Abstract
Obtaining consistent photomicrographic images of pathology slides is not always easy because of many different types and settings of the equipment such as the microscopes and digital cameras. In this study, we developed a photomicrography technique that could acquire consistent images of pathology slides. The neutral density (ND) filter was attached to a transparent glass slide as a reference slide, photographed using consistent settings, and acquired images that harbored all of the areas of gray, white, and black. In the same way, the slide was replaced by the actual pathology slide, and photomicrographed. To simulate different light environment, the above photographic session was repeated using two different light intensities and microscopes. A graphic program was used to adjust levels of the reference slide images and this leveling, or calibration, was saved as a file for each. This file for leveling process was applied to actual subsequent photomicrographic images. The same sites of noncalibrated and calibrated images of the pathology slide were calculated into CIELAB or CIE L*a* coordinates. Then, the color differences (ΔE*ab) were calculated. As results, in the study using a 50% transmittance ND filter, two original different images were made nearly identical to the unaided eye, especially in two-point (white and gray) and three-point (black, white, and gray) leveling. In comparison of different light intensities, the ΔE*ab of the selected area was 0.9 in two-point leveling. Between different microscopes, 10.7 of ΔE*ab was the smallest value in three-point leveling. This method would be helpful for acquiring consistent photomicrographic images of pathology slides.
| Original language | English |
|---|---|
| Pages (from-to) | 397-400 |
| Number of pages | 4 |
| Journal | Microscopy Research and Technique |
| Volume | 74 |
| Issue number | 5 |
| DOIs | |
| Publication status | Published - 2011 May 1 |
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Keywords
- Image processing
- Microscopy
- Pathology
- Photomicrography
ASJC Scopus subject areas
- Anatomy
- Instrumentation
- Histology
- Medical Laboratory Technology
Cite this
A novel consistent photomicrography technique using a reference slide made of neutral density filter. / Kim, Jong Yeob; Kim, Ji Woong; Seo, Soo-Hong; Kye, Young Chul; Ahn, Hyo Hyun.
In: Microscopy Research and Technique, Vol. 74, No. 5, 01.05.2011, p. 397-400.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - A novel consistent photomicrography technique using a reference slide made of neutral density filter
AU - Kim, Jong Yeob
AU - Kim, Ji Woong
AU - Seo, Soo-Hong
AU - Kye, Young Chul
AU - Ahn, Hyo Hyun
PY - 2011/5/1
Y1 - 2011/5/1
N2 - Obtaining consistent photomicrographic images of pathology slides is not always easy because of many different types and settings of the equipment such as the microscopes and digital cameras. In this study, we developed a photomicrography technique that could acquire consistent images of pathology slides. The neutral density (ND) filter was attached to a transparent glass slide as a reference slide, photographed using consistent settings, and acquired images that harbored all of the areas of gray, white, and black. In the same way, the slide was replaced by the actual pathology slide, and photomicrographed. To simulate different light environment, the above photographic session was repeated using two different light intensities and microscopes. A graphic program was used to adjust levels of the reference slide images and this leveling, or calibration, was saved as a file for each. This file for leveling process was applied to actual subsequent photomicrographic images. The same sites of noncalibrated and calibrated images of the pathology slide were calculated into CIELAB or CIE L*a* coordinates. Then, the color differences (ΔE*ab) were calculated. As results, in the study using a 50% transmittance ND filter, two original different images were made nearly identical to the unaided eye, especially in two-point (white and gray) and three-point (black, white, and gray) leveling. In comparison of different light intensities, the ΔE*ab of the selected area was 0.9 in two-point leveling. Between different microscopes, 10.7 of ΔE*ab was the smallest value in three-point leveling. This method would be helpful for acquiring consistent photomicrographic images of pathology slides.
AB - Obtaining consistent photomicrographic images of pathology slides is not always easy because of many different types and settings of the equipment such as the microscopes and digital cameras. In this study, we developed a photomicrography technique that could acquire consistent images of pathology slides. The neutral density (ND) filter was attached to a transparent glass slide as a reference slide, photographed using consistent settings, and acquired images that harbored all of the areas of gray, white, and black. In the same way, the slide was replaced by the actual pathology slide, and photomicrographed. To simulate different light environment, the above photographic session was repeated using two different light intensities and microscopes. A graphic program was used to adjust levels of the reference slide images and this leveling, or calibration, was saved as a file for each. This file for leveling process was applied to actual subsequent photomicrographic images. The same sites of noncalibrated and calibrated images of the pathology slide were calculated into CIELAB or CIE L*a* coordinates. Then, the color differences (ΔE*ab) were calculated. As results, in the study using a 50% transmittance ND filter, two original different images were made nearly identical to the unaided eye, especially in two-point (white and gray) and three-point (black, white, and gray) leveling. In comparison of different light intensities, the ΔE*ab of the selected area was 0.9 in two-point leveling. Between different microscopes, 10.7 of ΔE*ab was the smallest value in three-point leveling. This method would be helpful for acquiring consistent photomicrographic images of pathology slides.
KW - Image processing
KW - Microscopy
KW - Pathology
KW - Photomicrography
UR - http://www.scopus.com/inward/record.url?scp=79954558934&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=79954558934&partnerID=8YFLogxK
U2 - 10.1002/jemt.20921
DO - 10.1002/jemt.20921
M3 - Article
C2 - 20812249
AN - SCOPUS:79954558934
VL - 74
SP - 397
EP - 400
JO - Microscopy Research and Technique
JF - Microscopy Research and Technique
SN - 1059-910X
IS - 5
ER -