Hcx pl apo 10x 0.4 na dry objective

We used a Leica TCS SP8 confocal laser scanning microscope, white light laser, Leica HCX PL APO 10X/0.4 NA dry objective (2.2 mm working distance) or Leica HC PL APO 40X/1.25 NA oil objective (0.24 mm working distance), and a SuperZ galvometric scanning stage. Using the 10X objective, one half of an immunostained and cleared core was scanned, the sample flipped over, and the other half scanned. 3D scanning was performed in a defined X/Y/Z (1.82/1.82/7.5 μm) volume with 10 frame averaging and bidirectional scanning using 488 nm excitation and 495–530 nm emission filter for DyLight488, 550 nm and 563–579 nm filters for DyLight550, 594 nm and 603–624 nm filter for DyLight594, 633 nm and 648–662 nm filter for DyLight633, 670 nm and 698–721 nm filter for DyLight680, 670 nm and 749–800 nm filter for DyLight755, and 550 nm excitation and 700–722 nm emission filters for LDS 751 nuclear staining.

Imaging was performed using a Leica TCS SP8 confocal laser scanning microscope, white light laser, Leica HCX PL APO 10X/0.4 NA dry objective (2.2 mm working distance) or Leica HC PL APO 40X/1.25 NA oil objective (0.24 mm working distance), and a SuperZ galvometric scanning stage. 3D scanning of macrosections was performed in a defined X/Y/Z (1.82/1.82/7.5 μm) volume with 10 frame averaging and bidirectional scanning using 488 nm excitation and 496-530 nm emission filter for DyLight488, 550 nm and 563-579 nm filters for DyLight550, 594 nm and 603-624 nm filter for DyLight594, 633 nm and 648-662 nm filter for DyLight633, and 670 nm excitation and 698-721 nm filter for DyLight680.