In this investigation, ferric (Fe) and nickel (Ni) co-doped tin oxide (SnO2) nanoparticles structural, optical, morphological, and antibacterial characteristics were synthesised, characterised, and examined. By employing SnCl2·2H2O and the transition metal precursors FeCl3 and NiCl2·6H2O with various Fe/Ni molar ratios, thermal annealing was carried out at a high temperature (700 °C). X-ray diffraction (XRD), UV–Visible spectroscopy, Photoluminescence (PL), FT-IR, and scanning electron microscopy (SEM) with energy dispersive X-ray techniques (EDX) were used to examine the materials' structural, chemical, optical, morphological, and anti-microbial capabilities. The average particle size of pure and co-doped SnO2 nanoparticles was determined to be around 52 nm and 15 nm, and SnO2 crystallites were observed to present tetragonal rutile structure with space group P42/mmm (No.136). Metal ions were replaced in the Sn lattice, as shown by Fe and Ni co-doped SnO2 nanoparticles. Pure and co-doped samples have capsule and sphere-like features in their SEM morphology. Using UV–visible diffuse reflectance spectroscopy, the optical property was examined, and it was observed that the band gaps for pure and co-doped SnO2 were 3.73 eV and 3.53 eV, respectively. The functional groups and incorporation of Fe and Ni in the prepared powder were also validated by FT-IR and EDX studies. By utilising the agar well diffusion technique and Nutrient agar, the antibacterial properties of pure, Ni-Fe co-doped SnO2 nanoparticles annealed at 700 °C were assessed. They were evaluated against various Gram-positive bacteria (Staphylococcus pheumoniae) and Gram-negative bacteria (Shigella dysenteria). The zone of incubation was found against the Gram +Ve and Gram −Ve bacterial strains.