Table 2 Imaging modalities.
From: Deep tissue analysis of distal aqueous drainage structures and contractile features
Modality | Acronym | Color | Example | Represents |
|---|---|---|---|---|
1. Two-photon excitation fluorescence: | TPEF | Â | Â | Â |
a. Endogenous fluorophores | Â | Red | td-Tomato | Plasma membrane |
| Â | Â | Â | VE-cadherin-td-Tomato | Endothelium |
| Â | Â | Green | GFP | Cytoplasm |
| Â | Â | Â | Prox1-GFP | Endothelium of distal tract or lymphatics |
b. Epifluorescence | Â | Red | CD31 (aka PECAM1) | Endothelium |
| Â | Â | Â | F-actin (Alexa- 568-Phalloidin) | Contractile cytoskeleton |
| Â | Â | Â | LYVE-1 | Lymphatic endothelium |
| Â | Â | Â | VEGFR2 | Endothelium |
c. Intravital fluorescence | Â | Green | Hoechst 33342 | Nucleus |
| Â | Â | Â | siGLO | Nucleus |
d. Autofluorescence | TPEF AF | Green | Elastin and fibrillar collagen | Elastin and fibrillar collagen (e.g. types I and III) |
2. Second harmonic generation | SHG | Cyan | Fibrillar collagen | Fibrillar collagen (e.g. types I and III) ex: 850 nm; em: 425 nm |
3. Isosurface mapping | ||||
a. Signal | Â | Cyan | Collagen SHG | Fibrillar collagen (e.g. types I and III) |
| Â | Â | Green | siGLO | Trabecular meshwork |
| Â | Â | Red | VE-cadherin-td-Tomato | Blood vessel endothelium |
b. Signal voids |  | Yellow | Schlemm’s canal lumen | Distal drainage tract |
| Â | Â | Â | Collector channel lumen | Distal drainage tract |
| Â | Â | Â | Intrascleral plexus lumen | Distal drainage tract |
4. Immunohistochemistry | IHC | Red | F-actin | Contractility marker |
(by one-photon, confocal microscopy) | Â | Green | alpha-smooth muscle actin | Contractility marker with smooth muscle identity |
| Â | Â | Â | Caldemson | Contractility marker with smooth muscle identity |
| Â | Â | Â | Calponin | Contractility marker with smooth muscle identity |
