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Biomedical subjects

A Manivannan

Publications and source records attributed to A Manivannan.

18 recordsLinked to original sources

The scanning laser ophthalmoscope--a review of its role in bioscience and medicine.

The scanning laser ophthalmoscope (SLO) offers the potential for retinal imaging that is complementary both to that of the fundus camera and also the newly developing technique of optical coherence tomography (OCT). It has the ability to produce rapid images at low light levels using light of specific wavelengths. This permits temporal studies of fluorescent-labelled cells which offer a unique insight into inflammatory processes in the eye. The facility to image with several different wavelengths simultaneously offers the potential for spectral imaging of retinal tissue with the aim of revealing those early changes in tissue perfusion that indicate the onset of retinal disease, so increasing the probability of successful therapy.

Animals↗

Improved leukocyte tracking in mouse retinal and choroidal circulation.

The purpose of this study is to develop a new method with which to visualize leukocyte dynamics in murine choroidal and retinal circulation. Both pigmented (B10.RIII) and non-pigmented (BALB/c) mice were used in this study. One hundred microl of 0.05% sodium fluorescein was injected via the mice tail vein to outline the vessel, followed by 150 microl (10(7) cells) C-AM labelled leukocytes. Fundus images were obtained with a confocal scanning laser ophthalmoscope. The dynamic image sequences were recorded simultaneously on videotape (S-VHS) and digitally at 25 frames per sec. The digital images were later analysed with a custom-made personal computer-based image analysis system. Both the choroidal and retinal circulation can be visualized in non-pigmented mice in the first few seconds of fluorescein angiography. However, the view of the choroidal and the retinal capillary circulation is soon blurred due to the rapid fluorescein leakage in the choroid. In contrast, in pigmented mice, retinal circulation is clear against the dark background of the choroid, while choroidal circulation is masked behind the pigment epithelial layer and cannot be seen at all. C-AM labelled leukocytes were clearly seen in the retinal circulation of all experimental mice and in the choroidal circulation of non-pigmented mice for as long as 30 min. The number of labelled circulating cells decreased as time clasped. Cells moved rapidly in the retinal arteries, slowing down or even stopping for a few seconds in the capillary system, and then moved slightly faster again through the postcapillary venules and veins. In non-pigmented mice, significant number of cells were seen to have arrested in the choroidal circulation. There was no difference between B10.RIII mice and BALB/c mice in vessel diameters, leukocyte velocities and shear stresses. This method allows the visualization of leukocytes and provides data on their behavior as they move through the choroidal and retinal circulation of non-pigmented mice, and in the retinal circulation of pigmented mice. It provides a valuable new tool for the investigation of real time leukocyte dynamics in murine retinal and choroidal microcirculations both under physiological conditions and during the development of ocular disease.

Animals↗

True colour imaging of the fundus using a scanning laser ophthalmoscope.

Currently retinal imaging is performed with the fundus camera. This has a number of limitations, in particular the high level of illuminations required for imaging. The scanning laser ophthalmoscope (SLO) has been proposed as an alternative imaging device but to date one of its main limitations has been that it gives only monochromatic images. In this paper we describe an SLO which uses low power red, green and blue lasers to image the human fundus. Using three lasers simultaneously to produce a colour image will increase the fundus exposure by a factor of three. To overcome this problem, a technique has been developed for multiplexing the lasers so that each point on the retina is imaged by the three lasers pulsed rapidly in sequence. The total exposure is thus kept to the same level as for a single laser and total imaging time is not increased. An example is shown of the image from a patient with diabetic retinopathy.

Calibration↗

Clinical investigation of a true color scanning laser ophthalmoscope.

OBJECTIVE: To show true color scanning laser ophthalmoscope (SLO) images produced by simultaneously imaging the retina with red, green, and blue lasers. METHODS: Low-power red, green, and blue lasers were combined using fiber optics. By rapidly pulsing the lasers, each point on the fundus is illuminated by the 3 colors in quick succession, with the total power level being similar to that from a single laser. The reflected light is then decoded to extract the red, green, and blue color information and the true color fundus image is displayed live on a computer monitor. RESULTS: Comparison was made between the color SLO images from 5 patients and their digitized fundus photographs. The background fundus and retinal vasculature showed a similar appearance. The SLO gave better quality information in patients with ocular histoplasmosis, macular dystrophy, and optic disc drusen. By operating the color SLO in the indirect mode, macular edema could be clearly seen as lines and ridges surrounding the fovea. CONCLUSION: The color SLO offers all the advantages of the present commercially available monochromatic device, with the added advantage of true color representation of the fundus without increasing either imaging time or the level of exposure.

Adult↗

Characterization of fine particulate matter produced by combustion of residual fuel oil.

