Multimodal confocal mosaics enable high sensitivity and specificity in screening of in situ squamous cell carcinoma

Maria Del Carmen Grados Luyando, Anna Bar, Nicholas Snavely, Steven Jacques, Daniel S. Gareau

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

Screening cancer in excision margins with confocal microscopy may potentially save time and cost over the gold standard histopathology (H and E). However, diagnostic accuracy requires sufficient contrast and resolution to reveal pathological traits in a growing set of tumor types. Reflectance mode images structural details due to microscopic refractive index variation. Nuclear contrast with acridine orange fluorescence provides enhanced diagnostic value, but fails for in situ squamous cell carcinoma (SCC), where the cytoplasm is important to visualize. Combination of three modes [eosin (Eo) fluorescence, reflectance (R) and acridine orange (AO) fluorescence] enable imaging of cytoplasm, collagen and nuclei respectively. Toward rapid intra-operative pathological margin assessment to guide staged cancer excisions, multimodal confocal mosaics can image wide surgical margins (∼1cm) with sub-cellular resolution and mimic the appearance of conventional H and E. Absorption contrast is achieved by alternating the excitation wavelength: 488nm (AO fluorescence) and 532nm (Eo fluorescence). Superposition and false-coloring of these modes mimics H and E, enabling detection of the carcinoma in situ in the epidermal layer The sum mosaic Eo+R is false-colored pink to mimic eosins' appearance in H and E, while the AO mosaic is false-colored purple to mimic hematoxylins' appearance in H and E. In this study, mosaics of 10 Mohs surgical excisions containing SCC in situ and 5 containing only normal tissue were subdivided for digital presentation equivalent to 4X histology. Of the total 16 SCC in situ multimodal mosaics and 16 normal cases presented, two reviewers made 1 and 2 (respectively) type-2 errors (false positives) but otherwise scored perfectly when using the confocal images to screen for the presence of SCC in situ as compared to the gold standard histopathology. Limitations to precisely mimic H and E included occasional elastin staining by AO. These results suggest that confocal mosaics may effectively guide staged SCC excisions in skin and other tissues.

Original languageEnglish (US)
Title of host publicationProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Volume8937
DOIs
StatePublished - 2014
EventMultimodal Biomedical Imaging IX - San Francisco, CA, United States
Duration: Feb 1 2014Feb 2 2014

Other

OtherMultimodal Biomedical Imaging IX
CountryUnited States
CitySan Francisco, CA
Period2/1/142/2/14

Fingerprint

Acridine Orange
Carcinoma in Situ
Squamous Cell Carcinoma
Eosine Yellowish-(YS)
Screening
screening
cancer
Fluorescence
Sensitivity and Specificity
sensitivity
fluorescence
Cytoplasm
margins
cytoplasm
Tissue
Elastin
Refractometry
Histology
Confocal microscopy
Optical Imaging

ASJC Scopus subject areas

  • Atomic and Molecular Physics, and Optics
  • Electronic, Optical and Magnetic Materials
  • Biomaterials
  • Radiology Nuclear Medicine and imaging

Cite this

Grados Luyando, M. D. C., Bar, A., Snavely, N., Jacques, S., & Gareau, D. S. (2014). Multimodal confocal mosaics enable high sensitivity and specificity in screening of in situ squamous cell carcinoma. In Progress in Biomedical Optics and Imaging - Proceedings of SPIE (Vol. 8937). [893704] https://doi.org/10.1117/12.2040542

Multimodal confocal mosaics enable high sensitivity and specificity in screening of in situ squamous cell carcinoma. / Grados Luyando, Maria Del Carmen; Bar, Anna; Snavely, Nicholas; Jacques, Steven; Gareau, Daniel S.

Progress in Biomedical Optics and Imaging - Proceedings of SPIE. Vol. 8937 2014. 893704.

