ABSTRACT We use images collected with the near-infrared camera (NIRCam) onboard the JWST and with the Hubble Space Telescope (HST) to investigate multiple populations at the bottom of the main sequence (MS) of 47 Tucanae. The mF115W versus mF115W − mF322W2 colour–magnitude diagram (CMD) from NIRCam shows that, below the knee, the MS stars span a wide colour range, where the majority of M-dwarfs exhibit blue colours, and a tail of stars are distributed towards the red. A similar pattern is observed from the mF160W versus mF110W − mF160W CMD from HST, and multiple populations of M-dwarfs are also visible in the optical mF606W versus mF606W − mF814W CMD. The NIRCam CMD shows a narrow sequence of faint MS stars with masses smaller than $0.1\, \mathcal {M}_{\odot }$. We introduce a chromosome map of M-dwarfs that reveals an extended first population and three main groups of second-population stars. By combining isochrones and synthetic spectra with appropriate chemical composition, we simulate colours and magnitudes of different stellar populations in the NIRCam filters (at metallicities [Fe/H] = −1.5 and [Fe/H] = −0.75) and identify the photometric bands that provide the most efficient diagrams to investigate the multiple populations in globular clusters. Models are compared with the observed CMDs of 47 Tucanae to constrain M-dwarfs’ chemical composition. Our analysis suggests that the oxygen range needed to reproduce the colours of first- and second-population M-dwarfs is similar to that inferred from spectroscopy of red giants, constraining the proposal that the chemical variations are due to mass transfer phenomena in proto-clusters.