike glass slides, which may be lost, misfiled, damaged or fade with time.
The TPCL houses several different scanners and image analysis programs (discussed in detail below).Scanners include the Leica Aperio Versa high throughput scanning system (fluorescence) and the Aperio ScanScope AT high throughput scanning system (brightfield, web enabled). For image analysis, we use Definiens Tissue Studio.
Aperio ScanScope AT. The ScanScope AT is an automated scanning system that utilizes Aperio's linear-array scanning technology to produce high quality images of whole slides at either 20x (standard) or 40x. Using the ImageScope software, users can view images on a continuum of magnifications ranging from 1x to the maximum scanned magnification. Additionally, users can make annotations on their images and export TIFF and JPEG snapshots from the field of view. The software permits simultaneous display of multiple scanned slides. Multiple images can also be "locked" so that all slides move synchronously, which allows users to follow same region from slide to slide. Scanned slide images are saved to a local server DVD, or external hard drive along with a copy of the ImageScope software. The ScanScope AT system is ideal for brightfield imaging and has been utilized at the TPCL for research, publications, teaching (Department of Pathology and Laboratory Medicine and David Geffen School of Medicine at UCLA), and archiving of slides.
Leica Aperio Versa. Versa is an automated scanning microscope. With Leica Aperio Versa, we are able to scan fluorescent slides. For immunofluorescent staining, Versa is capable of detecting up to 4 probes/fluorochromes.
Definiens. Definiens offers the world's most advanced technology for extracting intelligence from images. Instead of examining a given image pixel by pixel, Definiens' segmentation and classification processes recognize groups of pixels as objects, picking out shapes,colors and textures. The technology examines objects in relationship to each other, and can understand scale, overlapping objects and the relationship of two-dimensional images to three-dimensional shapes. Using these factors, the technology identifies individual objects within an image and makes precise and detailed measurements. It can handle the inherent variability in living organisms and can even recognize anomalies, such as an abnormal tumor cell. Definiens' technology will enable us to analyze vast numbers of images quickly and accurately. Definiens also has the capability of analyzing entire images from many different platforms and formats thereby allowing the users of the TPCL a greater deal of versatility in the type of images that may be analyzed.
Rules for slides to be scanned:
Aperio: Best results are obtained with glass slides with tissues of regular thickness (4-5 microns), and coverslipped. Slides with marks (such as ink dots), extra mounting medium, non-glass slides, plastic cover slips, thick labels and labels that are going around the slide present challenges for proper imaging, and may need to be removed in order to obtain an optimal scan. Images are individually reviewed once the scanning is completed. Scans with a poor quality will be rescanned manually to achieve the maximum quality possible. Faint stains, air bubbles, thick sections, or marks on the glass surface can cause lower quality scans. In some cases, we recommend sample recut or restaining.
Versa: Best results are obtained with glass slides with tissues of regular thickness (4-5 microns). Slides must be coverslipped. Coverslips cannot be mounted anywhere on the slide label region. The slide label must be completely dry and absent of any mounting residue. Avoid sample placement near slide edges as samples may be beyond the physical scanning limits of the microscope.
Histology-related services include fixation, processing, embedding, and sectioning of human and animal tissues, cryostat sections (frozen tissues), sectioning for RT-PCR or other molecular studies, H&E staining and decalcification of blocks. Special stains, as performed in our diagnostic histology laboratory, are available as well.
Services include immunostaining of established antibodies as well as optimization of new antibodies and double staining for brightfield; TUNEL assay for apoptosis is also performed. We have a large library of established antibodies available for use on human and animal tissues [IHC Core Antibodies]. Several chromogens (DAB, Fast Red, AP Blue, AP Green) and counterstains (hematoxylin, methyl-green, nuclear fast red) are available. The detection system most frequently used is the HP system using DAB with hematoxylin counterstain.
In 2010, TPCL and the California NanoSystems Institutes Advanced Light Microscopy/Spectroscopy (ALMS) core lab jointly purchased a Leica Laser-Microdissection (LMD) 7000. The TPCL also houses an Arcturus PixCell IIe with single-cell and flourescence capabilities.
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