More specifically, a significant number of studies have focused on introducing new cell-phone based systems that provide low-cost alternatives to conventional microscopy techniques for mHealth applications including malaria detection5,40,41,42,43,44. Mobile phone based microscopy approaches can be broken down into three specific design areas including: lensless approaches40,42, on-lens approaches41 and attachment based approaches5,43,44. Research in each of these design categories has produced promising scientific approaches towards cell-phone microscopy designs capable of significantly affecting healthcare standards in developing countries, particularly in the area of single cell resolution for disease diagnosis. Several cell-phone based microscopy designs for malaria diagnostic applications have been reported.
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To show the contrast polarized light microscopy provides from thin smears of Plasmodium chabaudi malaria-infected blood samples, the images in Fig. 6 are presented. Specifically in Fig. 6A,B, brightfield non-polarized and polarized thin Giemsa-stained blood smear samples of malaria-infected RBCs at 40X magnification images were obtained via a digital SLR camera mounted onto a Leica DMLM polarized microscope. As indicated by the presence of birefringent changes in the polarized reference image, Fig. 6B, the sample had positive infected areas with the malaria-parasite. The presence of hemozoin particles in the sample cause the polarized transmitted illumination light to vary in intensity and wavelength due to variation in the light as it transmits through the birefringent hemozoin particles. The result of this change is represented by seven bright white dots that appear in the cross-polarized reference image. It should be noted that it is very challenging to detect these hemozoin particles in the original non-polarized reference imaging system without being a highly trained technician. This confirms previous reports, by Maude et al. and others, that the use of polarized microscopy in observing the presence of malaria-infected RBCs has shown to improve diagnostic capability up to two fold in some instances7,26. Following the acquisition of the two reference images, two additional images, a non-polarized and polarized image, were capture from the same malaria-infected sample and sample region of the blood smear with the MOPID and are shown in Fig. 6C,D. It is clear from the non-polarized images in Figs 5 and 6A,C that the mobile platform has a reduced system resolution as compared to the reference microscope in polarized mode. However, in examining the polarized images from both systems it is clear that the presence of birefringence appears at the same spots within the sample. This indicates that the results obtained with the MOPID are capable of determining the presence of malaria with lower system resolution and with less user expertise than traditional microscopy requires.
The portable commercial microscope lens assembly consisted of two separate plastic lens modules and had a numerical aperture (NA) value of 0.65. The MOPID design incorporated white light LEDs placed at a distance from the sample, allowing for even field illumination. The individual LEDs were chosen because of their low-cost, low power, durability and long lifetime; all characteristics ideally suited for use in low-resource settings5,13,58,59. In the designed system, the white light LED total power was 66 lumens after the light passes through a TiO2 diffuser plate. The diffuser plate is utilized to allow for homogenous illumination across the sample and was followed by a polarizer sheet generating linearly polarized light prior to transmission through the sample. The opening in Fig. 1B, labeled slide rack, is the location where the blood smear or histological slide is inserted into the optical train. This slit is located such that the sample is optimally positioned at approximately one focal length of the imaging system. Utilizing a multi-position insert the slide can be manually moved past the camera from left to right in incremental steps. The final component prior to the mobile phone surface in the optical path is a second polarizer, an analyzer, capable of being oriented either at 45 or 90 degrees with respect to the initial polarizer orientation.
Metafer managed imaging systems scan specimen of various sizes, use many different contrasting methods and magnifications, and find many different target objects. Due to Metafer's modularity and its flexible architecture, it is applicable to numerous applications, including cytogenetic diagnostics, hematology, pathology, toxicology, forensic sciences, and many more.
Legal NoteMetaSystems software and system products are classified as in vitro diagnostic medical devices (IVD) in the European Union in accordance with In Vitro Diagnostics Regulation (EU) 2017/746 or In Vitro Diagnostic Medical Device Directive 98/79/EC, respectively, and carry the CE label unless otherwise indicated. Use all MetaSystems IVD products only within the scope of their intended purpose.
The system uses precisely tailored pulses of light to simultaneously image with multiple wavelengths. This enables the researchers to study concurrent processes within cells and tissue, and could give cancer researchers a new tool for tracking tumor progression and physicians new technology for tissue pathology and diagnostics.
Clinical decision-making in breast cancer increasingly depends on the quantitative evaluation of molecular and immunohistochemical markers, such as Ki67, for tumor proliferation assessment. However, recent studies have shown a limited reliability of the established manual Ki67 scoring in breast cancer diagnostics in a multicenter setting. As a solution, we present a novel automated Ki67 scoring system, which we validate using data from the prospective neoadjuvant GeparTrio trial. Our approach is readily deployable in routine diagnostics and may thus help solve the recently reported reliability issues in Ki67 scoring.
Core-needle biopsies of breast tumors from patients acquired in a routine diagnostic setting that were formalin-fixed and paraffin-embedded were used in this study. Tissue specimens were fixated at room temperature in 10% neutral-buffered formalin immediately after biopsies were acquired (maximum time to fixation 3 minutes). Time of fixation was 16 to 72 hours. Subsequently, specimens were paraffin-embedded and further processed on the same or the following day.
Interestingly, the overall results are relatively similar between manual and automated approaches, which might appear surprising given the high variability of manual Ki67 scoring reported previously (10). However, this is likely due to the fact that the manual scoring results were obtained by using a standardized stringent monocentric evaluation with exact counting of cells rather than semiquantitative estimations. Therefore, automated Ki67 scoring can be expected to perform more robustly than manual approaches in routine diagnostics where a considerable intra- and interobserver variability has been observed for manual scoring (9). But even if manual scoring performed similarly also in routine settings, would our automated approach have several advantages over manual scoring: Whereas exact manual counting of about 500 cells takes between about 6 and 10 minutes, our approach requires about 30 to 40 seconds to score Ki67 in a typical routine diagnostic field of view. Similarly, the automated system is capable of evaluating the complete cohort of over 1,000 cases within a day, which is not feasible manually.
Regarding the broader use of our approach in routine diagnostics, recent implementation at other sites also appears to be confirming the robustness of our method, and another systematic study addressing automated Ki67 scoring performance under routine diagnostic conditions, including the analysis of samples processed in different laboratories, is currently under way.
To summarize, our approach allows for a robust and standardized fully automated Ki67 scoring that may therefore contribute to a solution to the widely criticized variations observed in current manual evaluation that have fundamentally put the utility of Ki67 as a prognostic and predictive biomarker in breast cancer diagnostics into question.
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Patients with suspected meningitis are treated empirically pending diagnostic results.3,4 This can mean lengthy hospitalizations and unnecessary antimicrobial use, all of which add to the overall cost of care.
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