The present standard of blood cell analysis is an invasive procedure requiring the extraction of patient’s blood, followed by ex-vivo analysis using a flow cytometer or a hemocytometer. We are developing a noninvasive optical technique that alleviates the need for blood extraction. For in-vivo blood analysis we need a high speed, high resolution and high contrast label-free imaging technique. In this proceeding report, we reported a label-free method based on differential epi-detection of forward scattered light, a method inspired by Jerome Mertz's oblique back-illumination microscopy (OBM) (Ford et al, Nat. Meth. 9(12) 2012). The differential epi-detection of forward light gives phase contrast image at diffraction-limited resolution. Unlike reflection confocal microscopy (RCM), which detects only sharp refractive index variation and suffers from speckle noise, this technique is suitable for detection of subtle variation of refractive index in biological tissue and it provides the shape and the size of cells. A custom built high speed electronic detection circuit board produces a real-time differential signal which yields image contrast based on phase gradient in the sample. We recorded blood flow in-vivo at 17.2k lines per second in line scan mode, or 30 frames per second (full frame), or 120 frame per second (quarter frame) in frame scan mode. The image contrast and speed of line scan data recording show the potential of the system for noninvasive blood cell analysis.
Photothermal heterodyne imaging has demonstrated a high sensitivity of seeing single metallic nanoparticles of diameter
down to 5 nm. However, rare attention has been paid to the phase of the photothermal heterodyne signal relative to that
of the modulated pump beam. We show that the phase of the photothermal heterodyne signal from semiconducting
nanomaterials such as silicon and germanium nanowires is around 0 degree, while that from metallic nanomaterials such
as silver and gold nanoparticles (NPs) is around 180 degrees. Using this property we have been able to distinguish gold
seeds from germanium nanowire (GeNW) body in a label-free and contact-free manner. A theoretical model based on
light scattering by a fluctuating dielectric material was used to explain the origin of different phases in the photothermal
heterodyne signal.
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