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PubMed1991

Helicobacter pylori in gastric biopsy specimens. Comparison of culture, modified giemsa stain, and immunohistochemistry. A retrospective study.

Antral biopsy specimens of 302 different endoscopic investigations of 200 patients with non-ulcer dyspepsia were studied for the presence of Helicobacter pylori in order to determine the most sensitive detection method. Part of the biopsy was cultured, and part stained using a modification of the Giemsa stain, and with an immunoperoxidase technique using a polyclonal rabbit anti-H. pylori antiserum. Cross-reactivity of this antiserum with other Campylobacter species was minimal. Material from 244 investigations was studied using all three detection methods. Culture was positive in 44 per cent, Giemsa in 78 per cent, and immunoperoxidase in 89 per cent of these biopsy specimens. Only five positive Giemsa stains with negative immunoperoxidase stain were found, whereas in 32 cases a negative Giemsa stain with a positive immunoperoxidase stain was seen. In the latter cases, the bacterial load was very low. The specimens revealed bacteria only sporadically, always confined to the deep layers of the gastric pits. Culture results correlated significantly with the bacterial load observed in the Giemsa stain. It is concluded that culture of H. pylori is the least sensitive detection method, whereas immunoperoxidase staining is the most sensitive. For daily practice the modified Giemsa stain, however, appears to be sufficient to diagnose the presence of the micro-organism.

Adolescent↗

PubMed1988

[Expert opinion on biologic stains. Determination of status, future trends].

In this paper an attempt is made to critically review the literature, with special emphasis on bloodstain analysis. One essential aim is the integration of this field into casework. Three basic components in skillful assessment of stains are described: (1) analysis of stain morphology, (2) discriminating and attributing analyses, (3) individualization. Regarding the first, the analysis of stain morphology is based upon the extensive experimental literature published since 1895--mainly in continental Europe. Since 1971 there have also been publications in the American literature. The large family of stain forms and their dependency on multiple variables are described, especially regarding the modes of formation, the energy of impact, and the physical properties of the substrate. The essential elements for reconstruction of the crime are described. The areas of application are arranged in case groups. Since in case work the stain pattern is complicated by many artifacts and overlaps, forensic pathologists are considered the ideal experts for the analysis of bloodstain patterns, as they have a profound knowledge of the type and sequence of injuries. If this is not the case, the forensic pathologist should at least be integrated into the investigating team. In practical application, the stain form is not always adequately analysed. The education and training of pathologists should be improved to achieve this standard. Analysis of the stain morphology and a subsequent selection of stains are also essential prerequisites for meaningful further investigations. By the use of discriminating and attributing analyses, one can as a rule arrive at a definite answer by using only one test. This is true for basic questions such as the identification of blood type, as well as proof of exclusion. One can distinguish between traditional methods, the new field of immunochemistry and rarely used methods. Immunochemistry has permitted success in recent years in determination of the blood group from hair. It is recommended that reference laboratories be established for training in these rare methods. Individualization analyses are subdivided into two large fields: non-DNA individualization and DNA individualization. It is postulated that in the future stain laboratory both areas will coexist. In non-DNA individualization, essential progress has been made. The detection of protein polymorphisms by blotting and subsequent visualization by antibody-linked enzyme/substrate reactions has led to a considerable increase in sensitivity and specificity.(ABSTRACT TRUNCATED AT 400 WORDS)

Blood Stains↗

PubMed1977

Differential staining of polytene chromosome bands in Chironomus by Giemsa banding methods.

Two Giemsa banding methods (C banding and RB banding) are described which selectively stain the centromere bands of polytene salivary gland chromosomes in a number of Chironomus species. - By the C banding method the polytene chromosome appearance is changed grossly. Chromosome bands, as far as they are identifiable, are stained pale with the exception of the centromere bands and in some cases telomeres, which then are intensely stained reddish blue. - By the RB method the centromere bands are stained bright blue, whereas the remainder of the polytene bands stain red to red-violet. - Contrary to all other species examined, in Chironomus th. thummi numerous interstitial polytene chromosome bands, in addition to the centromere regions, are positively C banded and blue stained by RB banding. In the hybrid of Ch. th. thummi X Ch. th. piger only those interstitial thummi bands which are known to have a greater DNA content than their homologous piger bands are C banding positive and blue stained by the RB method whereas the homologous piger bands are C banding negative and red stained by RB banding. Ch. thummi and piger bands with an equal amount of DNA both show no C banding and stain red by RB banding. - It seems that the Giemsa banding methods used are capable of demonstrating, in addition to centromeric heterochromatin, heterochromatin in those interstitial polytene chromosome bands whose DNA content has been increased during chromosome evolution.

Animals↗

PubMed1989

Immunocytochemical detection of sulphonylated DNA in tissue sections. An alternative to Feulgen staining of DNA.

