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PubMed1980

[Nomarski interference microscopy: use of combined contrast uranyl acetate-PTA for in vitro observation of monolayer cultures].

In this paper is reported a technique for the fixation and staining of cellular monolayers in vitro for observation by the interferential microscope according to Nomarski. This method includes the use of glutaraldehyde at 2.5% in phosphate buffer 0.1 M pH 7.5 and the sequential use of Uranil acetate (0.9% in absolute alcohol) and phosphtungstic acid (1% in absolute alcohol). This study has been conducted on 3 cellular lines: PC12 (rat pheocromocytoma), R.P.C. (rat pineal cells), primary culture obtained from a human carcinoma of the uterus.

Acetates↗
PubMedDate not supplied

Comparison of buccal and nasal epithelial cells using a new cell development index and quantitative interference microscopy.

Cell area, cell dry mass, and three summary indices for tissue maturation were obtained for human buccal and nasal cells viewed in smears (five donors). The means for cell area and dry mass in the buccal smears were 3816 X 10(-8) cm2 and 2294 pg (1 pg=1 X 10(-12) g), respectively. The correlation coefficient for means of cell area and dry mass was r=0.03. The means for the three maturation indices used to characterize the cells viewed in buccal smears were: Cell Development Index (CDI), 486; Maturation Index (MI) 50.6; and Karyopyknotic Index (KPI), 0.028. The correlation coefficients for means were: CDI vs. MI, r=0.88; CDI vs. KPI, r=0.71; and MI vs. KPI, r=0.73. In the buccal smears, the cell types found most frequently were intermediate with round or oval nuclei, which altogether averaged about 90% of the total smear composition, while superficial cells (pyknotic ghost, and anucleate) comprised less than four per cent. In nasal smears, the means for cell area and dry mass were 867 X 10(-8) cm2 and 623 pg, respectively. The correlation coefficient for means of cell area and dry mass was r=0.74. The means for the maturation indices were: CDI, 816; MI, 91.2; and KPI, 0.79. The correlation coefficients for means were CDI vs. MI, r=0.93; CDI vs. KPI, r=0.97; and MI vs. KPI, r=0.85. In nasal smears, the anucleate cell type was found most frequently (about 75%) while intermediate cells with round or oval nuclei comprised approximately 13% of the total smear composition. When donor means for cell area and dry mass of buccal cells were combined for study, they were highly correlated (r=0.88). Similarly, when the maturation indices for buccal and nasal cells were combined for study, they were highly correlated: CDI vs. MI, r=0.99; CDI vs. KPI, r=0.99; and MI vs. KPI, r=0.99. Of the three indices, we believe that the CDI is the index best suited for use in studies of cell development because it is based on an inferred sequence of cellular and nuclear changes occurring during the maturation of epithelial cells. It provides a more detailed description of the cells observed in the smears. It also correlates highly in KPI and MI used in clinical and research efforts.

Cell Nucleus↗
PubMed1975

Development with age of human articular cartilage surface structure. A survey by interference microscopy of the lateral femoral condyle.

The weight-bearing surfaces of the lateral femoral condyles taken from twenty normal human cadavers aged 0-47 years have been examined by reflected light interference microscopy (RLIM) and by scanning electron microscope (SEM). The surfaces appeared normal by naked eye examination. The presence of both 200-400 mum diameter secondary undulations and small ovoid 20-45 mum diameter tertiary hollows was confirmed in all specimens using both techniques. Measurements by RLIM showed that the tertiary hollows increase significantly in depth and diameter with increasing age. A further order of quaternary surface irregularities was shown. Small irregular ridges, 130-275 nm deep and 1-4 mum diameter, were found with increasing frequency on specimens obtained from persons aged 21 years. These quaternary irregularities are thought to be due to exposure of superficial fibre bundles after the loss, with age, of surface ground substance.

Adolescent↗
PubMed1993

Splenic function in inflammatory bowel disease: assessment by differential interference microscopy and splenic ultrasound.

The role of the spleen in patients with ulcerative colitis and Crohn's disease was assessed by counting pitted erythrocytes with differential interference microscopy and by splenic ultrasound. The findings were compared with those from age- and sex-matched controls and a group of splenectomized patients. The incidence of hyposplenism was lower than previously reported, being found in six of 29 patients with ulcerative colitis (of whom five had relapsed pancolitis and one had quiescent pancolitis) and two of 21 patients with Crohn's disease. Pitted erythrocyte counts were significantly higher in patients with ulcerative colitis compared with age- and sex-matched controls (p < 0.001), but there was no difference in counts between patients with Crohn's disease and controls. Patients with relapsed but not quiescent ulcerative colitis had significantly smaller spleens than controls. Patients with the highest pitted erythrocyte counts had the smallest spleens on ultrasound. More effective medical and surgical therapy may account for the lower incidence of functional hyposplenism observed in this study. Nonetheless, patients may show marked hyposplenism and remain at risk of overwhelming infection and operative complications. Differential interference microscopy is a simple technique that can be used to assess splenic function in patients thought to be susceptible to infection.

Adult↗
PubMed2005

Interference microscopy under double-wavelet analysis: a new approach to studying cell dynamics.

This Letter combines a novel experimental approach to the study of intracellular processes with a newly developed technique for multimode time-series analysis. Experiments are performed on isolated pond snail (Lymnaea stagnalis) neurons. Local variations in the cellular refractive index as detected by laser interference microscopy are related to the processes in the cell. A wavelet analysis shows the presence of several identifiable modes in the membrane and intracellular dynamics, and a double-wavelet analysis reveals nonlinear interactions between the regulatory processes in the form of mutual frequency and amplitude modulations.

Animals↗