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Grados Luyando, MDC, Bar, A, Snavely, N, Jacques, S & Gareau, DS 2014, Multimodal confocal mosaics enable high sensitivity and specificity in screening of in situ squamous cell carcinoma. in Progress in Biomedical Optics and Imaging - Proceedings of SPIE. vol. 8937, 893704, Multimodal Biomedical Imaging IX, San Francisco, CA, United States, 2/1/14. https://doi.org/10.1117/12.2040542
Grados Luyando MDC, Bar A, Snavely N, Jacques S, Gareau DS. Multimodal confocal mosaics enable high sensitivity and specificity in screening of in situ squamous cell carcinoma. In Progress in Biomedical Optics and Imaging - Proceedings of SPIE. Vol. 8937. 2014. 893704 https://doi.org/10.1117/12.2040542
Grados Luyando, Maria Del Carmen ; Bar, Anna ; Snavely, Nicholas ; Jacques, Steven ; Gareau, Daniel S. / Multimodal confocal mosaics enable high sensitivity and specificity in screening of in situ squamous cell carcinoma. Progress in Biomedical Optics and Imaging - Proceedings of SPIE. Vol. 8937 2014.
@inproceedings{57d45da73c9c494abb7cca8d7190e72e,
title = "Multimodal confocal mosaics enable high sensitivity and specificity in screening of in situ squamous cell carcinoma",
abstract = "Screening cancer in excision margins with confocal microscopy may potentially save time and cost over the gold standard histopathology (H and E). However, diagnostic accuracy requires sufficient contrast and resolution to reveal pathological traits in a growing set of tumor types. Reflectance mode images structural details due to microscopic refractive index variation. Nuclear contrast with acridine orange fluorescence provides enhanced diagnostic value, but fails for in situ squamous cell carcinoma (SCC), where the cytoplasm is important to visualize. Combination of three modes [eosin (Eo) fluorescence, reflectance (R) and acridine orange (AO) fluorescence] enable imaging of cytoplasm, collagen and nuclei respectively. Toward rapid intra-operative pathological margin assessment to guide staged cancer excisions, multimodal confocal mosaics can image wide surgical margins (∼1cm) with sub-cellular resolution and mimic the appearance of conventional H and E. Absorption contrast is achieved by alternating the excitation wavelength: 488nm (AO fluorescence) and 532nm (Eo fluorescence). Superposition and false-coloring of these modes mimics H and E, enabling detection of the carcinoma in situ in the epidermal layer The sum mosaic Eo+R is false-colored pink to mimic eosins' appearance in H and E, while the AO mosaic is false-colored purple to mimic hematoxylins' appearance in H and E. In this study, mosaics of 10 Mohs surgical excisions containing SCC in situ and 5 containing only normal tissue were subdivided for digital presentation equivalent to 4X histology. Of the total 16 SCC in situ multimodal mosaics and 16 normal cases presented, two reviewers made 1 and 2 (respectively) type-2 errors (false positives) but otherwise scored perfectly when using the confocal images to screen for the presence of SCC in situ as compared to the gold standard histopathology. Limitations to precisely mimic H and E included occasional elastin staining by AO. These results suggest that confocal mosaics may effectively guide staged SCC excisions in skin and other tissues.",
author = "{Grados Luyando}, {Maria Del Carmen} and Anna Bar and Nicholas Snavely and Steven Jacques and Gareau, {Daniel S.}",
year = "2014",
doi = "10.1117/12.2040542",
language = "English (US)",
isbn = "9780819498502",
volume = "8937",
booktitle = "Progress in Biomedical Optics and Imaging - Proceedings of SPIE",

}

TY - GEN

T1 - Multimodal confocal mosaics enable high sensitivity and specificity in screening of in situ squamous cell carcinoma

AU - Grados Luyando, Maria Del Carmen

AU - Bar, Anna

AU - Snavely, Nicholas

AU - Jacques, Steven

AU - Gareau, Daniel S.

