Over the past fifty years, neonatal screening has become essential in the public health programs of a large number of countries. During all these years, the number of detectable diseases has continued to grow, following the possibilities offered by technical advances in clinical biology. The Guthrie test has enabled the miniaturization of blood sampling, opening up the possibilities of biological screening in the newborn population. Fluorimetry, immunoassay and more recently tandem mass spectrometry have subsequently allowed to detect many treatable disorders. The new developments of next generation sequencing and artificial intelligence may open a new era despite many ethical questions that will arise. This review provides an overview of the biological techniques currently used for neonatal screening and opens up perspectives on the place of new technological developments.