Scanning electron microscopic analysis of pancreatic tissue in alcoholic and tropical chronic pancreatitis.

INTRODUCTION: Chronic Pancreatitis (CP) is a heterogenous disease with alcoholic chronic pancreatitis (ACP) dominating in the West, and idiopathic or tropical chronic pancreatitis (TCP) in the tropics. The aim of this study is to assess the feasibility of using a scanning electron microscope (SEM) to analyze the ultra-structural changes in alcoholic and tropical subtypes of CP. METHODS: Chronic pancreatitis tissue samples were taken from the biopsy samples of 16 patients (seven ACP and nine TCP) who underwent drainage procedures for CP. These samples were subjected to SEM analysis and findings of normal pancreas were compared with those of CP for appreciating differences in their architectural changes. RESULTS: Normal architecture of pancreas could be observed as lobules of parenchyma, ductal system and definite loci of Islets of Langerhans (IOL). CP samples showed loss of architecture in the form of severe fibrosis and calcifications. In ACP, the fibrosis was predominantly seen towards the periphery of the gland sparing the periductal areas. These fibres were strangulating and damaging the parenchyma. Crystals were seen over these fibres. In TCP, fibrosis was moderate and uniform throughout the parenchyma. Moreover the crystals were larger and intraluminal. Total fatty replacement of parenchyma was a striking feature in TCP, seen exclusively in diabetics with gross atrophy of IOL. CONCLUSION: SEM gives the real-life pictures of fibrosis, fatty change, ductal changes, calcifications and thus the actual extent of damage in CP better than the ordinary light microscopy.

Problem in analyzing cystine stones using FTIR spectroscopy.

Cystine stones are produced by an inherited disorder of the transport of amino acid cystine that results in excess of cystine in the urine (cystinuria). Cystine calculi in urinary tract present a significant problem in patients. We have recorded that cystine calculi are very uncommon in our region. Cystine crystals are unusually identified in the urinary deposits. The problem of recognizing cystine by FTIR as a component in mixture of stones is significant. The problem is compounded by the similarity of wavelengths of cystine with that of whewellite and uric acid. The objective of this paper is to elucidate the problems of identifying cystine in stone analysis and identifying a solution to get over this deficiency. Out of 1,300 urinary stones analysed by ordinary wet chemical methods and infrared spectroscopy, 30 stone samples, which were reported to have cystine peaks in significant numbers, were selected. These samples were powdered, mixed with potassium bromide, pelletized and taken up for FTIR analysis. The wavelength patterns were scrutinized by comparing with the peaks obtained by the reference standards of cystine. Spectra were also obtained from pure cystine. Comparison of spectra with those of whewellite and uric acid was performed. Then the samples were taken for Scanning electron microscopy with elemental distribution analysis X-ray (SEM-EDAX). The samples were made conductive by gold sputtering and were fed into JEOL JSM 35 C SEM machine. Morphology was recorded by taking photographs. Further elemental distribution analysis (EDAX) was carried out to identify the elemental composition. Of the 30 samples taken up for FTIR analysis, all showed spectra identifiable with the reference peaks for cystine. However, when these peaks were compared with those of whewellite and uric acid, all the stone samples showed duplication of peaks for whewellite and uric acid and whewellite. The pure cystine spectra showed identifiable peaks are in the range of 3026, 1618.28, 1485, 846.75 cm(-1), etc. (from the standard spectrum of pure cystine). All the analysis findings were correlated with EDAX findings. On doing EDAX, we could separately find out the components present in a mixture. Three stones contained elemental pattern to fit with those of cystine. Even though it is difficult to find out the presence of cystine molecule in FTIR, it is possible to recognize it through EDAX and will be possible to confirm the presence of cystine in mixed urinary stones.