Yuichi Komachi

Micro-optical fiber probe for use in an intravascular Raman endoscope

We believe that we have developed the narrowest optical-fiber Raman probe ever reported, 600 microm in total diameter, that can be inserted into coronary arteries. The selection of suitable optical fibers, filters, and a processing method is discussed. Custom-made filters attached to the front end of a probe eliminate the background Raman signals of the optical fiber itself. The experimental evaluation of various optical fibers is carried out for the selection of suitable fibers. Measurement of the Raman spectra of an atherosclerotic lesion of a rabbit artery in vitro demonstrates the excellent performance of the micro-Raman probe.

Microlenses and microlens arrays formed on a glass plate by use of a CO 2 laser

Microlenses and microlens arrays were formed directly on a surface of a glass plate by use of a CO(2) laser. When the surface of a glass plate is heated locally to a working point of the glass material by use of a focused CO(2) laser beam, it tends to become a hyperboloid owing to surface tension, which results in a microlens. A profile of the microlens was measured with an ultrahigh accurate three-dimensional profilometer (Model UA3P, Matsusita Electric Industrial Company Ltd.) that utilizes a specially designed atomic force microscope. An intensity profile and a spot diameter at the focus of the microlens were measured with a microscope and a CCD system utilizing a He-Ne laser as a light source. The focused spot FWHM diameter of 1.35 mum was obtained, and the modulation transfer function was derived from the spot profile. Microlens arrays were also fabricated and characterized.

Raman probe using a single hollow waveguide

A simple Raman probe was realized using a single flexible hollow waveguide (HW). A HW coated with a silver film, which had reasonable transmission and little optical background noise, was used as a bidirectional transmission fiber for both the excitation and collection of Raman scattered light. The HW itself generated no Raman scattering or fluorescence noise during transmission. A complex filtering system at the end of the waveguide was thus unnecessary. In addition, the measured Raman spectra showed better signal-to-noise ratios than a conventional Raman fiber probe. The HWs suitability as a Raman fiber probe was also demonstrated.

In Vivo Raman Study of the Living Rat Esophagus and Stomach Using a Micro-Raman Probe under an Endoscope:

A small endoscope system equipped with a micro Raman probe is developed for in vivo Raman measurements in living rats. The measurements are done under anesthesia and artificial respiration to minimize the impact on the rats. Raman spectra of living rat esophagus and stomach are successfully measured. Our results suggest that the Raman spectra reflect subsurface tissue structure that cannot be distinguished in the endoscope image. After the experiments, rats recover without any aftereffects. It is verified that the Raman measurement using the present system is safe and noninvasive for rats.

Improvement and analysis of a micro Raman probe

A micro Raman probe (MRP) with a 600 microm diameter, which we previously reported as the narrowest achieved to date, was further improved by introducing high-quality optical filters and a collecting lens at the tip. We fabricated the MRP with a high collection efficiency, a wider collection wavelength, and a high signal-to-noise ratio. We compared two types of probes: one with a lens-tipped end and one with a flat tip. We experimentally tested the performance of these MRPs to evaluate the detection properties defined by parameters such as the optical purity against inherent Raman background noise due to optical fibers, the sensitivity, and the viewing area. Finally, we demonstrated their effectiveness in measurements of standard Raman samples and applied them to measurements of plastic and human skin samples in situ.

Intravascular Raman spectroscopic catheter for molecular diagnosis of atherosclerotic coronary disease

An intravascular catheter for Raman spectroscopic detection and analysis of coronary atherosclerotic disease has been developed. The catheter, having an outer diameter of 2 mm, consisted of a side-view-type micro-Raman probe, an imaging fiber bundle, a working channel (injection drain), and a balloon. By inflating the balloon, the probe was brought close to the inner wall of a modeled blood flow system and detected a phantom target buried in the wall. Results obtained demonstrate the possibility of using the spectroscopic catheter for molecular diagnosis of coronary lesions.

Fluorescence-suppressed raman technique for quantitative analysis of protein solution using a micro-raman probe, the shifted excitation method, and partial least squares regression analysis

A practical Raman analyzing technique with suppression of the strong fluorescent background in order to obtain quantitative information is proposed in the present study. The technique is based on the shifted excitation method and partial least squares regression (PLSR) analysis. The Raman system consists of a single Raman spectrometer, a background-free electrically tunable Ti: Sapphire laser (BF-ETL), and a micro-Raman probe (MRP). The system allows one to obtain reliable shifted excitation Raman spectra with a simple operation. The PLSR analysis successfully provides quantitative information from the obtained spectra with the suppression of random noise including photon shot noise. The present study demonstrates that the technique is effective for extracting quantitative information concealed behind a fluorescent background that is more than 200 times stronger than the Raman signal.

Fiber-Optic Raman Probes for Biomedical and Pharmaceutical Applications

This chapter reviews the development of optical fiber probe Raman systems and their applications in life science and pharmaceutical studies. Especially, it is focused on miniaturized Raman probes which open new era in the spectroscopy of the life forms. The chapter also introduces the important optical properties of conventional optical fibers to use for Raman probes, as well as new types of optical fiber and devices, such as hollow optical fibers and photonic crystal fibers.

Detection of ER stress in vivo by Raman spectroscopy

The endoplasmic reticulum (ER) is an organelle in which most membrane and secretory proteins are synthesized. If these proteins are not folded correctly, unfolded proteins accumulate in the ER lumen, causing a cellular situation known as ER stress. Recently, many studies on the relationship between ER stress and diseases have been reported. Thus, studies of ER stress in vivo should yield information that is useful in pathology. Model mice have been developed as a powerful tool to visualize ER stress in vivo, but this approach depends on transgenic technology. Here, we report on a method of detecting ER stress in vivo by Raman spectroscopy. Our experiments revealed that two specific Raman bands were reduced in both cultured cells and animal tissues in an ER stress dependent manner. This suggests that Raman spectroscopy could be a useful tool to detect ER stress in vivo without transgenic technology.

An optical biopsy system with miniaturized Raman and spectral imaging probes: in vivo animal and ex vivo clinical application studies

An optical biopsy system which equips miniaturized Raman probes, a miniaturized endoscope and a fluorescent image probe has been developed for in vivo studies of live experimental animals. The present report describes basic optical properties of the system and its application studies for in vivo cancer model animals and ex vivo human cancer tissues. It was developed two types of miniaturized Raman probes, micro Raman probe (MRP) made of optical fibers and ball lens hollow optical fiber Raman probe (BHRP) made of single hollow optical fiber (HOF) with a ball lens. The former has rather large working distance (WD), up to one millimeter. The latter has small WD (~300μm) which depends on the focal length of the ball lens. Use of multiple probes with different WD allows one to obtain detailed information of subsurface tissues in the totally noninvasive manner. The probe is enough narrow to be inserted into a biopsy needle (~19G), for observations of the lesion at deeper inside bodies. The miniaturized endoscope has been applied to observe progression of a stomach cancer in the same rat lesion. It was succeeded to visualize structure of non-stained cancer tissue in live model animals by the fluorescent image technique. The system was also applied to ex vivo studies of human breast and stomach cancers.