title: Optimal z-axis scanning parameters for gynecologic cytology specimens.
abstract: BACKGROUND
  The use of virtual microscopy (VM) in clinical cytology has been limited due to the inability to focus through three dimensional (3D) cell clusters with a single focal plane (2D images). Limited information exists regarding the optimal scanning parameters for 3D scanning.
  
  
  OBJECTIVE
  The purpose of this study was to determine the optimal number of the focal plane levels and the optimal scanning interval to digitize gynecological (GYN) specimens prepared on SurePath™ glass slides while maintaining a manageable file size.
  
  
  METHODS
  The iScanCoreo Au scanner (Ventana, AZ, USA) was used to digitize 192 SurePath™ glass slides at three focal plane levels at 1 μ interval. The digitized virtual images (VI) were annotated using BioImagene's Image Viewer. Five participants interpreted the VI and recorded the focal plane level at which they felt confident and later interpreted the corresponding glass slide specimens using light microscopy (LM). The participants completed a survey about their experiences. Inter-rater agreement and concordance between the VI and the glass slide specimens were evaluated.
  
  
  RESULTS
  This study determined an overall high intra-rater diagnostic concordance between glass and VI (89-97%), however, the inter-rater agreement for all cases was higher for LM (94%) compared with VM (82%). Survey results indicate participants found low grade dysplasia and koilocytes easy to diagnose using three focal plane levels, the image enhancement tool was useful and focusing through the cells helped with interpretation; however, the participants found VI with hyperchromatic crowded groups challenging to interpret. Participants reported they prefer using LM over VM. This study supports using three focal plane levels and 1 μ interval to expand the use of VM in GYN cytology.
  
  
  CONCLUSIONS
  Future improvements in technology and appropriate training should make this format a more preferable and practical option in clinical cytology.
authors: Cytotechnology Education, School of Allied Health Professions, University of Nebraska Medical Center, Omaha, Nebraska, USA.
authors: Amber D
authors: AD
authors: Donnelly
authors: Cytotechnology Education, School of Allied Health Professions, University of Nebraska Medical Center, Omaha, Nebraska, USA.
authors: Maheswari S
authors: MS
authors: Mukherjee
authors: Department of Biostatistics, College of Public Health, University of Nebraska Medical Center, Omaha, NE, USA.
authors: Elizabeth R
authors: ER
authors: Lyden
authors: Department of Pathology and Microbiology, College of Medicine, University of Nebraska Medical Center, Omaha, NE, USA.
authors: Julia A
authors: JA
authors: Bridge
authors: Department of Pathology and Microbiology, College of Medicine, University of Nebraska Medical Center, Omaha, NE, USA.
authors: Subodh M
authors: SM
authors: Lele
authors: Department of Cytology, Alegent-Creighton Health, Omaha, NE, USA.
authors: Najia
authors: N
authors: Wright
authors: Department of Cytology, The Nebraska Medical Center, Omaha, NE, USA.
authors: Mary F
authors: MF
authors: McGaughey
authors: Texas Health Hugley Hospital, Fort Worth South, Burleson, TX, USA.
authors: Alicia M
authors: AM
authors: Culberson
authors: Department of Pathology and Microbiology, College of Medicine, University of Nebraska Medical Center, Omaha, NE, USA.
authors: Adam J
authors: AJ
authors: Horn
authors: Department of Pathology and Microbiology, College of Medicine, University of Nebraska Medical Center, Omaha, NE, USA.
authors: Whitney R
authors: WR
authors: Wedel
authors: Department of Pathology and Microbiology, College of Medicine, University of Nebraska Medical Center, Omaha, NE, USA.
authors: Stanley J
authors: SJ
authors: Radio
keywords: Cytology; digital pathology; virtual microscopy; z-axis