Real-time, high-definition, three-dimensional microscopy for evaluating problematic cervical Papanicolaou smears classified as atypical squamous cells of undetermined significance

Abstract

BACKGROUND The perceived inadequacies of the cervical Papanicolaou (Pap) smear have been attributed to sampling, screening, or interpretive errors. Within this type of cytologic preparation, there are thick cell clusters in which the cells are obscured. It may not possible to evaluate these areas by conventional microscopy. The authors clinically tested the hypothesis that high‐definition, three‐dimensional (3‐D) microscopy based on multiple oblique illumination (MOI), with its ability to penetrate into thick areas, would be useful in evaluating problematic cervical Pap smears, particularly those diagnosed as atypical squamous cells of undetermined significance (ASCUS). METHODS ASCUS Pap smears and corresponding surgical biopsy specimens were evaluated prospectively using standard, axially illuminated microscopes and a new high‐definition, 3‐D microscope employing MOI. The Pap smears were reviewed in a blinded fashion with both types of microscopy. The rendered diagnoses were then compared with the subsequent tissue biopsies, which also were blinded, as the definitive end point. RESULTS It was immediately apparent that the high‐definition, 3‐D MOI microscope had better resolution compared with the standard microscopes. Pap smears and biopsy diagnoses were correlated significantly for MOI (P < 0.001), and there were significant improvements (P = 0.0108) in accuracy when 3‐D, high‐definition microscopy was compared with conventional microscopy. The authors found no statistically significant correlation between ASCUS diagnoses that were rendered by using standard microscopes compared with the subsequent biopsy. CONCLUSIONS Due to enhanced visualization through thick cell clusters, an increased depth of field, light penetration, and resolution, high‐definition, 3‐D microscopy based on MOI produced superior accuracy compared with conventional light microscopy in evaluating cervical Pap smears. Cancer (Cancer Cytopathol) 2002. © 2002 American Cancer Society.

Quantification of intraepidermal nerve fiber density using three‐dimensional microscopy

“…Moreover, the device could extend the range of the image generated by LM from two dimensions to three dimensions through z‐stacked images and anaglyphic pictures. The advantages of stereo imaging using 3D microscopy have been reported to be prominent when thick biological preparations are investigated in the fields of neurobiology, vascular biology, and cytopathology, among others (Greenberg & Boyde, ; Ramsamooj et al, ).…”

“…Therefore, 3D microscopy has been considered as a reasonable candidate to overcome the numerous limitations of conventional LM originating from thin sections, flat images, and consequent lack of depth information. The drawbacks of ordinary LM can lead to misinterpretation of clinical samples from patients and contribute to increasing the risk of misdiagnosis (Ramsamooj et al, ).…”

“…As a result, 3D microscopy provides direct stereoscopic views of thick microscopic sections from certain tissues with complex organization such as the central nervous system (Greenberg & Boyde, 1993). Furthermore, superior depth of field of the device in thick areas has been reported to be useful in clinical assessment, such as gynecological evaluation of cervical cancer with pap smear (Ramsamooj, Doolin, Greenberg, Catalano, & Hewitt, 2002).…”

Application of Three-Dimensional Light Microscopy for Thick Specimen Studies

“…Using these characteristics of 3D microscopy, Golgi stained block was observed together with samples such as neuron, glia, and blood vessel (Greenberg & Boyde, 1993). A research has introduced a method that increases accuracy of pap smear pathology by applying 3D microscopy (Ramsamooj et al, 2002). In this research, well focused image was obtained from not only neuron tissue but also from hard tissues in body such as bone and teeth, so further application to hard tissue research seems to be possible.…”

A New Look at Lunar Soil Collected from the Sea of Tranquility during the Apollo 11 Mission

“…In other words, both eyes receive a two-dimensional (2D) image but from a slightly different angle; the human brain then naturally creates the third dimension. This approach has been used in a wide variety of applications, ranging from cancer screening (Ramsamooj et al, 2002) to a comparison of the morphology of sand grains from around the world (Greenberg, 2008). Using oblique lighting in conjunction with the collection and subsequent processing of a through-focal series of frames has allowed us to obtain high-resolution color micrographs of lunar regolith particles showing detail never thought possible with an optical microscope.…”