Fluorescence imaging for whole slide scanning using LED-based color sequential illumination

Leon Van Der Graaff, Fanny Boyaval, Lucas J. Van Vliet, Sjoerd Stallinga

Research output: Chapter in Book/Conference proceedings/Edited volumeConference contributionScientificpeer-review

7 Citations (Scopus)
64 Downloads (Pure)


In the field of pathology there is an ongoing transition to the use of Whole Slide Imaging (WSI) systems which scan tissue slides at intermediate resolution (0∼.25 μm) and high throughput (15mm2=min) to digital image files. Most scanners currently on the market are line-sensor based push-broom scanners for three-color (RGB) brightfield imaging. Adding the ability of fluorescence imaging opens up a wide range of possibilities to the field, in particular the use of specific molecular (proteins, genes) imaging techniques. We propose an extension to fluorescence imaging for a highly efficient WSI systems based on a line scanning technique using multi-color led epi-illumination. The use of multi-band dichroics eliminates the need for filter wheels or any other moving parts in the system, the use of color sequential illumination with leds enables imaging of multiple color channels with a single sensor. Our approach offers a solution to fluorescence WSI systems that is technologically robust and cost-effective. We present design details of a four-color led based epi-illumination with a quad-band dichroic filter optimized for leds. We provide a thorough analysis regarding the obtained optical and spectral efficiency. The primary throughput limitation is the minimum Signal-to-Noise-Ratio (SNR) given the available optical power in the illumination etendue, and indicates that a throughput on the order of 1000 lines/sec can be obtained.

Original languageEnglish
Title of host publicationOptics, Photonics, and Digital Technologies for Imaging Applications V
EditorsP. Schelkens, T. Ibrahimi, G. Cristobal
PublisherSociety of Photo-Optical Instrumentation Engineers
Number of pages15
ISBN (Print)978-151061884-8
Publication statusPublished - 2018
EventOptics, Photonics, and Digital Technologies for Imaging Applications V 2018 - Strasbourg, France
Duration: 24 Apr 201826 Apr 2018


ConferenceOptics, Photonics, and Digital Technologies for Imaging Applications V 2018

Bibliographical note

Green Open Access added to TU Delft Institutional Repository ‘You share, we take care!’ – Taverne project https://www.openaccess.nl/en/you-share-we-take-care Otherwise as indicated in the copyright section: the publisher is the copyright holder of this work and the author uses the Dutch legislation to make this work public.


  • Digital Pathology
  • Epi-illumination
  • Fluorescence Imaging
  • LED
  • Microscopy
  • Scanning
  • Whole Slide Imaging (WSI)


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