We show a platform that merges a microfluidic chip with lensless imaging for CD4 + T-lymphocyte counting at resource-limited settings. To capture CD4 + T lymphocytes, anti-CD4 antibody was immobilized on a microfluidic chip. The captured cells were detected by a charge coupled device (CCD) sensor using lensless shadow imaging techniques. Gray scale shadow images of captured cells on the chip (24 mm × 4 mm × 50 μm) were enumerated in three seconds using an automatic cell counting software. The device achieved 70.2 ± 6.5% capture efficiency, 88.8 ± 5.4% capture specificity for CD4 + T-lymphocytes, 96 ± 1.6% CCD efficiency, and 83.5 ± 2.4% overall platform performance (n = 3D 9 devices). This integrated platform has potential for point-of-care testing (POCT) to rapidly capture, image and count specific cell types from unprocessed whole blood.