Cryoplane Microscopy

Drexel University, Philadelphia PA

Investigators

Abstract

This award supports development of a novel optical imaging device, the cryoplane microscope (CM). In the automated device will provide serial images of biological materials at a range of magnification from micro- to macro-scale. In the CM, a frozen specimen is cut on a cryostat and the exposed blockface surface imaged. By shaving sections off the block, deeper tissue levels are revealed. Successive images will be in register by virtue of the instrument's design; the resulting series of images will yield an aligned 3D dataset. The proposed instrument will support incident light brightfield and epifluorescence. It will support resolution as high as 0.7 microns and, due to the opacity of ice, the acquired images will be uncompromised by out-of-plane contamination. Fields-of-view, as large as 5 cm by 3 cm and as deep as 6 cm, will be accommodated. The instrument will not be limited to dissected material; whole small organisms can be imaged. Among potential uses are the mapping of gene expression, tracing of connectivity in the nervous system, and characterization of genetically manipulated specimens. Key to the utility of this technology is the high-throughput that will be achieved. Complex image acquisition protocols involving changes in resolution, excitation and emission filters, image integration time, illumination intensity, and field-of-view will be supported in both automated and supervised modes. All these operations can be directed remotely over the internet using software to be developed as part of the project. Typically, the 3D multispectral high-resolution datasets that the instrument will generate will be large enough that distribution of entire datasets via the internet would be difficult. Thus the visualization and image processing software to be developed will permit remote viewing of images, and will permit navigation through the volume to be guided remotely and at high-speed. Thus the software tools will facilitate potential use of the instrument and of data by other researchers, and by students in a classroom or other remote setting.