RIBSE - Summer Research Institute for Biomedical Materials Science & Engineering

Robert Hard, Ph.D.

Pathology and Anatomical Sciences
University at Buffalo
315 Biomedical Research Building
Buffalo, NY
Tel: (716) 829-3521 VOICE
Email: Hard@buffalo.edu

The Hard group is focusing on using live cell microscopic imaging techniques to ascertain and quantify cellular phenomena, especially those dealing with respiratory epithelial cells by:

Determining cellular behaviors involved in re-epithelialization. We are developing methods for long term observation of living cultured respiratory epithelial cells undergoing wound healing. We have developed two modalities to reproducibly wound cultures. We are developing the use of microinjection and other methods to incorporate molecules labeled with caged or uncaged dyes into cells of known phenotype and then follow these cells during re-epithelialization. We are developing methods to track the cells and to quantify how different cell types participate in mitotic and motility aspects of re-epithelialization. With Drs. Hicks, Lwebuga-Mukasa, Gardella and Bright, we are developing methods to quantify the effects of cytokines and growth factors released from a resorbable laminated repair membrane (RLRM) on the rate and pattern of re-epithelialization.

Developing functional, demembranated, and reactivated models that include: isolated epithelial sheets, isolated ciliated cells, demembranated ciliary tufts, isolated ciliary axonemes and motor molecules associated with cilia. Each of these "model" systems allows one to study behavior at a different level of organization without the control mechanisms present in higher order models.

Developing new methods using high speed digital imaging to quantify ciliary beat frequency, ciliary waveform and ciliary coordination in the models listed in item II. The methods for quantifying coordination include Fourier analysis of deconvolved differential interference contrast high speed image sequences and the correlation of the beat frequency, phase and direction of each of the several hundred cilia per cell with fluorescent data designed to quantify how different proteins affect the coordination process.

With Dr. Sachs, testing and implementing a new xenon strobe light source for microscopy that provides high intensity flashes at up to 1000 Hz. Currently 4 flash lamps can be randomly fired, whose outputs are each passed through a distinct filter and then combined in a common fiber optic that is interfaced to the microscope. Therefore, multiple wavelengths are delivered at high intensity and at a high rate of speed and are of uniform intensity in the object plane. Even more remarkable, the output of each flash is individually monitored and adjusted to produce a constant intensity. Therefore, these features, multiple wavelengths at high intensity and flash rate that are uniform in space and time, allow for unparalleled image quantification.