We report here on a new sensitive and highly specific DNA staining technique which we have called sulpho-DNA staining. DNA staining is based on a sulphonylation reaction of 2'-deoxycytidine or cytidine that takes place in the 6th position of cytosine with ensuing immunodetection of the sulphonylated DNA. The specificity of DNA staining is introduced by the use of an antibody recognizing only modified DNA but not modified RNA, by recourse to an additional acid hydrolysis step which destroys RNA but not DNA. We describe here the optimal conditions for the sulphonylation of DNA using O-methylhydroxylamine and metabisulphite as reactants. The new DNA stain labels all nuclei in either normal human tissue or in tumor cells. For nuclear DNA the staining signal is higher for the sulpho-DNA staining than for the Feulgen staining for nuclear DNA. This new DNA staining technique is suitable for use on tissue sections as well as on cytosmears.

Animals↗

PubMed1979

Evaluation of the glyoxal-bis-(2-hydroxyanil)-method for staining of calcium in model gelatin films and pancreatic islets.

The nature of tissue calcium, detectable with glyoxal-bis-(2-hydroxyanil), (GBHA), was investigated using gelatin films as model. The results indicate that in the films the procedure detects only the calcium fraction which was ionized in the original gelatin solution. The GBHA staining intensity (absorbance) appeared to be linear with the amount of ionized calcium in the range from 0 to 2 micrograms/cm2. The method allows detection of amounts of ionized calcium as low as 0.15 micrograms/cm2 or 0.0015 pg/mu2. For the measurement of calcium in pancreatic tissue of fed rats, the tissue was subjected to freeze-substitution at -80 degree C in acetone containing 1% oxalic acid. Adjacent sections were stained with either GBHA or aldehyde-fuchsin (AF). Exocrine tissue hardly stained with GBHA whereas islet tissue stained intensely. For GBHA as well as for AF a variation in staining intensity (visual evaluation) between islets was observed. Islet GBHA- and AF-staining intensities did not correlate. The AF-staining intensity but not the GBHA-staining intensity decreased with increasing islet diameter. Also in pancreatic islet tissue the GBHA method appears to be very sensitive and reproducible and small differences in islet GBHA-staining intensity can be detected. The results indicate that between islets differences in ionized calcium content exist. These differences do not correlate with the degree of B-cell granulation.

Animals↗

PubMedDate not supplied

Evaluation of housestaff physicians' preparation and interpretation of sputum Gram stains for community-acquired pneumonia.

OBJECTIVE: To evaluate the preparation and interpretation of sputum Gram stains by housestaff physicians in the assessment of patients with community-acquired pneumonia. DESIGN: A prospective, multicenter study. SETTING: Two university-affiliated hospitals in Pittsburgh. PATIENTS: Ninety-nine cases of clinically and radiographically established pneumonia occurring in 97 patients. Diagnostic test assessment: Housestaff and microbiology personnel prepared a Gram stain for each case of pneumonia. Housestaff assessed the presence and identity of a predominant microbial organism on the slides they prepared. Two senior staff microbiologists, blinded to patient and preparer, evaluated all slides for preparation, sputum purulence, and identification of the predominant organism. Two reference standards were used to assess the sensitivity, specificity, and predictive values of housestaff's Gram-stain interpretations: 1) senior staff microbiologists' determinations of the microbes present using the slides without benefit of culture results, and 2) the etiologic agent derived from results of sputum culture, blood culture, or serology. MEASUREMENTS AND MAIN RESULTS: Housestaff physicians completed a Gram stain in 58% of the pneumonia episodes. Gram stains were not made in 42% of cases, primarily because patients were unable to produce sputum. Fifteen percent of housestaff's smears were judged inadequately prepared, compared with 3% for the laboratory personnel (p less than 0.01). Housestaff obtained purulent sputum samples significantly more often than did nursing personnel (58% versus 38%; p less than 0.01). Housestaff's Gram stains were 90% sensitive for detecting pneumococcus, with a 50% false-positive rate. The sensitivity of the Gram stain was less for identification of Haemophilus influenzae than for identification of Streptococcus pneumoniae. A single antimicrobial agent was chosen as initial therapy for 50% of the patients in whom housestaff identified a predominant organism, compared with 30% in whom a predominant organism was not identified (p less than or equal to 0.05). CONCLUSIONS: Although housestaff obtained purulent sputum samples more frequently than did nursing personnel, they made systematic errors in the preparation and interpretation of Gram-stained slides. Housestaff physicians should receive formal training in the preparation and interpretation of Gram stains; the specific defects elucidated in this study warrant special attention.

Adolescent↗

PubMed2003

Long-term staining of live Merkel cells with FM dyes.

