Aim The aim was to evaluate the efficacy of a combination of intrastromal injections of amphotericin B (AmB) and autologous serum eye drops (ASED) in the treatment of resistant fungal keratitis. Patients and methods This is a prospective nonrandomized study that evaluated 15 eyes of 15 patients with resistant fungal corneal ulcers not responding to conventional medical treatment treated with intrastromal AmB injection (25 µg/ml) combined with the use of ASED. The efficacy, complications, and outcomes were evaluated. Results Patients showed marked improvement in symptoms (e.g. pain, discharge, photophobia, and lacrimation) in 12 cases (80%) by the second week after injection and total disappearance of symptoms in all patients by the second month. By the second week, three eyes (20%) showed complete ulcer healing, at the first month eight eyes (53.3%), at the second month 12 eyes (80%), and at the third month 15 eyes (100%) showed complete ulcer healing. The procedure was performed successfully, and no severe intraoperative or postoperative complications were observed except two patients develop minimal intrastromal bleeding which resolved. All the study cases underwent use of ASED after the intrastromal injection to accelerate ulcer healing, alleviate corneal irritation, and reduce inflammatory reactions and all of them showed rapid improvement of symptoms, for example, pain and good ulcer-healing process. Conclusion Combination of intrastromal injection of AmB and autologous serum eye drops is a promising, safe, and effective technique for the management of resistant fungal corneal ulcers not responding to conventional medical treatment. ASED shows good results in improving epithelial healing, alleviating corneal irritation, and in reducing inflammatory reactions.