Refractive-index-based calorimetric studies of RNAse T1 unfolding in small volumes using microinterferometric backscatter

Michael P. Houlne; Darren S. Hubbard; George I. Makhatadze; Darryl J. Bornhop

doi:10.1117/12.273610

2 May 1997 Refractive-index-based calorimetric studies of RNAse T1 unfolding in small volumes using microinterferometric backscatter

Michael P. Houlne, Darren S. Hubbard, George I. Makhatadze, Darryl J. Bornhop

Author Affiliations +

Proceedings Volume 2982, Optical Diagnostics of Biological Fluids and Advanced Techniques in Analytical Cytology; (1997) https://doi.org/10.1117/12.273610
Event: BiOS '97, Part of Photonics West, 1997, San Jose, CA, United States

Abstract

Micro-interferometry, a novel technique developed by the authors, employs a linearly polarized laser, a fused silica capillary tube housing for the sample and a charged coupled device as a detector. A back scattered interference pattern, observed as a high contrast fringes, is produced when the laser is directed onto the capillary containing the sample. The positional change of the fringe pattern is a function of the refractive index of the media in the capillary. In the present work, the RNA enzyme RNase T1 is heated in the sample cell over a temperature range of 30 degrees C to 60 degrees C. Over this temperature range the molecule unfolds form the quaternary to the tertiary structure. This structure change is manifested as a refractive index change and is observed by monitoring the fringe position while ramping the cell temperature in a controlled fashion. From the refractive index response over the temperature range, the Gibbs free energy associated with unfolding is calculated. The authors show milli-degree temperature stability with a 0.1 micro-liter probe volume, thus demonstrating the application of this device in micro- calorimetric investigations.

Citation Download Citation

Michael P. Houlne, Darren S. Hubbard, George I. Makhatadze, and Darryl J. Bornhop "Refractive-index-based calorimetric studies of RNAse T1 unfolding in small volumes using microinterferometric backscatter", Proc. SPIE 2982, Optical Diagnostics of Biological Fluids and Advanced Techniques in Analytical Cytology, (2 May 1997); https://doi.org/10.1117/12.273610

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