Automated rare sperm identification from low-magnification microscopy images of dissociated microsurgical testicular sperm extraction samples using deep learning

We developed a machine learning algorithm capable of automatically analyzing low-magnification (10) bright-field microscopy images to identify rare sperm in microsurgical testicular sperm extractions with 86% sensitivity.

VOLUME 118, ISSUE 1, P90-99


Ryan Lee, B.A.Sc., Luke Witherspoon, M.D., M.Sc., Meghan Robinson, B.Sc., Jeong Hyun Lee, Ph.D., Simon P. Duffy, Ph.D., Ryan Flannigan, M.D., Hongshen Ma, Ph.D. 



To develop a machine learning algorithm to detect rare human sperm in semen and microsurgical testicular sperm extraction (microTESE) samples using bright-field (BF) microscopy for nonobstructive azoospermia patients.


Spermatozoa were collected from fertile men. Testis biopsies were collected from microTESE samples determined to be clinically negative for sperm. A convolutional neural network based on the U-Net architecture was trained using 35,761 BF image patches with fluorescent ground truth image pairs to segment sperm. The algorithm was validated using 7,663 image patches. The algorithm was tested using 7,663 image patches containing abundant sperm, as well as 7,985 image patches containing rare sperm.


In vitro fertilization center and university laboratories.


Normospermic and nonobstructive azoospermia patients.



Main Outcome Measure(s)

Precision (positive predictive value [PPV]), recall (sensitivity), and F1-score of detected sperm locations.


For sperm-only samples, our algorithm achieved 91% PPV, 95.8% sensitivity, and 93.3% F1-score at ×10 magnification. For dissociated microTESE samples doped with an abundant quantity of sperm, our algorithm achieved 84.0% PPV, 72.7% sensitivity, and 77.9% F1-score. For dissociated microTESE samples doped with rare sperm, our algorithm achieved 84.4% PPV, 86.1% sensitivity, and 85.2% F1-score.


Rare sperm can be detected in patients’ testis biopsy samples for potential subsequent use in in vitro fertilization–intracytoplasmic sperm injection. A machine learning algorithm can use BF images at ×10 magnification to accurately detect sperm locations using automated imaging.