Determinants of Test Variability in Scotopic Microperimetry: Effects of Dark Adaptation and Test Indices

Giovanni Montesano, Timos K. Naska, Bethany E. Higgins, David M. Wright, Ruth E. Hogg, David P. Crabb*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

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To test the effect of different dark adaptation conditions and reliability indices on the variability of two color scotopic microperimetry. 


 We analyzed data from 22 consecutive visually healthy adults. Scotopic microperimetry was performed (Macular Integrity Assessment microperimeter, Center-Vue, Padua, Italy) with two wavelength stimuli, cyan (505 nm) and red (627 nm), after a dark adaptation time of 10, 20, or 30 minutes. All tests were repeated twice to measure test–retest variability with Bland–Altman plots. We also provide a method to more accurately quantify the false-positive (FP) responses based on response data (button pressing) from the device, similar to FP responses used in standard static perimetry. Data on fixation stability (95% bivariate contour ellipse area) and blind spot responses were also extracted. Their relationship with measured sensitivity (in decibels) and test–retest variability was quantified through linear mixed effect models.


 Dark adaptation had a significant effect on the sensitivity (dB) measured with the cyan stimulus (P < 0.001), but no effect on the red stimulus. Of the three metrics, the novel FP responses showed the best association with test–retest variability and was the only predictor consistently significant for all tests (P < 0.01).


Dark adaptation protocols should be carefully standardized for scotopic testing, especially if a cyan stimulus is used. The proposed FP responses should be used to assess reliability of microperimetry examinations instead of other metrics. Translational Relevance: We developed a method to calculate a more accurate estimate of the FP responses using data available to all researchers, generalizable to all Macular Integrity Assessment microperimeter tests.

Original languageEnglish
Article number26
Number of pages11
JournalTranslational Vision Science and Technology
Issue number1
Publication statusPublished - 15 Jan 2021

Bibliographical note

Funding Information:
The authors are grateful to all the participants of the NICOLA Study, and the whole NICOLA team, which includes nursing staff, research scientists, research optometrists, clerical staff, computer and laboratory technicians, managers and receptionists. The Atlantic Philanthropies, the Economic and Social Research Council, the UKCRC Centre of Excellence for Public Health Northern Ireland, the Centre for Ageing Research and Development in Ireland, the Office of the First Minister and Deputy First Minister, the Health and Social Care Research and Development Division of the Public Health Agency, the Wellcome Trust/Wolfson Foundation and Queen’s University Belfast provide core financial support for NICOLA. The authors alone are responsible for the interpretation of the data and any views or opinions presented are solely those of the authors and do not necessarily represent those of the NICOLA Study team.

Funding Information:
The NISA study was funded by the College of Optometrists, Macular Society, RNIB, Diabetes UK and the Thomas Pocklington Trust.

Publisher Copyright:
© 2021 The Authors.

Copyright 2021 Elsevier B.V., All rights reserved.


  • Dark adaptation
  • Microperimetry
  • Scotopic sensitivity
  • Test–retest variability

ASJC Scopus subject areas

  • Biomedical Engineering
  • Ophthalmology


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