Clinical ScienceQuantitative Analysis of Endothelial Cell Loss in Preloaded Descemet Membrane Endothelial Keratoplasty GraftsWolle, Meraf A. MD, MPH*,†; DeMill, David L. MD*,‡; Johnson, Lauren MS§; Lentz, Stephen I. PhD¶; Woodward, Maria A. MD, MS*; Mian, Shahzad I. MD*Author Information *Kellogg Eye Center, University of Michigan, Ann Arbor, MI; †Wilmer Eye Institute, Johns Hopkins University, Baltimore, MD; ‡Devers Eye Institute, Portland, OR; §Eversight Michigan, Ann Arbor, MI; and ¶Department of Internal Medicine, Division of Metabolism, Endocrinology and Diabetes, University of Michigan, MI, USA. Reprints: Shahzad I. Mian, MD, Department of Ophthalmology, University of Michigan, W.K. Kellogg Eye Center, 1000 Wall St, Ann Arbor, MI 48105 (e-mail: [email protected]). Eversight Michigan (M.A.W.), M. A. Woodward receives a grant (K23 Mentored Clinical Scientist Award K23EY023596) from National Institutes of Health. The authors have no conflicts of interest to disclose. Cornea: November 2017 - Volume 36 - Issue 11 - p 1295-1301 doi: 10.1097/ICO.0000000000001301 Buy Metrics Abstract Purpose: Availability of preloaded Descemet membrane endothelial keratoplasty (pDMEK) tissue may increase acceptance of DMEK in surgical management of endothelial disease. The goal of this study was to determine the safety of pDMEK grafts for 24 hours before surgery by analyzing endothelial cell loss (ECL) using 2 image analysis software programs. Methods: A total of 18 cadaveric corneas were prepared for DMEK using a standardized technique and loaded in a modified Jones tube injector. Nine of the corneas were injected into Calcein AM vital dye after 1 minute (controls), and the remaining 9 corneas were left preloaded for 24 hours before injection into vital dye for staining. The stained corneas were imaged using an inverted confocal microscope. ECL was then analyzed and quantified by 2 different graders using 2 image analysis software programs. Results: The control DMEK tissue resulted in 22.0% ± 4.0% ECL compared with pDMEK tissue, which resulted in 19.2% ± 7.2% ECL (P = 0.31). Interobserver agreement was 0.93 for MetaMorph and 0.92 for Fiji. The average time required to process images with MetaMorph was 2 ± 1 minutes and with Fiji was 20 ± 10 minutes. Intraobserver agreement was 0.97 for MetaMorph and 0.93 for Fiji. Conclusions: Preloading DMEK tissue is safe and may provide an alternative technique for tissue distribution and surgery for DMEK. The use of MetaMorph software for quantifying ECL is a novel and accurate imaging method with increased efficiency and reproducibility compared with the previously validated Fiji. Copyright © 2017 Wolters Kluwer Health, Inc. All rights reserved.