|Year : 2001 | Volume
| Issue : 4 | Page : 273-6
Gross photography of ophthalmic pathology specimens.
J Biswas, K Babu, S Krishnakumar, K Vanitha, P Vaijayanthi
Medical and Vision Research Foundation, Chennai, India
Medical and Vision Research Foundation, Chennai
Source of Support: None, Conflict of Interest: None
Removal of the entire eyeball or a portion of it is required in many ophthalmic conditions. The gross features of such specimens provide valuable information. Gross photography of an intraocular tumour, ruptured eyeball or other pathological specimens including lid lesions, orbital tumors, intraocular lens implants, intraocular and extraocular parasites, retinal pathology such as haemorrhages and peripheral degenerations, establishes a permanent documentation. We herein describe the technique of gross photography of such valuable specimens, using a dissecting microscope with a special arrangement. This technique is currently used by us with gratifying results.
Keywords: Computers, Eye, pathology, Humans, Photography,
|How to cite this article:|
Biswas J, Babu K, Krishnakumar S, Vanitha K, Vaijayanthi P. Gross photography of ophthalmic pathology specimens. Indian J Ophthalmol 2001;49:273
|How to cite this URL:|
Biswas J, Babu K, Krishnakumar S, Vanitha K, Vaijayanthi P. Gross photography of ophthalmic pathology specimens. Indian J Ophthalmol [serial online] 2001 [cited 2020 Mar 29];49:273. Available from: http://www.ijo.in/text.asp?2001/49/4/273/14686
The study of ophthalmic pathology is essential for understanding the basis of eye diseases. The gross features of an excised eye or part of the eye provide valuable information. Permanent documentation of gross features is also necessary for scientific records, communication, teaching and publication.
This article discusses techniques that can be used to make good reproducible gross photographs of ophthalmic pathological specimens.
| Materials|| |
Specimens that can be photographed include a normal eyeball, eyeball with axial abnormalities, tumours and extraocular extensions trauma related changes in the eye retinal lesions (haemorrhages, peripheral degenerations) corneal buttons obtained during penetrating keratoplasty, explanted IOLs, extraocular structures such as excised lid mass, buckle material and intra and extraocular parasites.
The photographic set up includes a 35mm Nikon camera (F3 or FM2 Model, Nikon Corporation, Tokyo, Japan) with a 55mm macro lens (Nikon Corporation, Tokyo, Japan), a Leica Wild M3C dissecting microscope (Leica-Heerbrugg-AGH-9435, Heerbrugg, Switzerland) using cold shadow free illumination system, two armed goose-neck light guides for illumination, a circular jar about 15cm in diameter and 15cm in height and a small cup-shaped stand made to hold the cut sections of the globe [Figure:1]. We use Kodak ektachrome 100 ASA flim for color photographs and Kodak black and white ASA film 100 for black and white photographs.
| Technique|| |
A. Preparation of specimens for gross photography
The formalin-preserved specimen is washed in running water to remove dirt, blood and the strong odour of formalin. It is then transferred to 70-80% ethyl alcohol to restore its colour. One may trim or excise a portion of the adhering structures to clearly expose the area of interest. Clinical description and other investigations, if available, should be looked into. Transillumination of the globe indicates the position and extent of the tumour by a transillumination defect in the globe. Transillumination is done prior to dissection. The globe is held directly in front of bright light, usually a dissecting microscope illuminator or a pencil light. The amount of light transmitted by a normal eye depends on the degree of uveal pigmentation. Transmission is helpful, particularly when the specimen contains intraocular tumors. The eye is manually rotated on the surface of the light until the tumour shadow is most apparent. The shadow is then outlined with a colour marker pen. The location and dimensions of the shadow are recorded. Transillumination is essential to cut open the globe in the proper axis to yield the best view of the tumour. This can be used as a guideline to cut open the eyeball. A laboratory technique devised by Folberg et al can also be followed for a good view of the tumor. The globe is opened adequately to afford the best view of the lesion or the mass.
