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Purpose Cornea transplant is among the most successful forms of transplantation; however, contamination of donor corneas poses a significant threat to patient safety, and may result in post-transplant infections and graft failure. To prevent corneal infections and ensure the quality of tissues preserved at 28°C-37°C, specific guidelines have been established to maintain microbiological safety during corneal handling. The methods we employ to prevent the distribution of contaminated corneas include BACTEC™ FX (Becton Dickinson) HB&L® (Alifax) and Gram staining; these approaches guarantee microorganisms detection across various conditions. However, slower-growing microbes such as fungi might evade certain microbiological controls, resulting in the transplantation of contaminated corneas. Consequently, fungi represent a primary concern in eye banking, requiring approaches to detect yeasts and molds particularly when the microbial load is low. Therefore, we are validating a β-D-glucan test (Limusave MT-7500 Fujifilm) to detect fungal contamination in culture media, even when other methods fail.Methods The need to adopt diverse strategies for microbiological safety arises from the varying sensitivity and time demands of each microbiological technique, as well as scheduling constraints imposed by surgical procedures. The HB&L instrument provides rapid results by detecting microorganisms, particularly bacteria, through sample turbidity analysis. While traditional microbiological tests may take several days to yield results, the HB&L system delivers outcomes in just a few hours. However, due to the brief incubation time specified in our protocol, some slow-growing organisms may go undetected. Therefore, it is crucial to utilize systems that support longer incubation periods. The BACTEC™ FX operates by measuring the levels of CO2 produced by microorganisms in specific vials containing broth medium inoculated with cornea medium. We adopted this instrument to monitor microbial growth over extended periods. As a third strategy, we employ Gram staining on the tissue storage medium. This method is particularly valuable to rapidly recognize and partially identify microbes that may evade other control techniques, including fungi, before the conclusion of the culturing process. Unfortunately, the sensitivity of this method represents a significant limitation in cases of low microbial loads. As a novel approach to improve protocols for detecting molds and yeasts, we are currently validating the Limusave MT-7500 instrument that detects fungi by analyzing the levels of (1→3)-β-D-glucan in the sample. This molecule is the most abundant cell wall polysaccharide in fungi and can be detected through a reaction with Limulus amebocyte lysate.Results In most instances, corneal contamination can be effectively detected through the analysis of the storage medium using BACTEC™ FX, HB&L®, and Gram staining. Additionally, the β-D-glucan test for fungal detection was performed on approximately 140 specimens of storage medium, including both positive and negative samples. Currently, the analyses demonstrate that the instrument can reliably detect the presence of fungi in storage media.Conclusions While traditional methods such as BACTEC™ FX, HB&L®, and Gram staining are generally effective against many microorganisms, detecting fungi remains a significant challenge. The β-D-glucan test we are currently validating for fungal contamination appears promising, however further investigation is required to implement this approach in routine microbiological monitoring.