Combustion experiments were carried out on four different residual fuel oils in a 732-kW boiler. PM emission samples were separated aerodynamically by a cyclone into fractions that were nominally less than and greater than 2.5 microns in diameter. However, examination of several of the samples by computer-controlled scanning electron microscopy (CCSEM) revealed that part of the PM2.5 fraction consists of carbonaceous cenospheres and vesicular particles that range up to 10 microns in diameter. X-ray absorption fine structure (XAFS) spectroscopy data were obtained at the S, V, Ni, Fe, Cu, Zn, and As K-edges and at the Pb L-edge. Deconvolution of the X-ray absorption near edge structure (XANES) region of the S spectra established that the dominant molecular forms of S present were sulfate (26-84% of total S) and thiophene (13-39% of total S). Sulfate was greater in the PM2.5 samples than in the PM2.5+ samples. Inorganic sulfides and elemental sulfur were present in lower percentages. The Ni XANES spectra from all of the samples agreed fairly well with that of NiSO4, while most of the V spectra closely resembled that of vanadyl sulfate (VO.SO4.xH2O). The other metals investigated (i.e., Fe, Cu, Zn, and Pb) also were present predominantly as sulfates. Arsenic was present as an arsenate (As+5). X-ray diffraction patterns of the PM2.5 fraction exhibit sharp lines due to sulfate compounds (Zn, V, Ni, Ca, etc.) superimposed on broad peaks due to amorphous carbons. All of the samples contain a significant organic component, with the loss on ignition (LOI) ranging from 64 to 87% for the PM2.5 fraction and from 88 to 97% for the PM2.5+ fraction. Based on 13C nuclear magnetic resonance (NMR) analysis, the carbon is predominantly condensed in graphitic structures. Aliphatic structure was detected in only one of seven samples examined.

Air Pollution↗

Tomographic reconstruction of the retina using a confocal scanning laser ophthalmoscope.

Retinal imaging with a confocal scanning laser ophthalmoscope (cSLO) involves scanning a small laser beam over the retina and constructing an image from the reflected light. By applying the confocal principle, tomographic images can be produced. However, the thickness of such slices, when compared with the retinal thickness, is too large to give useful 3D retinal images. In this study an algorithm has been developed which fits a double Gaussian curve to the axial intensity profiles generated from a stack of image slices. The underlying assumption is that the laser light has mainly been reflected by two structures in the retina, the internal limiting membrane and the retinal pigment epithelium. From the fitted curve, topographic images and novel thickness images of the retina can be generated. The technique has been applied to three normal volunteers and seven patients with macular pathology (cystoid macular oedema and macular hole) demonstrating the clinical value of the technique. The improvement in accuracy achieved by using a double rather than a single Gaussian is also demonstrated.

Humans↗

Quantifying changes in retinal circulation: the generation of parametric images from fluorescein angiograms.

Fluorescein angiography is an established technique for examining the functional integrity of the retinal circulation. The ability to quantify this function offers the possibility of early detection of changes due to retinopathy. We have developed a technique to generate functional, parametric images of the retinal circulation. A given angiogram is first registered to align consecutive frames. At each point in the retina, a graph of fluorescein intensity versus time is then constructed and fitted with a gamma variate curve. Parameters are extracted from these curves and formed into parametric images showing the variation in fluorescein passage across the entire area of the angiogram. Parameters examined to date include time to maximum intensity, time of arrival and rise time. The technique has been demonstrated using photographic and scanning laser ophthalmoscopic angiograms of both normal subjects and patients with a variety of retinopathies. The time to maximum images of the normal subjects reveals a similar fillings pattern in each case, whilst the pathologies present in the abnormal angiograms are clearly identified. The generation of functional time to maximum images enables the health of the retinal circulation to be quantified with respect to the rate at which the vasculature fills with fluorescein. This offers a potential tool for detecting the onset of retinopathy and monitoring its progression.

Fluorescein Angiography↗

Novel approach towards colour imaging using a scanning laser ophthalmoscope.

AIMS: Conventional fundus imaging using a fundus camera produces colour fundus pictures. The scanning laser ophthalmoscope (SLO) has the advantages of lower levels of light exposure, improved contrast, and direct digital imaging but until now has produced monochromatic images as a laser of single wavelength is used. True representation of the fundus is possible by combining images taken using blue, green, and red lasers. METHODS: A custom built SLO was used to capture blue, green, and red fundus images from suitable volunteers and patients with fundus disease. Images were corrected for eye movement and combined to form a colour image. Colour fundus photographs were taken using a fundus camera for comparison with the SLO image. RESULTS: The background fundus and retinal vasculature had similar appearances with the two imaging modalities. Internal limiting membrane reflections were prominent with the SLO. Identification of new vessels in the diabetic fundus was easier with the SLO than the colour fundus photographs. CONCLUSION: A colour SLO offers all the advantages of the present monochromatic imaging system with the added advantage of true colour representation of the fundus.

