Highly sensitive and quick in ovo sexing of domestic chicken eggs by two-wavelength fluorescence spectroscopy

Research output: Contribution to journalResearch articleContributedpeer-review


The in ovo sexing of chicken eggs is a current task and a prerequisite to overcome the mass killing of male day-old chicks from laying lines. Although various methods have been developed and tested in recent years, practicable methods for sex determination are still missing which can be applicated in poultry hatcheries before the chicken embryo is capable of nociception and pain sensation. Optical spectroscopic methods enable an early determination of the sex. In this study, a novel method based on two-wavelength in ovo fluorescence excitation is described. More than 1600 eggs were examined. In ovo fluorescence was sequentially excited at 532 nm and 785 nm. The fluorescence intensities of the spectral regions behave inversely with respect to sex. It is shown that the observed sex-related differences in the fluorescence intensities are based on the embryonic hemoglobin synthesis. The accuracy of sex determination is 96% for both sexes. The hatching rate is not reduced compared to an equivalent reference group. Graphical Abstract: [Figure not available: see fulltext.]


Original languageEnglish
Pages (from-to)603–613
Number of pages11
JournalAnalytical and Bioanalytical Chemistry
Issue number4
Publication statusPublished - 3 Dec 2022

External IDs

Scopus 85143203042
PubMed 36462048
WOS 000893399500001
Mendeley 2bf095fb-2304-3a0c-988a-ebb4f89163cf



  • Fluorescence spectroscopy, Heme synthesis, In ovo sexing, Protoporphyrin IX, Two-wavelength spectroscopy, Male, Chick Embryo, Eggs, Sex Determination Analysis/methods, Animals, Chickens, Female, Ovum, Spectrometry, Fluorescence/methods