Digital Transplantation Pathology: Combining Whole Slide Imaging, Multiplex Staining and Automated Image Analysis

Abstract

Conventional histopathology is the gold standard for allograft monitoring, but its value proposition is increasingly questioned. “-Omics” analysis of tissues, peripheral blood and fluids and targeted serologic studies provide mechanistic insights into allograft injury not currently provided by conventional histology. Microscopic biopsy analysis, however, provides valuable and unique information: a) spatial-temporal relationships; b) rare events/cells; c) complex structural context; and d) integration into a “systems” model. Nevertheless, except for immunostaining, no transformative advancements have “modernized” routine microscopy in over 100 years. Pathologists now team with hardware and software engineers to exploit remarkable developments in digital imaging, nanoparticle multiplex staining, and computational image analysis software to bridge the traditional histology - global “–omic” analyses gap. Included are side-by-side comparisons, objective biopsy finding quantification, multiplexing, automated image analysis, and electronic data and resource sharing. Current utilization for teaching, quality assurance, conferencing, consultations, research and clinical trials is evolving toward implementation for low-volume, high-complexity clinical services like transplantation pathology. Cost, complexities of implementation, fluid/evolving standards, and unsettled medical/legal and regulatory issues remain as challenges. Regardless, challenges will be overcome and these technologies will enable transplant pathologists to increase information extraction from tissue specimens and contribute to cross-platform biomarker discovery for improved outcomes.

Automated recognition and quantification of pancreatic islets in Zucker diabetic fatty rats treated with exendin-4

“…Development of a digital scanner has enabled whole slide access with high resolution and provided more tissues to be analyzed. Most of the analytic software is pixel-based, one that isolates and quantifies colored pixels such as immunopositive areas (Rojo et al 2009, Prasad & Prabhu 2011, Isse et al 2012. Segregation of specific target tissues is still particularly challenging, such as in the case of ZDF islets (Figs 2, 3 and 4) with a morphologically intricate structure.…”

“…In the case of normal islets with homogeneous insulin immunostaining, the value of insulin-stained area that can be quantified easily using automatic isolation of colored pixels (Rojo et al 2009, Prasad & Prabhu 2011, Isse et al 2012) is very similar to that of the islet area. However, the area of ZDF islets with depleted insulin immunostaining cannot be quantified by the colored pixel-based measurement.…”

Novel Platforms of Multiplexed Immunofluorescence for Study of Paraffin Tumor Tissues

“…Although, multiplexed IHC staining available for FFPE material enables multi-parametric readouts from a single tissue section, the different techniques described before, have some tines limited scalability and throughput, related to limited small region-of-interests (ROI) scanning or limited to few number of fields-of-views 41,42 as summarized in the Table 2. The major part of the scanner system ( Table 3) provided high quality of monochrome cameras with highresolution and multi-band filter cubes set that provided greater flexibility, to match with the sample.…”

“…Fluorescence scanners support multiple filters using mechanical switching or using tunable LED excitation, similar to confocal microscope, the capture signals are assemble in a compose image 43 . Although, the scanner system can capture the ROI area using one filter at the time or changes the filter at each capture to high channel of co-localization 42 , is still impossible to accelerate the process of the scanning to obtain high quality of images and it is variable depending the methodology used in the scanning that can takes from minutes to serval hours 44 . Analysis software's are also important to extract high quality of information from Table 4) need to accessible, easy with automated capabilities of detection, including tissue segmentation and spatial colocalization cell distribution, critically important to study in particular small samples, such as core needle biopsies or small metastatic tumor samples (Figure 3).…”

Whole slide image cytometry: a novel method to detect abnormal DNA content in Barrett's esophagus

“…A typical tissue specimen can exceed 120 000 × 80 000 pixels in size at × 40 magnification (28 gigabytes of uncompressed data or ∼ 1 GB JPEG compressed 30 ). Whole slide imaging, together with appropriate viewing technology, is beginning to be used for a range of pathology applications, such as pathology education, 31,32 image analysis for biomarker discovery, 33,34 and primary diagnosis. [35][36][37] In tissue-based BE research, some researchers have investigated alternative computerized morphometry and texture measurements as indicators for disease progression.…”