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Abstract 1149: Systems pathology by Optra 30 color multiplex immunofluorescence automated whole slide imaging system integrated with image analysis

Publication ,  Conference
Gholap, A; Jadhav, AP; Somwanshi, S; Kamble, G; Doshi, I; Huang, J
Published in: Cancer Research
July 1, 2019

Background A paradigm shift from single marker immunohistochemistry (IHC) to multiplexing in molecular histopathology has helped better understand the cellular composition and heterogeneity of sub population of cells in tissues. Presently, one needs to manually exchange filters after each multiplex scan for capturing all fluorophores per staining round. This is very cumbersome and inefficient, as this process requires rescanning the same slide multiple times. Moreover, visual assessment and quantification of biomarkers is limited by inter and intra-observer variability. We hereby present an automated multispectral whole-slide imaging system integrated with image analysis for detection of upto 30-color multiplex assay with spectral unmixing and cell segmentation functionality.Material and MethodsMultiplex IHC whole slide imaging Formalin Fixed Paraffin Embedded rat brain; indirect IHC stained sections with 10 antibodies using photostable fluorophores were obtained. The staining was based on antigenic integrity and antibody binding affinity. Optra's multispectral fluorescent whole slide scanning process was optimized to image sensitive antigens in early cycles and robust antigens in latter cycles. The fully automated fluorescence scanning system imaged weakly expressing biomarkers in the visible range (480-620 nm), highly expressing targets in the violet/blue (405-440 nm), far-red (650-680 nm) and NIR (700-800 nm) range across the entire usable light spectrum (350-800 nm).ResultsWe could simultaneously detect up to 30 biomarkers within a single tissue section, using 12 staining-imaging-stripping cycles, with 10 antibodies screened per cycle. Software measures enabled separation of spectrally overlapping markers in the multiplex assay. A computer-controlled X-Y-Z stage used autofocusing at 20X/40X magnification. Integrated image analysis solution performed comprehensive evaluation of antigens of interest with accurate quantification of the target proteins based on parameters including but not limited to hue value/width, intensity threshold scoring, morphological characterization, pixel-count thresholding and color saturation.Conclusion This proposed solution is efficient compared to the current imaging methods to tackle the aforementioned image acquisition and image analysis challenges associated with multiplexing assays.Note: This abstract was not presented at the meeting.Citation Format: Abhi Gholap, Anagha P. Jadhav, Suraj Somwanshi, Gurunath Kamble, Isha Doshi, Jiaoti Huang. Systems pathology by Optra 30 color multiplex immunofluorescence automated whole slide imaging system integrated with image analysis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1149.

Duke Scholars

Published In

Cancer Research

DOI

EISSN

1538-7445

ISSN

0008-5472

Publication Date

July 1, 2019

Volume

79

Issue

13_Supplement

Start / End Page

1149 / 1149

Publisher

American Association for Cancer Research (AACR)

Related Subject Headings

  • Oncology & Carcinogenesis
  • 3211 Oncology and carcinogenesis
  • 3101 Biochemistry and cell biology
  • 1112 Oncology and Carcinogenesis
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Gholap, A., Jadhav, A. P., Somwanshi, S., Kamble, G., Doshi, I., & Huang, J. (2019). Abstract 1149: Systems pathology by Optra 30 color multiplex immunofluorescence automated whole slide imaging system integrated with image analysis. In Cancer Research (Vol. 79, pp. 1149–1149). American Association for Cancer Research (AACR). https://doi.org/10.1158/1538-7445.am2019-1149
Gholap, Abhi, Anagha P. Jadhav, Suraj Somwanshi, Gurunath Kamble, Isha Doshi, and Jiaoti Huang. “Abstract 1149: Systems pathology by Optra 30 color multiplex immunofluorescence automated whole slide imaging system integrated with image analysis.” In Cancer Research, 79:1149–1149. American Association for Cancer Research (AACR), 2019. https://doi.org/10.1158/1538-7445.am2019-1149.
Gholap A, Jadhav AP, Somwanshi S, Kamble G, Doshi I, Huang J. Abstract 1149: Systems pathology by Optra 30 color multiplex immunofluorescence automated whole slide imaging system integrated with image analysis. In: Cancer Research. American Association for Cancer Research (AACR); 2019. p. 1149–1149.
Gholap, Abhi, et al. “Abstract 1149: Systems pathology by Optra 30 color multiplex immunofluorescence automated whole slide imaging system integrated with image analysis.” Cancer Research, vol. 79, no. 13_Supplement, American Association for Cancer Research (AACR), 2019, pp. 1149–1149. Crossref, doi:10.1158/1538-7445.am2019-1149.
Gholap A, Jadhav AP, Somwanshi S, Kamble G, Doshi I, Huang J. Abstract 1149: Systems pathology by Optra 30 color multiplex immunofluorescence automated whole slide imaging system integrated with image analysis. Cancer Research. American Association for Cancer Research (AACR); 2019. p. 1149–1149.

Published In

Cancer Research

DOI

EISSN

1538-7445

ISSN

0008-5472

Publication Date

July 1, 2019

Volume

79

Issue

13_Supplement

Start / End Page

1149 / 1149

Publisher

American Association for Cancer Research (AACR)

Related Subject Headings

  • Oncology & Carcinogenesis
  • 3211 Oncology and carcinogenesis
  • 3101 Biochemistry and cell biology
  • 1112 Oncology and Carcinogenesis