Real-time imaging reveals unique heterogeneous population features in insect cell cultures.

•Live cell imaging revealed heterogeneous populations in Sf9 insect cell cultures.•Live cell imaging revealed the presence of motile elongated cells that were resistant to baculovirus infection.•The migration speed of motile elongated cells was measured, and was comparable to that of melanoma cells.•Low or high dissolved oxygen induced the appearance of motile cells, suggesting that stress triggers their appearance.•A dual recombinant baculovirus with fluorescent proteins under promoters with different temporalities in the infection cycle allowed the identification of conditions that altered the infection cycle.