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. Author manuscript; available in PMC: 2021 May 28.
Published in final edited form as: J Am Soc Mass Spectrom. 2020 Jul 24;31(12):2426–2436. doi: 10.1021/jasms.0c00119

Figure 1.

Figure 1.

Workflow of sample preparation and multimodal imaging. (1) The cornea is removed before fixation with 4% paraformaldehyde for 48 h at 4 °C. The iris and lens are removed before imaging. (2) The eye lacking cornea, iris, and lens mounted in jig to obtain OCT image. (3) The eye is placed in a dissection guide as described in the text to capture a belt with the optic nerve, macula, and temporal periphery. The belt is embedded in 2.6% CMC in a cryomold. The caps are discarded. (4) Cryosections at 12–14 μm throughout the entire eye are thaw-mounted on either glass or ITO slides. (5) ITO slides are imaged for autofluorescence (AF) before being coated with matrix via sublimation for acquisition of IMS data. (6) Highly accurate data registration is performed from IMS, postacquisition AF, pre AF, and H&E stained tissue.