Healthc Inform Res.  2016 Jul;22(3):238-242. 10.4258/hir.2016.22.3.238.

Methods of Hematoxylin and Erosin Image Information Acquisition and Optimization in Confocal Microscopy

• Yoon WB
• Kim H
• Kim KG
• Choi Y
• Chang HJ
• Sohn DK

• KMID: 2357402
• DOI: http://doi.org/10.4258/hir.2016.22.3.238

Abstract

OBJECTIVES
We produced hematoxylin and eosin (H&E) staining-like color images by using confocal laser scanning microscopy (CLSM), which can obtain the same or more information in comparison to conventional tissue staining.
METHODS
We improved images by using several image converting techniques, including morphological methods, color space conversion methods, and segmentation methods.
RESULTS
An image obtained after image processing showed coloring very similar to that in images produced by H&E staining, and it is advantageous to conduct analysis through fluorescent dye imaging and microscopy rather than analysis based on single microscopic imaging.
CONCLUSIONS
The colors used in CLSM are different from those seen in H&E staining, which is the method most widely used for pathologic diagnosis and is familiar to pathologists. Computer technology can facilitate the conversion of images by CLSM to be very similar to H&E staining images. We believe that the technique used in this study has great potential for application in clinical tissue analysis.

Keyword

Confocal Laser Scanning Microscopy; Computer-Assisted Image Processing; Fluorescence; Histological Labeling; Hematoxylin

MeSH Terms

Diagnosis
Eosine Yellowish-(YS)
Fluorescence
Hematoxylin*
Image Processing, Computer-Assisted
Methods*
Microscopy
Microscopy, Confocal*
Staining and Labeling
Eosine Yellowish-(YS)
Hematoxylin

Figure

• 

  Figure 1 (A) Pictures of H&E stained tumor section, (B) fluorescence image of tumor section obtained using confocal laser scanning microscopy.

• 

  Figure 2 Images obtained during image processing procedure: (A) original fluorescence image, (B, C) conversion from RGB to CIE Luv, (D) merged images, (E) background image conversion, (F) nucleus creation image, (G) enhanced image, (H) mask creation image, and (I) final image.

• 

  Figure 3 Magnified image after the image processing procedure. Nuclei are blue, while cytoplasm and extracellular matrix are pink.

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