Analysis of molecular orientation in polymeric spherulite using polarized micro attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopic imaging.

Micro ATR-FTIR spectroscopic imaging enables the visualization of two-dimensional chemical distribution at a higher spatial resolution than macro-transmission FTIR imaging approach. In this study, micro ATR-FTIR imaging was applied for analysis of a specific morphology in a spherulite of poly(3-hydroxybutyrate) (PHB). The PHB spherulites crystallized at an isothermal condition, showed the fine band structure due to the twisting lamellar crystals during the spherulite growth under the polarized optical microscope (POM). In addition, the band structure observed in the PHB spherulite was the double band pattern in which the higher and lower birefringence banded areas alternatively appear due to the three-dimensional orientation of crystallographic axes and the biaxial refractive index ellipsoid of PHB crystalline structure. Micro ATR-FTIR spectroscopic imaging was employed for detecting the double band structure in the PHB spherulite. However, the obtained spectral images did not indicate any band structures. To detect the difference of molecular orientation among the double band structures, the micro ATR-FTIR imaging was performed with a linear polarizer at four different angles. The mean values of absorbance in each measured area changed depending on the polarizer angle. The in-plane molecular orientation to the tangential direction of spherulite, caused by the dependence of the average absorbance on the polarizer angles, was determined by the position of measured area in the spherulite and the linear dicroism of each of the spectral band used. To visualize the small difference of molecular orientation in the double band structure, micro ATR-FTIR images of the dichroic differences at three spectral bands were calculated from two different sets of polarizer angles. The micro ATR-FTIR images representing the dichroic differences displayed their corresponding distributions among three spectral bands. The complementary distributions of the dichroic difference were caused by the crystallographic orientation of b- and c-axes and were successfully visualized to reveal the pattern with the features less than 10 μm in size. The results achieved in this study were due to two advantages of the polarized micro ATR-FTIR imaging: the high spatial resolution of micro ATR-FTIR imaging technique, and the high sensitivity of polarization measurements. Thus, this work demonstrates the power of this spectroscopic approach for such analytical investigation.