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Improved detection of fluorescently labeled microspheres and vessel architecture with an imaging cryomicrotome.

Due to spectral overlap, the number of fluorescent labels for imaging cryomicrotome detection was limited to 4. The aim of this study was to increase the separation of fluorescent labels. In the new imaging cryomicrotome, the sample is cut in slices of 40 microm. Six images are taken for each cuttin...

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Published in:Medical & Biological Engineering & Computing Vol. 48; no. 8; pp. 735 - 745
Main Authors: van Horssen P, Siebes M, Hoefer I, Spaan JA, van den Wijngaard JP, van Horssen, Pepijn, Siebes, Maria, Hoefer, Imo, Spaan, Jos A E, van den Wijngaard, Jeroen P H M
Format: research Journal Article
Published: Springer Nature Aug2010
Online Access:View this record in EBSCOhost
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      jtl: Medical & Biological Engineering & Computing
      issn: 01400118
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      dt: Aug2010
      vid: 48
      iid: 8
      pid: 237
      pub: Springer Nature
      place: New York, New York
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        104912431
        NLM20574721
        2010713746
        10.1007/s11517-010-0652-8
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      tig:
        atl: Improved detection of fluorescently labeled microspheres and vessel architecture with an imaging cryomicrotome.
      aug:
        au:
          van Horssen P
          Siebes M
          Hoefer I
          Spaan JA
          van den Wijngaard JP
          van Horssen, Pepijn
          Siebes, Maria
          Hoefer, Imo
          Spaan, Jos A E
          van den Wijngaard, Jeroen P H M
        affil: Department of Biomedical Engineering and Physics, Academic Medical Center, University of Amsterdam, Meibergdreef 9, 1105 AZ, Amsterdam, The Netherlands
      sug:
        subj:
          Coronary Circulation Physiology
          Cryoultramicrotomy Methods
          Latex
          Animal Studies
          Dogs
          Fluorescent Dyes Diagnostic Use
          Image Processing, Computer Assisted Methods
          Light
      ab: Due to spectral overlap, the number of fluorescent labels for imaging cryomicrotome detection was limited to 4. The aim of this study was to increase the separation of fluorescent labels. In the new imaging cryomicrotome, the sample is cut in slices of 40 microm. Six images are taken for each cutting plane. Correction for spectral overlap is based on linear combinations of fluorescent images. Locations of microspheres are determined by using the system point spread function. Five differently colored microspheres were injected in vivo distributed over two major coronaries, the left anterior descending and left circumflex artery. Under absence of collateral flow, microspheres outside of target perfusion territories were not found and the procedure did not generate false positive detection when spectral overlap was relevant. In silico-generated microspheres were used to test the effect of background image, transparency correction, and color separation. The percentage of microspheres undetected was 2.3 +/- 0.8% in the presence and 1.5 +/- 0.4% in the absence of background structures with a density of 900 microspheres per color per cm(3). The image analysis method presented here, allows for an increased number of experimental conditions that can be investigated in studies of regional myocardial perfusion.
      pubtype: Academic Journal
      doctype:
        research
        Journal Article
      ougenre: Article
    language: English
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