Call number: LD2668 .T4 1942 W6 / Master of Science

Accurate calculations of the axial chromatic aberration coefficients of geometrically symmetric three-cylinder tripotential electrostatic lenses are presented for two different center electrode lengths. This is an extension of the first-order properties and the third-order spherical aberration coefficients published by Harting and Read.

The theoretical basis for using electron mirrors as correctors of chromatic aberration is presented and an experimental verification of correction of chromatic aberration is demonstrated. A hyperbolic electrostatic electron mirror operating in its converging range and at unity magnification was used as a corrector. A novel separating system with deflections taking place at image planes was developed to implement the mirror without impairing the resolution. Correction was demonstrated in an electron optical probe system. The chromatic aberration was measured by means of the shadows cast by a fine mesh placed near the final image. The experimental method and equipment are described. The experiment serves as a verification of the theory as well as a successful test of the method of separating the electron beams traveling to and from the mirror.

Indiana University-Purdue University Indianapolis (IUPUI) / Introduction: Multiphoton fluorescence excitation microscopy (MPM) is an invaluable tool for studying processes in tissue in live animals by enabling biologists to view tissues up to hundreds of microns in depth. Unfortunately, imaging depth in MPM is limited to less than a millimeter in tissue due to spherical aberration, light scattering, and light absorption. Spherical aberration is caused by refractive index mismatch between the objective immersion medium and sample. Refractive index heterogeneities within the sample cause light scattering. We investigate the effects of refractive index mismatch on imaging depth in MPM. Methods: The effects of spherical aberration on signal attenuation and resolution degradation with depth are characterized with minimal light absorption and scattering using sub-resolution microspheres mounted in test sample of agarose with varied refractive index. The effects of light scattering on signal attenuation and resolution degradation with depth are characterized using sub-resolution microspheres in kidney tissue samples mounted in optical clearing media to alter the refractive index heterogeneities within the tissue. Results: The studies demonstrate that signal levels and axial resolution both rapidly decline with depth into refractive index mismatched samples. Interestingly, studies of optical clearing with a water immersion objective show that reducing scattering increases reach even when it increases refractive index mismatch degrading axial resolution. Scattering, in the absence of spherical aberration, does not degrade axial resolution. The largest improvements in imaging depth are obtained when both scattering and refractive index mismatch are reduced. Conclusions: Spherical aberration, caused by refractive index mismatch between the immersion media and sample, and scattering, caused by refractive index heterogeneity within the sample, both cause signal to rapidly attenuate with depth in MPM. Scattering, however, seems to be the predominant cause of signal attenuation with depth in kidney tissue. Kenneth W. Dunn, Ph.D., Chair