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. 2012 Oct 17;103(8):1691–1697. doi: 10.1016/j.bpj.2012.09.014

Figure 1.

Figure 1

Scheme of the line-scan confocal microscope. (a) An optical design of the line-scan confocal microscope. The new microscope was built by modifying a commercial inverted microscope (IX71, Olympus). For versatility of the microscope, three lasers—473-nm (Blues TM50, Cobalt), 532-nm (Compass215M, Coherent), and 640-nm (Cube640-100C, Coherent)—were used as light sources. CL1, a cylindrical lens (f = 40 mm, LJ1125L1-A, Thorlabs, Newton, NJ); CL2, a cylindrical lens (f = 50 mm, LJ1695L2-A, Thorlabs); CL3, a cylindrical lens (f = 250 mm, LJ1267L1-A, Thorlabs); L1, a spherical lens (f = 250 mm, LA1461-A, Thorlabs); L2, a spherical achromatic lens (f = 250 mm, LAO-250.0-50.0/075, Melles-Griot); L3, a spherical achromatic lens (f = 300 mm, LAO-300.0-50.0/075, Melles-Griot); L4 and L5, spherical achromatic lenses (f = 100 mm, LAO-100.0-50.0/075, Melles-Griot, Albuquerque, NM); L6, a spherical achromatic lens (LAO-350.0-40.0/Specialty HEBBAR coating for 415nm to 700nm, Melles Griot); PM, a polychroic mirror (z473/532/633rpc-xt, Chroma, Muenster, Germany); GM1 and GM2, galvanometric mirrors (VM1000+, General Scanning); OL, an objective lens (UPLSAPO100XO, or UPLSAPO60XW, Olympus); S, a slit (S30R or S40R, Thorlabs). (b) Side view of the sample plane. The illumination beam (green) is focused in a line shape on the sample plane. Only the fluorescence signal near the focal plane (yellow) can filter through a confocal slit and thus be detected. (c) Scheme of 2-D image generation. The illumination line on the sample plane (green) and the fluorescence image on a CCD camera (red) are simultaneously scanned once per filming cycle of the camera.