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). We present that the detectability of nano-sized cell membranes is enhanced to well below the optical diffraction limit since the membranes are found to act as thermal barriers. VIPPS offers a novel combination of chemical imaging and thermal diffusion characterization that paves the way towards label-free imaging of cell models and tissues as well as the study of intracellular heat dynamics.
Optical phase and birefringence signals occur in cells and thin, semi-transparent biomaterials. A dual-modality quantitative phase and polarization microscope was designed to study the interaction of cells with extracellular matrix networks and to relate optical pathlength and birefringence signals within structurally anisotropic biomaterial constructs. The design was based on an existing, custom-built digital holographic microscope, to which was added a polarization microscope utilizing liquid crystal variable retarders. Phase and birefringence channels were calibrated, and data was acquired sequentially from cell-seeded collagen hydrogels and electrofabricated chitosan membranes. Computed phase height and retardance from standard targets were accurate within 99.7% and 99.8%, respectively. Phase height and retardance channel background standard deviations were 35 nm and 0.6 nm, respectively. Human fibroblasts, visible in the phase channel, aligned with collagen network microstructure, with retardance and azimuth visible in the polarization channel. Electrofabricated chitosan membranes formed in 40 µm tall microfluidic channels possessed optical retardance ranging from 7 to 11 nm, and phase height from 37 to 39 µm. These results demonstrate co-registered dual-channel acquisition of phase and birefringence parameter maps from microstructurally-complex biospecimens using a novel imaging system combining digital holographic microscopy with voltage-controlled polarization microscopy.
- Publication Date:
- NSF-PAR ID:
- 10369214
- Journal Name:
- Biomedical Optics Express
- Volume:
- 13
- Issue:
- 2
- Page Range or eLocation-ID:
- Article No. 805
- ISSN:
- 2156-7085
- Publisher:
- Optical Society of America
- Sponsoring Org:
- National Science Foundation
More Like this
-
-
Abstract Electrospun biopolymer fibers are utilized in a wide variety of industries such as tissue engineering, sensors, drug delivery, membrane filtration, and protective membranes. The biopolymer chitosan, the partially
N ‐deacetylated derivative of chitin, which has been the focus of many studies, contains amine or hydroxyl functionalities that may be substituted with a number of chemistries such as carboxylate, benzene, or cyano groups. Modified chitosan solutions are often challenging to electrospin, as an entirely new set of solution and operating conditions must be developed for each modification. In this study, a facile post‐modification processing method for chitosan is introduced that circumvents the need to perform bulk modification prior to electrospinning, and therefore new spinning conditions. The chitosan mats were solution‐phase post‐processed by chemically functionalizing the mats with carboxylate, benzene and cyano groups. Scanning electron microscopy and Fourier‐transform infrared have been performed to determine fiber morphology retention and chemical interactions, respectively. Post‐modification retained the fibrous structure of the white‐colored, round and smooth mats with spectral changes indicating changes in the chitosan mat. Mean fiber diameters were 131 ± 75 nm (~31% smaller), 210 ± 81 nm (46% larger), and 85 ± 29 nm (~11% smaller) for carboxymethylchitosan, benzylidenechitosan, and cyanochitosan, respectively. -
In this paper, we discuss flat programmable multi-level diffractive lenses (PMDL) enabled by phase change materials working in the near-infrared and visible ranges. The high real part refractive index contrast (Δn ∼ 0.6) of Sb 2 S 3 between amorphous and crystalline states, and extremely low losses in the near-infrared, enable the PMDL to effectively shift the lens focus when the phase of the material is altered between its crystalline and amorphous states. In the visible band, although losses can become significant as the wavelength is reduced, the lenses can still provide good performance as a result of their relatively small thickness (∼ 1.5λ to 3λ). The PMDL consists of Sb 2 S 3 concentric rings with equal width and varying heights embedded in a glass substrate. The height of each concentric ring was optimized by a modified direct binary search algorithm. The proposed designs show the possibility of realizing programmable lenses at design wavelengths from the near-infrared (850 nm) up to the blue (450 nm) through engineering PMDLs with Sb 2 S 3 . Operation at these short wavelengths, to the best of our knowledge, has not been studied so far in reconfigurable lenses with phase-change materials. Therefore, ourmore »
-
Digital holographic microscopy provides the ability to observe throughout a volume that is large compared to its resolution without the need to actively refocus to capture the entire volume. This enables simultaneous observations of large numbers of small objects within such a volume. We have constructed a microscope that can observe a volume of 0.4 µm × 0.4 µm × 1.0 µm with submicrometer resolution (in xy) and 2 µm resolution (in z) for observation of microorganisms and minerals in liquid environments on Earth and on potential planetary missions. Because environmental samples are likely to contain mixtures of inorganics and microorganisms of comparable sizes near the resolution limit of the instrument, discrimination between living and non-living objects may be difficult. The active motion of motile organisms can be used to readily distinguish them from non-motile objects (live or inorganic), but additional methods are required to distinguish non-motile organisms and inorganic objects that are of comparable size but different composition and structure. We demonstrate the use of passive motion to make this discrimination by evaluating diffusion and buoyancy characteristics of cells, styrene beads, alumina particles, and gas-filled vesicles of micron scale in the field of view.
-
Over the last decade, the vector-apodizing phase plate (vAPP) coronagraph has been developed from concept to on-sky application in many high-contrast imaging systems on 8 m class telescopes. The vAPP is a geometric-phase patterned coronagraph that is inherently broadband, and its manufacturing is enabled only by direct-write technology for liquid-crystal patterns. The vAPP generates two coronagraphic point spread functions (PSFs) that cancel starlight on opposite sides of the PSF and have opposite circular polarization states. The efficiency, that is, the amount of light in these PSFs, depends on the retardance offset from a half-wave of the liquid-crystal retarder. Using different liquid-crystal recipes to tune the retardance, different vAPPs operate with high efficiencies (
) in the visible and thermal infrared (0.55 µm to 5 µm). Since 2015, seven vAPPs have been installed in a total of six different instruments, including Magellan/MagAO, Magellan/MagAO-X, Subaru/SCExAO, and LBT/LMIRcam. Using two integral field spectrographs installed on the latter two instruments, these vAPPs can provide low-resolution spectra ( ) between 1 µm and 5 µm. We review the design process, development, commissioning, on-sky performance, and first scientific results of all commissioned vAPPs. We report on the lessons learned and conclude with perspectives for future developmentsmore »
