ABSTRACT
Sub diffraction limited infrared absorption imaging of hemoglobin was performed by coupling IR optics with an atomic force microscope. Comparisons between the AFM topography and IR absorption images of micron sized hemoglobin features are presented, along with nanoscale IR spectroscopic analysis of the metalloprotein.
Subject(s)
Hemoglobins/chemistry , Image Processing, Computer-Assisted/methods , Microscopy, Atomic Force/methods , Nanostructures/chemistry , Spectroscopy, Fourier Transform Infrared/methods , Absorption , Hemoglobins/ultrastructure , Humans , Image Processing, Computer-Assisted/instrumentation , Lasers , Metalloproteins/chemistry , Metalloproteins/ultrastructure , Microscopy, Atomic Force/instrumentation , Nanostructures/ultrastructure , Reproducibility of Results , Sensitivity and Specificity , Spectroscopy, Fourier Transform Infrared/instrumentationABSTRACT
IR absorption spectroscopy of hemoglobin was performed using an infrared (IR) optical parametric oscillator laser and a commercial atomic force microscope (AFM) in a novel experimental arrangement based on the use of a bottom-up excitation alignment. This experimental approach enables detection of protein samples with resolution much higher than that of standard IR spectroscopy. Presented here are AFM-based IR absorption spectra of micron-sized hemoglobin features.
Subject(s)
Hemoglobins/analysis , Spectroscopy, Fourier Transform Infrared/methods , Hemoglobins/chemistry , Hemoglobins/metabolism , Lasers , Microscopy, Atomic Force/methods , Oscillometry , Reproducibility of Results , Sensitivity and Specificity , Spectroscopy, Fourier Transform Infrared/instrumentation , Time FactorsABSTRACT
Infrared absorption spectroscopy of lipid layers was performed by combining optics and scanning probe microscopy. This experimental approach enables sub-diffraction IR imaging with a spatial resolution on the nanometer scale of 1, 2-dioleoyl-sn-glycero-3-phosphocholine lipid layers.
