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Understanding why some leukemia cells undergo apoptosis upon venetoclax (VEN) treatment while others remain refractory is essential for improving therapeutic outcomes. Here, we integrate Raman microscopy, FT-IR imaging, and chemometrics to explore treatment-associated spectroscopic patterns linked to differential VEN response in Philadelphia-positive B-cell acute lymphoblastic leukemia (Ph+ B-ALL). Using two BCR-ABL1-positive cell lines with intrinsic differences in VEN sensitivity (BV-173, sensitive; SD-1, resistant), we combined classical biological readouts of apoptosis with multimodal vibrational profiling to characterize global biochemical changes induced by VEN. Sensitive BV-173 cells displayed spectroscopic changes consistent with apoptotic progression, reflected in coordinated alterations in nucleic acids, proteins, and lipids. In contrast, resistant SD-1 cells exhibited a distinct treatment-associated biochemical response profile consistent with nonapoptotic adaptation. Spectroscopic fingerprints enabled differentiation between apoptotic and nonapoptotic response patterns in the cell-line models studied. Our findings demonstrate that vibrational spectroscopy, integrated with chemometrics, may provide a complementary analytical framework for exploratory characterization of treatment-associated biochemical phenotypes in leukemia.