Adult↗

In vivo cell tracking by scanning laser ophthalmoscopy: quantification of leukocyte kinetics.

PURPOSE: To image peripheral blood leukocyte traffic in the normal retinal and choroidal vasculature and to quantify the differences in the circulation dynamics between normal and concanavalin A (ConA)-activated leukocytes. METHODS: Normal or ConA-activated splenocytes were fluorescently labeled in vitro with 6-carboxyfluorescein diacetate (CFDA) and reinfused in vivo where they were tracked in the retinal and choroidal circulations of syngeneic rats by means of a scanning laser ophthalmoscope (SLO). Simultaneous digital and video images were captured for as long as 30 minutes, and the initial 15 seconds of image sequences and leukocyte dynamics were analyzed from digitized images by recording the velocity of trafficking cells and the number of stationary cells that accumulated with time, using a customized software package. RESULTS: Mean velocity (+/-SD) was 29.8 +/- 15.3 mm/sec in the retinal arteries, 14.7 +/- 7.2 mm/sec in the retinal veins, and 3.0 +/- 3.6 mm/sec in the retinal capillaries. Mean velocity in the choroidal vessels was 6.1 +/- 6.0 mm/sec. No significant difference in leukocyte velocity was found between activated and normal leukocytes in any of the vessel systems. However, activated leukocytes were observed to accumulate more within the choroidal vasculature (P < 0.001) and the retinal capillaries (P < 0.001) than in control animals, but not in larger retinal vessels. CONCLUSIONS: A technique to measure the kinetics of circulating leukocytes in vivo has been developed. Although leukocyte activation itself is insufficient to cause slowing of leukocyte velocity, the data indicate that leukocyte adherence to endothelium can be induced in the absence of local or systemic activating stimuli.

Animals↗

The scanning laser ophthalmoscope.

The imaging of the fundus of the eye poses two major technical challenges. First, it is necessary for both the illuminating and reflected beams to pass through the same aperture, the iris. In some commonly used instruments this leads to the use of levels of illumination close to the maximum tolerable by a patient. Second, in order to visualize the different structures present in the various layers of the fundus it is necessary to perform tomographic imaging. The scanning laser ophthalmoscope provides an answer to these particular problems. By scanning the fundus with a narrow laser beam most of the area of the iris is then available for the reflected light and so the intensity of the illuminating beam can be kept low, making it more acceptable for patients. The use of confocal imaging allows 3D images to be produced. In this short review the performance of the instrument will be discussed and its application to a number of clinical problems in ophthalmology considered. Finally there will be a brief description of other instrumentation currently under development.

Diagnostic Imaging↗

Imaging of optic nerve head drusen with the scanning laser ophthalmoscope.

BACKGROUND: Optic nerve head drusen may present diagnostic difficulties in cases of disc swelling. Imaging of the nerve in a search for drusen is often inconclusive, especially in children, where drusen may be buried below the surface of the nerve head. METHODS: A small study was carried out using a scanning laser ophthalmoscope (SLO) with an infrared confocal facility to scan deep within optic discs in an attempt to image drusen. RESULTS: The SLO was able to demonstrate superficial and buried drusen (using the infrared confocal facility). The superiority of the SLO over ultrasound in the presence of lens opacity was revealed, as the SLO simultaneously demonstrated both drusen and the associated anomalous disc features which are not detected by ultrasound. CONCLUSION: The SLO can help in the diagnosis of optic disc drusen especially in difficult cases where lens opacity or buried drusen hinders their definitive diagnosis.

Adolescent↗

Quantitative image analysis of macular drusen from fundus photographs and scanning laser ophthalmoscope images.

Drusen are common features in the ageing macula and are associated with exudative age-related macular degeneration (ARMD). They appear as characteristic structures in indirect mode infrared scanning laser ophthalmoscope (SLO) images. Manual counting of drusen area in SLO images has shown no significant difference from that in standard fundus photographs in 6 eyes of 5 patients (p > 0.5). Computerised image processing techniques have been applied to digitised colour fundus photographs and indirect mode images to quantify the area of the macula affected by drusen in an automated fashion. Application of these methods may permit objective and repeatable assessment of the natural history of macular drusen. For a specificity of 90%, a sensitivity of 60% for colour fundus photographs and 35% for SLO images has been achieved, when compared with manual counting. The colour fundus photograph method also showed superior reproducibility compared with the SLO technique.

Aged↗

Fundus imaging in patients with cataract: role for a variable wavelength scanning laser ophthalmoscope.