PY - 2014

Y1 - 2014

N2 - Screening cancer in excision margins with confocal microscopy may potentially save time and cost over the gold standard histopathology (H and E). However, diagnostic accuracy requires sufficient contrast and resolution to reveal pathological traits in a growing set of tumor types. Reflectance mode images structural details due to microscopic refractive index variation. Nuclear contrast with acridine orange fluorescence provides enhanced diagnostic value, but fails for in situ squamous cell carcinoma (SCC), where the cytoplasm is important to visualize. Combination of three modes [eosin (Eo) fluorescence, reflectance (R) and acridine orange (AO) fluorescence] enable imaging of cytoplasm, collagen and nuclei respectively. Toward rapid intra-operative pathological margin assessment to guide staged cancer excisions, multimodal confocal mosaics can image wide surgical margins (∼1cm) with sub-cellular resolution and mimic the appearance of conventional H and E. Absorption contrast is achieved by alternating the excitation wavelength: 488nm (AO fluorescence) and 532nm (Eo fluorescence). Superposition and false-coloring of these modes mimics H and E, enabling detection of the carcinoma in situ in the epidermal layer The sum mosaic Eo+R is false-colored pink to mimic eosins' appearance in H and E, while the AO mosaic is false-colored purple to mimic hematoxylins' appearance in H and E. In this study, mosaics of 10 Mohs surgical excisions containing SCC in situ and 5 containing only normal tissue were subdivided for digital presentation equivalent to 4X histology. Of the total 16 SCC in situ multimodal mosaics and 16 normal cases presented, two reviewers made 1 and 2 (respectively) type-2 errors (false positives) but otherwise scored perfectly when using the confocal images to screen for the presence of SCC in situ as compared to the gold standard histopathology. Limitations to precisely mimic H and E included occasional elastin staining by AO. These results suggest that confocal mosaics may effectively guide staged SCC excisions in skin and other tissues.

AB - Screening cancer in excision margins with confocal microscopy may potentially save time and cost over the gold standard histopathology (H and E). However, diagnostic accuracy requires sufficient contrast and resolution to reveal pathological traits in a growing set of tumor types. Reflectance mode images structural details due to microscopic refractive index variation. Nuclear contrast with acridine orange fluorescence provides enhanced diagnostic value, but fails for in situ squamous cell carcinoma (SCC), where the cytoplasm is important to visualize. Combination of three modes [eosin (Eo) fluorescence, reflectance (R) and acridine orange (AO) fluorescence] enable imaging of cytoplasm, collagen and nuclei respectively. Toward rapid intra-operative pathological margin assessment to guide staged cancer excisions, multimodal confocal mosaics can image wide surgical margins (∼1cm) with sub-cellular resolution and mimic the appearance of conventional H and E. Absorption contrast is achieved by alternating the excitation wavelength: 488nm (AO fluorescence) and 532nm (Eo fluorescence). Superposition and false-coloring of these modes mimics H and E, enabling detection of the carcinoma in situ in the epidermal layer The sum mosaic Eo+R is false-colored pink to mimic eosins' appearance in H and E, while the AO mosaic is false-colored purple to mimic hematoxylins' appearance in H and E. In this study, mosaics of 10 Mohs surgical excisions containing SCC in situ and 5 containing only normal tissue were subdivided for digital presentation equivalent to 4X histology. Of the total 16 SCC in situ multimodal mosaics and 16 normal cases presented, two reviewers made 1 and 2 (respectively) type-2 errors (false positives) but otherwise scored perfectly when using the confocal images to screen for the presence of SCC in situ as compared to the gold standard histopathology. Limitations to precisely mimic H and E included occasional elastin staining by AO. These results suggest that confocal mosaics may effectively guide staged SCC excisions in skin and other tissues.

UR - http://www.scopus.com/inward/record.url?scp=84896774083&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84896774083&partnerID=8YFLogxK

U2 - 10.1117/12.2040542

DO - 10.1117/12.2040542

M3 - Conference contribution

AN - SCOPUS:84896774083

SN - 9780819498502

VL - 8937

BT - Progress in Biomedical Optics and Imaging - Proceedings of SPIE

ER -