Two-photon excitation 4Pi confocal microscope: Enhanced axial resolution microscope for biological research

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Abstract
The applicability of two photon excitation 4Pi confocal fluorescence microscopy to biological imaging is demonstrated. We show that 4Pi confocal microscopy in combination with a simple deconvolution algorithm allows axial localization and quantification with 0.14 μm resolution in a biological sample. The 4Pi‐confocal microscope extends the applicability of far field fluorescence microscopy to high resolution three‐dimensional imaging and quantification of subcellular structures.