There has been an increasing interest for observing fast biological phenomena such as cell movements in circulations and action potentials. The laser scanning confocal microscopy offers a good spatial resolution and optical sectioning ability to observe various in vivo animal models. We developed a high speed laser scanning confocal microscope capable of acquiring 512 by 512 pixel images at 200 fps (frames per second). We have incorporated a fast rotating polygonal scanning mirror with 128 facets for the X-axis scanner. In order to increase the throughput of the Y-axis scanner, we applied a bi-directional scanning method for vertical scanning. This made it possible to scan along the Y-axis two times during each scanner motion cycle. For the image acquisition, we used a custom photomultiplier tube amplifier with a broad frequency band. In addition, custom imaging software was written for the new microscope. In order to verify the acquisition speed of the developed confocal microscope, a resolution target moving at a series of constant speeds and a sedated mouse with slight movements due to heartbeats were observed. By comparing successive frames, the frame acquisition speeds were calculated.

We introduce an in vivo imaging flow cytometer, which provides fluorescence images simultaneously with quantitative information on the cell population of interest in a live animal. As fluorescent cells pass through the slit of light focused across a blood vessel, the excited fluorescence is confocally detected. This cell signal triggers a strobe beam and a high sensitivity CCD camera that captures a snapshot image of the cell as it moves down-stream from the slit. We demonstrate that the majority of signal peaks detected in the in vivo flow cytometer arise form individual cells. The instrument’s capability to image circulating T cells and measure their speed in the blood vessel in real time in vivo is demonstrated. The cell signal irradiance variation, clustering percentage, and potential applications in biology and medicine are discussed.

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We demonstrated the feasibility of blood cell assisted in vivo Particle Image Velocimetry using confocal microscopy. Blood flow of skin vessel in a mouse was non-invasively imaged in vivo using a confocal microscopy. The video-rate confocal microscope was used to monitor the motion of the blood cells in the capillary of a live mouse ear. The home-built confocal laser scanning microscopy allowed us to take images at the acquisition rate of 30 frames per second. The individual blood cells could be distinguished from other cells and the trajectory of the each cell could be followed in the sequential images. The acquired confocal images were used to get the velocity profile of the in vivo blood flow in conjunction with the Particle Image Velocimetry (PIV), without injecting any exogenous nano/micro particles into the mouse. We were able to measure the blood velocity up to a few hundreds µm/sec for various vessels in a live mouse. Because there is no need for the injection of the exogenous tracing particles, it is expected that we could apply the current technology to the study of human capillary blood stream.

Topical liquid formulations, dissolving microneedles (DMNs), and microscale needles composed of biodegradable materials have been widely used for the transdermal delivery of active compounds for skincare. However, transdermal active compound delivery by topical liquid formulation application is inhibited by skin barriers, and the skincare efficacy of DMNs is restricted by the low encapsulation capacity and incomplete insertion. In this study, topical serum application via a dissolvable micro-channeling system (DMCS) was used to enhance serum delivery through micro-channels embedded with DMNs. Transdermal serum delivery was evaluated after the topical-serum-only application and combinatorial serum application by assessing the intensity of allophycocyanin (APC) loaded with the serum in the porcine skin. APC intensity was significantly higher in the skin layer at a depth of 120–270 μm upon combinatorial serum application as compared to topical-serum-only application. In addition, the combinatorial serum application showed significantly improved efficacy in the clinical assessment of skin hydration, depigmentation, improvement of wrinkles, elasticity, dermal density, skin pores, and skin soothing without any safety issues compared to the serum-only application. The results indicate that combinatorial serum application with DMCS is a promising candidate for improving skincare treatments with optimal transdermal delivery of active compounds.

We experimented with two polymer materials with different ultraviolet (UV) wavelength absorption characteristics, which are commonly used in flexible devices, by applying an ultrashort-pulsed laser of a 355-nm UV wavelength for 10 ps. The laser parameters studied were pulse repetition rate, laser irradiation method, and laser power condition. Previous studies using polyethylene terephthalate (PET), which does not exhibit linear absorption at a UV wavelength, have focused on processing trends resulting in minimal collateral damage around the laser-induced ablation. However, our results showed a trend of accumulating such damage irrespective of the laser parameters. Meanwhile, polyimide (PI) exhibited a completely different behavior depending on the laser parameters. At low pulse repetition rates, minimal collateral damage was observed, whereas at high repetition rates, the morphology varied considerably. The electrical characteristics of the laser-processed materials were found to be correlated with the variations in morphology. In the case of PI, such variations in electrical resistance and morphology indicated that the material was carbonized. The findings of this study are expected to provide a useful reference when selecting parameters for the laser processing of similar polymer materials.