Intact tissue is frozen in liquid nitrogen to be imaged by a cryo microtome. We use an imaging CryoMicrotome for 3D fluorescence of frozen lung tissue. This instrument is an automated image acquisition and analysis system consisting of hardware and software designed to collect fluorescence images of tissue sections. A motor-driven microtome sequentially sections frozen tissue while mercury filtered light excites fluorophores in the exposed surface of the tissue block. At each slice, a CCD camera records fluorescence images to be later analyzed for fluorescence distribution display. The microtome is housed in a freezer unit that maintains the sample at -40C during sample slicing and image acquisition. The resolution in the z direction of microtome cuts is 10 µm.
The cryoimager has been used to scan fluorescence signals from a variety of organs including lungs, hearts, kidneys, eyes, livers, and brains.
Schematic of Cryoimager
3D rendered fluorescence images of lung tissues:
Rat lung after ischemia reperfusion injury:
Rat lung after subjection to hyperoxia:
Rat eye at 3 weeks of age:
Histogram Comparison of Genetic Mutation in Kidneys:
Histogram Comparison of Genetic Mutation in Eyes:
Histogram Comparison of Kidneys Treated with Topicamide Compared to Vehicle Treated; Kidneys are diabetic due to injection with streptozotocin (STZ).
Histogram Comparison of Genetically Modified Kidneys; AK/+TSP1-/- serves as a model of severe diabetes, with TSP1-/- is its control.