SPECT projections are contaminated by scatter, resulting in reduced image contrast and quantitative errors. When tissue is present behind the source, some of the detected photons backscatter via this tissue. Particularly in dual-isotope SPECT and in combined emission-transmission SPECT, backscatter constitutes a major part of the down-scatter contamination in lower-energy windows. In this paper, the effects of backscatter material were investigated. Planar images of 99mTc and 201Tl line sources between varying numbers of Perspex slabs were analysed using the photopeak windows and various scatter windows. In the 99mTc photopeak window no significant change in total counts due to backscatter material was measured. In the 201Tl photopeak window an increase of about 10% in total counts was observed. In the scatter windows an even more explicit influence of backscatter material was measured. For instance, at a forward depth of 10 cm, total counts of a 99mTc source detected in the 72 keV window eventually doubled with increasing backscatter material, compared with the situation without backscatter material. The backscatter contribution plateaued when more than 5-10 cm of scatter material was placed behind the source. In conclusion, backscatter should be taken into account, particularly in model-based down-scatter correction methods in dual-isotope SPECT and combined emission-transmission SPECT.