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Histological and histochemical examinations of trace evidence were performed by lifting the specimens onto cellophane tape. To ensure adhesion of the samples during the staining, the surface of the tape was coated with 2% agarose after the sampling. These samples were ready for such direct stains as hematoxylin and eosin, benzidine, and periodic acid-Schiff to determine proteins and acid phosphatase. Histological analyses may be performed with the same range of smear preparations.

In forensic serology, A-B-H blood group substances have been successfully detected by mixed agglutination and absorption-elution techniques. By modifying the methods of Kouvarik et al [1] and Davidsohn [2], Ishiyama et al [3] recently developed a new technique for mixed cell agglutination reaction (MCAR) as follows: 1. Specimens were affixed onto the adhesive side of cellophane tape and then placed on a microscope slide. 2. Anti-A and anti-B agglutinins and Ulexanti-H lectin were used for sensitization. 3. The specimens were thoroughly washed to remove nonreactive agglutinins. 4. A corresponding indicator cell aliquot was added (sandwich method). 5. The slide was reversed and immersed in saline so that the nonreactive cells would dissociate spontaneously. 6. The results were read microscopically.

The electrochemical behavior and structural changes of several Nb2−xVxO5(x=0.2–1.0) were investigated in 1 M LiClO4-propylene carbonate solution as positive electrodes for rechargeable lithium batteries. Using the electrochemical measurements, X-ray diffractometry and X-ray photoelectron spectroscopy (ESCA), the possible electrode reaction with discharge and recharge is shown as follows: Nb2−xVxO5(x=0.2–1.0)+nLi++ne−αLinNb2−xVxO5. The discharge reaction consists of two main steps. In the first step, a ternary phase LinNb2−xVxO5(x=0.5–1.0) with a varying nvalue is formed, and in the second ternary phase LinNb2−xVxO5(x=0.5–1.0) with a varying nvalue is formed, and in the second ternary phases LinNb2−xVxO5with different constant nvalues may be formed. Good charge-discharge cycling behavior was obtained with the oxides having a higher xvalue (0.7–0.8), which showed a small crystal lattice change along the aand caxes, while keeping the original lattice arrangement during cycling.

For individual identification from semen, the deoxyribonucleic acid (DNA) fingerprint technique was used. In a blind trial, we succeeded in determining the semen donors among several volunteers comparing the DNA fingerprints of the blood and semen samples, respectively. Thereafter, we examined semen in a condom left beside a naked female dead body. The DNA fingerprint of the semen was recognized to be identical to that of the blood from a suspected man arrested later. This is the first report that the DNA fingerprint technique was practically used in a criminal investigation in Japan.

An improved method of the serological and morphological investigation of human hair is reported. The hair was firmly fixed onto a microscopic slide with cellophane tape and observed microscopically to confirm the cuticula images and the presence of the medulla. A piece of the hair containing the medulla was dissected, embedded in paraffin, and a cross section of this hair was prepared. By treating the sample with immunohistochemistry (biotin-antibiotin ABC technique), the blood type of the hair was confirmed definitively. Dust containing shaved beard can be examined in the same way.

A method for the demonstration of the topographical distribution of phenobarbital at the cellular level in various tissues was established. Mice that had been exposed to various doses of phenobarbital by intraperitoneal injection were killed, and their tissues were fixed with 0.1 Mphosphate buffer solution (pH 7.4) containing paraformaldehyde and glutaraldehyde. Thereafter, paraffin and frozen sections were made and stained by the indirect immunoperoxidase method using antisera obtained from commercial sources and used for the immunochemical assay of the blood level of phenobarbital in clinical medicine. A specific positive reaction was observed solely in testing the intoxicated tissues, and this reaction was inhibited when phenobarbital was added to the antisera. The minimal sensitivity of the positive reaction, which can be discerned by observing the stained slides macroscopically, was in the range of 10 mg/kg. Thus, the diagnosis of phenobarbital intoxication in the forensic autopsy can be made by immunohistochemistry. A positive reaction was found in various tissue cells, including nerve cells, myelin sheaths, glia cells, hepatocytes, cells of the alveolar and bronchial wall, epithelial cells of the distal part of the renal tubules, and so forth. Endothelial cells of the capillaries in all tissues gave a strong positive reaction. The immunocytochemical electron microscopy of the hepatocytes revealed that the positive reaction in the cytoplasm was located solely in the intraluminal space of the smooth endoplasmic reticulum. These results indicate some interesting aspects of the pharmacokinetics of phenobarbital in vivo. It is expected that the antisera, which are used widely for the assay of the blood concentration of various drugs (phenobarbital, amphetamines, morphine, and so forth), may be regarded as excellent reagents for immunocytochemistry. This clearly indicates that morphological evidence in toxicology, which had so far remained obscure, can be easily obtained by applying these antisera against various drugs.

A method for the demonstration of methamphetamine (MA) by immunocytochemistry was established. The tissues of intoxicated mice, administered various amounts of MA in single doses of from 0.01 to 1 mg of MA-HCl, were fixed in glutaraldehyde-containing fixatives. Cryostat and paraffin slices gave a positive reaction of MA localization by staining the brain, liver, kidney, lung, stomach, spleen, and so forth, with the aid of the indirect immunperoxydase technique. Those of animals administered a single dose of 0.1 mg or more (over 3 to 4 mg/kg—the usual dose of MA in acute intoxication death in forensic medicine), in particular, gave a strong strong reaction, so that the diagnosis of MA intoxication can be performed by macroscopic observation of stained slices. The histochemical diagnosis of MA intoxication in clinical toxicology and pathology might be regarded as a useful tool, especially in forensic pathology. The following cells gave a strong positive reaction: nerve cells and myelin sheaths, hepatocytes, epithelial cells of the distal part of the renal tubule and of the collecting tubule, alveolar and bronchial epithelial cells of the lung, chief and parietal cells of the gastric gland, capillaries of the renal glomerulus, macrophages in the blood and tissues, and striated muscle cells including cardiocytes. The morphological evidence of the pharmacodynamics and pharmacokinetics of MA can be determined at the cellular level by immunocytochemistry.

Human hair contains methamphetamine, amitriptyline, imipramine, nicotine, and their metabolites in some amount, which can be detected by routine toxicological methods. Sometimes, the level of drugs reaches over 100 μg/g. Animal experiments indicate that these drugs are found solely in sections of hair grown after administration of the drugs. The negative stage after the administration of drugs means that the hair section containing drugs has not come out of the hair follicle. Toxicological examination of the hairs may give some clue helping to identify the chronology of the intoxication.