High-speed spectral nanocytology for early cancer screening

Abstract

Abstract. High-throughput partial wave spectroscopy (HTPWS) is introduced as a high-speed spectral nanocytology technique that utilizes the field effect of carcinogenesis to perform minimally invasive cancer screening on atrisk populations. HTPWS uses fully automated hardware and an acousto-optic tunable filter to scan slides at low magnification, to select cells, and to rapidly acquire spectra at each spatial pixel in a cell between 450 and 700 nm, completing measurements of 30 cells in 40 min. Statistical quantitative analysis on the size and density of intracellular nanostructures extracted from the spectra at each pixel in a cell yields the diagnostic biomarker, disorder strength (L d ). Linear correlation between L d and the length scale of nanostructures was measured in phantoms with R 2 ¼ 0.93. Diagnostic sensitivity was demonstrated by measuring significantly higher L d from a human colon cancer cell line (HT29 control vector) than a less aggressive variant (epidermal growth factor receptor knockdown). Clinical diagnostic performance for lung cancer screening was tested on 23 patients, yielding a significant difference in L d between smokers and cancer patients, p ¼ 0.02 and effect size ¼ 1.00. The high-throughput performance, nanoscale sensitivity, and diagnostic sensitivity make HTPWS a potentially clinically relevant modality for risk stratification of the large populations at risk of developing cancer. © The Authors. Published by SPIE under a Creative Commons Attribution 3.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.

Nanoscale refractive index fluctuations detected via sparse spectral microscopy

“…Each of the proposed markers were also tested experimentally using data previously collected on the High-Throughput PWS system for full-spectrum analysis [13]. This included a casecontrol cell line experiment and two human-subjects cancer screening experiments.…”

“…Buccal cells were collected by brushing from each patient's cheek and these brushings were then smeared onto a glass slide before being fixed in 95% ethanol and stained using Papanicolaou stain [13]. The second human subjects experiment included data from a prostate cancer risk assessment experiment.…”

“…To collect high-resolution spectral data cubes (x,y,λ), a recently developed High-Throughput version of the PWS microscope was used [13]. This instrument used an AOTF (HSI-300, Gooch and Housego, Orlando, Florida) for spectral filtration of the Epi-illumination light from a Xenon lamp (66907-100XF-R1, Newport Corporation, Irvine, California) in combination with a high-speed collection camera (Hamamatsu ORCA Flash 2.8, Bridgewater, New Jersey).…”

“…As a result, performance of the technique has been limited, even with the current high-throughput version of the instrument, which utilizes an acousto-optic tunable filter (AOTF) for high speed spectral filtering [13]. To improve the performance of PWS microscopy and extend its capabilities to measurements of larger samples such as tissue sections, a whole-slide imaging approach has been developed.…”

Reconstruction of explicit structural properties at the nanoscale via spectroscopic microscopy

“…For SM experimental measurements, we used our in-house instrument previously built for high-throughput partial wave spectroscopy measurements: 17 epi-illumination bright-field microscope with small illumination NA ¼ 0.15, moderate collection NA ¼ 0.6, and 40× magnification (objective lens from LUCPlanFL N, Olympus, Center Valley, Pennsylvania). Koehler illumination scheme was implemented for uniformity of incident light intensity throughout the image.…”