Low-temperature (9–12 °C) pulsed electric field (PEF) was investigated in milk before cream separation at different intensities (9–27 kV/cm, 66 μs, 16–28 kJ/L) regarding its potential to render processing more sustainable, retain a high physico-chemical quality, enhance functional properties, and gently modify the structure of the milk fat globule membrane (MFGM). Cream volume per L milk were most efficiently increased by 31 % at the lowest PEF intensity in comparison to untreated milk and cream (P < 0.05). Untreated and PEF-treated milk and obtained cream were assessed with compositional (fat, protein, casein, lactose, and total solids content) and particle size distribution analyses, showing no significant differences (P ≥ 0.05) and, thus, indicating retention of ‘native-like’ product quality. Overrun and stability of cream, whipped for 20 and 60 s at 15000 rpm using a high-shear mixer, were improved most notably by the lowest and the highest PEF intensities, achieving up to 69 % enlarged overrun and up to 22 % higher stability, respectively (P < 0.05), than in untreated whipped cream. Protein component analyses for milk and cream were carried out by sodium dodecylsulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Noticeable differences between untreated and PEF-treated milk were not observed, but the SDS-PAGE results for cream showed noticeably different bands for some of the protein components, indicating structural changes in MFGM-, whey-, and phospho-proteins due to PEF and/or separator processing effects. More intense bands of xanthine oxidase, xanthine dehydrogenase, butyrophilin, bovine serum albumine, adipophilin (ADPH), and glycoproteins PAS6/7 were observed specifically at 21 kV/cm. Gentle electroporation of both MFGM layers by PEF was determined based on the changes in MFGM monolayer components, such as ADPH and PAS 6/7, exhibiting intensified bands. PEF intensity-dependent impact on the structure of MFGM and casein, leading to a reconfiguration of the cream matrix due to different structuring interactions among proteins, among milk fat globules, and between fat and protein components, was suggested. Overall, low-temperature PEF applied at different intensities showed great potential for gentle, efficient, and functional properties-tailored dairy processing and may also enable effective extraction of highly bioactive ingredients from dairy sources.