Primary bone marrow macrophages derived from mouse plated on a coverslip and imaged live. Internal Reflection Microscopy (IRM) shows direct apposition of the cells to coverslip with very little depth of field. Note the light gray uniform background extends under the cells' footprints demonstrating that the cells do not adhere tightly to the substrate. At the center of the image is a large polarized lamellipod of a crawling cell. The small spot below the cell is the tip of glass micropipette delivering CSF (colony stimulating factor) locally and the cell may be responding.
Live bone marrow macrophages plated in MatTek dish and imaged with a 60X N.A. 1.4 phase 3 Olympus objective on an IX71 microscope with a Cooke Sensicam QE camera and IPLab software running on a Dell Windows XP computer. The reflection microscopy was performed by removing the emission filter from a standard narrow pass rhodamine filter cube. Light was at approximately 570 nm and reflection leaked through the dichroic to allow for imaging.
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