B. Photographic technique
After performing the routine gross examination with the dissecting microscope and movable illuminator, the specimen is photographed by mounting the body of the camera on the drawtube of the dissecting microscope. The specimen is totally immersed in a petridish containing 50% ethanol (to reduce light reflections and shimmering reflexes). This can be fixed to the lower end of the drawtube of the microscope. The light is then focused on the specimen and the photograph taken at the desired magnification. By using the indirect illumination technique, light is focused to its smallest diameter and directed on the area of interest; subtle features of the specimen can be demonstrated and photographed.
One may use to advantage the auto exposure feature; this calculates the amount of exposure depending on the sensitivity of the film (ASA), and brightness of light. Various types of filters (80 B, blue filter, ND2 grey filter) also can be used. For instance, 80B blue filter is a daylight filter that should be used when daylight films are used. Magnification can also vary - 64, 100, 160, 250, 400 - in the Leica Wild M 3C dissection microscope. However, one loses some amount of resolution when the photograph is taken in higher magnification. The photograph quality can be improved by repeatedly cleaning the specimens with clean water. It is important to keep the microscope objectives and eye pieces clean.
When small lesions in the globe have to be photographed, a photo slitlamp camera can also be used. The specimen is held in front of the lens of the fundus camera with a pair of forceps gripping the back of the specimen and the area of interest is photographed.
An alternative method for gross photography is to use the wooden platform. A colour paper is fixed on the platform and colour of the paper contrasts sharply with that of the specimen. A clear glass plate is placed over the wooden platform with a suitable support. A stand with an adjustable screw clamp, holds the Nikon camera with 55 mm macro lens with 80B filter. The camera is then focused on the specimen on the glass plate. A scale is placed along the length of the specimen to convey dimensions. This is helpful for large specimens with 2 × 250 W photo floodlights.
One can also photograph a stained glass slide with the dissecting microscope by placing it on a white paper for contrast. Otherwise the slide may be placed on an X-ray view box using the white illuminated background and photograph using a standard camera with 55 mm macro lens without filter. Fluorescent light tube balances for daylight in the x-ray view box. Various examples of gross photographs taken using the above techniques are illustrated in [Figure:2] to [Figure:8].
| Digital Photography|| |
While conventional photographic techniques can be quite useful for documentation, the digital facility considerably enhances teaching and publication. Currently used extensively in clinical and basic research, this allows one to attach a digital camera to the dissecting microscope. With the digital camera, the photograph can be viewed immediately to assess its quality. This can also be directly stored in computer. The photograph can be edited using software such as Adobe Photoshop (Abode Systems, Mountain View, California, USA). It can then be cropped, enlarged and edited. Montague and co-workers have used a digital camera technique designed for photomicrography. They placed a glass histopathologic slide in a carrier which was then inserted into 35 mm slide scanner. The slide was scanned to create a digital colour image. Such a colour image can be converted into black & white with enhancement of certain histologic features. This process can provide both 35 mm colour slides or high resolution colour or black and white printouts. The digital camera also allows us to sharpen an out-of-focus image or to convert a positive to a negative image. Many peer-reviewed international journals now accept digital photographs.
However, digital photography is not without disadvantages. For instances, the investigator can edit the photograph to his benefit, and can show the tumour extending or not extending out of the globe, to support a particular thesis. One can also change the colour or shade. The digital camera is more expensive than the standard photography camera.
In summary, the simple procedures described here can be adopted by any ophthalmologist or pathologist. A good gross photograph can add strength to a scientific presentation or communication and can at the same time provide a permanent record.
| Acknowledgement|| |
The authors wishes to acknowledge the contribution of Mr. S.P. Govindarajan, Department of Photography, Sankara Nethralaya, Chennai, for technical assistance.
| References|| |
Eagle R.C. Pathology. YearBook of Ophthalmology
St. Louis: Mosby, 1997;11:341-354.
Folberg.R, Verdick Randy. The gross examination of eyes removed for choroidal and ciliary body melanomas. Ophthalmology
Montague PR, Meyer M, Folberg R. Technique for the digital imaging of histopathologic preparation of eyes for research and publication. Ophthalmology