Increasing contrast in water-embedded particles via time-gated mid-infrared photothermal microscopy.

The transient dynamics of photothermal signals provide interesting insights into material properties and heat diffusion. In a mid-infrared (mid-IR) photothermal microscope, the imaging contrast in a standard amplitude imaging can decrease due to thermal diffusion effects. It is shown that contrast varies for poly-methyl 2-methylpropenoate (PMMA) particles of different sizes when embedded in an absorbing medium of water (HO) based on levels of heat exchange under the water absorption resonance.

Time-Resolved Mid-Infrared Photothermal Microscopy for Imaging Water-Embedded Axon Bundles.

Few experimental tools exist for performing label-free imaging of biological samples in a water-rich environment due to the high infrared absorption of water, overlapping with major protein and lipid bands. A novel imaging modality based on time-resolved mid-infrared photothermal microscopy is introduced and applied to imaging axon bundles in a saline bath environment. Photothermally induced spatial gradients at the axon bundle membrane interfaces with saline and surrounding biological tissue are observed and temporally characterized by a high-speed boxcar detection system.

Thermal transport across membranes and the Kapitza length from photothermal microscopy.

An analytical model is presented for light scattering associated with heat transport near a cell membrane that divides a complex system into two topologically distinct half-spaces. Our analysis is motivated by experiments on vibrational photothermal microscopy which have not only demonstrated remarkably high contrast and resolution, but also are capable of providing label-free local information of heat transport in complex morphologies. In the first Born approximation, the derived Green's function leads to the reconstruction of a full 3D image with photothermal contrast obtained using both amplitude and phase detection of periodic excitations.

Erratum: Label-free imaging of fibroblast membrane interfaces and protein signatures with vibrational infrared photothermal and phase signals: publisher's note.

[This corrects the article on p. 303 in vol. 12, PMID: 33520386.

Label-free imaging of fibroblast membrane interfaces and protein signatures with vibrational infrared photothermal and phase signals.

Label-free vibrational imaging of biological samples has attracted significant interest due to its integration of structural and chemical information. Vibrational infrared photothermal amplitude and phase signal (VIPPS) imaging provide label-free chemical identification by targeting the characteristic resonances of biological compounds that are present in the mid-infrared fingerprint region (3 µm - 12 µm). High contrast imaging of subcellular features and chemical identification of protein secondary structures in unlabeled and labeled fibroblast cells embedded in a collagen-rich extracellular matrix is demonstrated by combining contrast from absorption signatures (amplitude signals) with sensitive detection of different heat properties (lock-in phase signals).

Phase-sensitive lock-in detection for high-contrast mid-infrared photothermal imaging with sub-diffraction limited resolution.

Imaging of the phase output of a lock-in amplifier in mid-infrared photothermal vibrational microscopy is demonstrated for the first time in combination with nonlinear demodulation. In general, thermal blurring and heat transport phenomena contribute to the resolution and sensitivity of mid-infrared photothermal imaging. For heterogeneous samples with multiple absorbing features, if imaged in a spectral regime of comparable absorption with their embedding medium, it is demonstrated that differentiation with high contrast is achieved in complementary imaging of the phase signal obtained from a lock-in amplifier compared to standard imaging of the photothermal amplitude signal.