AIMS: An investigation was carried out to compare the image quality of the ocular fundus obtained clinically, photographically, and with the scanning laser ophthalmoscope (SLO) at visible and infrared wavelengths in patients with significant cataract. METHODS: Nineteen patients admitted for routine cataract extraction were examined clinically by two independent observers to ascertain cataract type and clarity of fundus view with an indirect ophthalmoscope. Fundus photography and both confocal and direct (non-confocal) SLO imaging at 590 nm, 670 nm, and 830 nm were carried out after pupillary dilatation. Images obtained were graded independently using a recognised grading system. RESULTS: Quality of SLO images appeared to be superior to indirect ophthalmoscopy (p < 0.01) and fundus photography (p < 0.001) when graded subjectively. Quantitative analysis of contrast of retinal vessels demonstrated significantly higher contrast for the SLO compared with digitised fundus photographs at all wavelengths tested (p < 0.001), with highest contrast at 590 nm. Use of a confocal aperture significantly improved vessel contrast but may reduce overall image intensity. CONCLUSIONS: Scanning laser ophthalmoscopy may offer a method to observe and record fine fundus detail in patients who have marked cataract.

Cataract↗

Performance measurements of an infrared digital scanning laser ophthalmoscope.

Direct digital acquisition of images using a scanning laser ophthalmoscope (SLO) offers several advantages over the conventional fundus camera; in particular, the ability to produce tomographic images using a confocal aperture. This note describes measurements of the performance of an SLO. Spatial resolution, measured by the modulation transfer function, MTF, was shown to be worse along the direction of scan. As expected, image uniformity was good, with a coefficient of variation of 1.9%. While the effect of using a 100 microns diameter confocal aperture instead of one with a 400 microns diameter was to reduce slice thickness from 2600 microns to 975 microns, image intensity was reduced by a factor of 30.

Humans↗

Clinical investigation of an infrared digital scanning laser ophthalmoscope.

An infrared scanning laser ophthalmoscope (SLO) has been custom built in order to investigate the application of confocal and indirect mode SLO imaging to patients with fundus disease. Infrared light is reflected from the fundus to a greater extent than visible light permitting lower illumination power and, as it penetrates the retinal pigment epithelium, choroidal structures can be readily imaged. Furthermore, as conventional infrared illumination and detection systems are not well suited to ophthalmoscopy, this area is underdeveloped as a potential source of useful clinical data. Confocal, direct and indirect imaging modes have been used to image fundi of normal volunteers and patients with fundus disease. In comparison with conventional fundus photography confocal infrared SLO imaging improves visualisation of choroidal vasculature, retinal pigment epithelial abnormalities, laser photocoagulation scars, and optic disc pores in the lamina cribrosa. Direct infrared SLO imaging enables fundus visualisation through nuclear lens opacities. Furthermore, indirect mode imaging enhances significantly the appearance of macular drusen. The potential clinical benefit of these observations is discussed.

Adult↗

Digital fundus imaging using a scanning laser ophthalmoscope.

While many systemic diseases have ophthalmic manifestations with visible pathological features affecting the fundus, diabetic eye disease is of particular interest. There are many advantages in using digital image analysis to quantify the extent of retinal pathology in diabetes particularly to monitor its progression. However, the usual method of imaging involves photographing the retina using a fundus camera. Not only does this require that the photographs be digitized prior to analysis, but the resulting illumination across the image is non-uniform and this creates problems in digital image analysis. This paper describes the construction of a scanning laser ophthalmoscope interfaced to a computer, which has a number of significant advantages for digital retinal imaging. Firstly, it has better uniformity of illumination than the fundus camera and operates at much lower light levels. Secondly, it permits imaging at various wavelengths with no consequent degradation in image quality; in particular, the value of imaging at infrared wavelengths to give greater tissue penetration is demonstrated. Finally, by introducing the capability for confocal imaging, tomographic images can be taken, improving the perceptibility of features situated deep in the retina. Typical images are shown to demonstrate the potential of this instrument.

Diabetic Retinopathy↗

A double-blind crossover trial of atenolol, enalapril and the fixed combination of atenolol and nifedipine in mild and moderate hypertension.

Sixty-three patients with mild to moderate hypertension completed a double-blind, randomised crossover trial of atenolol 100 mg daily, enalapril 20 mg daily and the fixed combination atenolol 50 mg and nifedipine retard 20 mg (Tenif) once daily. In the three areas of efficacy that the study compared-24-hour post dose blood pressure, control of blood pressure over the dosing period with particular reference to the waking day and attenuation of systolic blood pressure on exercise-atenolol and the fixed combination demonstrated greater efficacy than enalapril.

Atenolol↗