Live Merkel cells in the skin and hair follicles are known to incorporate a fluorescence dye, quinacrine, which has been utilized to identify and dissect the cells for experiments. Quinacrine fluorescence of the cells is, however, quickly lost and quinacrine-stained Merkel cells soon become difficult to identify in tissue culture. To find dyes that remain in the cells for a long period of time, we tested many fluorescence dyes and found that FM dyes (such as FM1-43) are useful markers for live Merkel cells. In the rat footpad skin, FM1-43 was shown to stain 95% of live Merkel cells that were already stained with quinacrine. FM4-64 stained 98% of quinacrine-stained Merkel cells. Merkel cells in sinus hair follicles were also stained with FM dyes. The fluorescence intensity of FM dyes was stronger than that of quinacrine, and the shape of the cells was more distinct in the FM-dye-stained cells. To test how long FM dyes remain in live cells, FM-dye-stained Merkel cells in hair follicles were embedded in collagen gel and were cultured in a serum-free medium. FM-dye-stained cells were easily identified even after 7 days of culture. During the culture, Merkel cells changed their shape, moved in the preparation and tended to aggregate on the surface. We conclude that FM dyes are powerful tools for tracing live Merkel cells in in vitro experiments. Moreover, the finding that Merkel cells incorporate FM dyes suggests that vesicles in the cells are likely to have mechanisms of recycling in a manner similar to those in neurons and secretory cells.

Animals↗

PubMed1985

Sialoglycoproteins with a high amount of O-glycosidically linked carbohydrate moieties stain yellow with silver in sodium dodecyl sulfate-polyacrylamide gels.

Several sialoglycoproteins and human salivary proteins were analyzed in sodium dodecyl sulfate-polyacrylamide gels using the silver/Coomassie-staining protocol (J. K. Dzandu, M. E. Deh, D. L. Barratt, and G. E. Wise, 1984, Proc. Natl. Acad. Sci. USA 81, 1733-1737) to determine the extent to which yellow Ag staining originally reported for human red blood cell glycophorins can be applied to other sialoglycoproteins. Results showed that not all sialoglycoproteins elicit a positive yellow color in the silver stain reaction. Some of the sialoglycoproteins stained as brown or negative images in the Ag-staining cycle. Alkaline beta elimination of O-glycosidically linked carbohydrate chains of glycophorin resulted in the loss of yellow color development in the Ag-staining protocol. Analysis of acidic salivary proteins showed several yellow Ag-stained bands at Mr X 10(-3) = 150, 82, 70, 51, 46, and 42. These results suggest that the carbohydrate moieties of glycophorin removable by alkaline beta elimination are responsible for the characteristic yellow color in the Ag stain reaction. In addition, under our staining conditions sialoglycoproteins with a high amount of O-glycosidically linked carbohydrate chains give a characteristic yellow silver stain.

Carbohydrates↗

PubMed1990

Staining of free sulfhydryl groups of proteins after separation by isoelectric focusing of Bio-Rad standards and lens crystallins.

In this study, we propose a new staining method for free sulfhydryl groups of proteins after having separated native samples by thin-layer isoelectric focusing (IEF) in absence of detergents. A comparison was made between proteins stained purple for free sulfhydryl groups (SH) and proteins stained blue by Coomassie blue (CB). A stainability factor, F = %SH/%CB, was calculated for each protein. The Bio-Rad IEF standards containing seven marker proteins for pH scale determination were stained purple, in the same way as they were designed for CB staining. To prove the validity of the currently proposed staining method for a defined protein system such as the eye lens crystallins, these proteins were also stained after IEF as described above. The factor F was calculated for all alpha-, beta-, and gamma-crystallin components that stained in both methods. We discovered that alpha-crystallins contained comparatively high amounts of free SH groups, while some beta- and gamma-crystallin components also contained considerable amounts of free SH groups. The SH staining procedure with 2,2'-dihydroxy-6,6'-dinaphthyl disulfide applied after IEF appeared to be useful, specific, and reproducible.

Animals↗

PubMed1978

A quantitative electron microscopic study of synapse formation in dispersed cell cultures of rat cerebellum stained either by Os-UL or by E-PTA.

Synapse formation was followed in dispersed cell cultures of rat cerebellums stained either by osmium-uranyl-lead (Os-UL) or by ethanolic phosphotungstic acid (E-PTA). The numerical densities of synapses stained by Os-UL were always significantly higher than those stained by E-PTA (from 3 to 35 days in vitro). This difference suggests that some portion of the populations of both immature and mature synapses was not stained by E-PTA. The width of the synaptic cleft (28.4nm) in synapses stained by E-PTA was only 9nm more than that of the cleft in synapses stained by Os-UL (19.4nm), suggesting that some portion of one or both of the synaptic membranes is stained by E-PTA. Analysis of data from 7 other staining procedures utilizing both ethanolic and aqueous solutions demonstrated that the differences in cleft width described above appear to be due to the various affinities of the stains for different portions of synaptic membranes, and do not represent differences due to shrinkage artifact. In examining the parameters of synaptic structure during development of the cultures, a statistically significant increase in both the height and width of the presynaptic dense projections was found. Changes in synaptic morphology during synaptogenesis in this culture system were similar to those reported for the cerebellar cortex in vivo.

Animals↗

PubMed1979

E-PTA stains oligodendroglial surface membranes and microtubules in optic nerves during myelination.

Aldehyde fixed Xenopus tadpole and frog optic nerves were stained en bloc with ethanolic phosphotungstic acid (E-PTA). During rapid myelination, intense staining was observed on cytoplasmic faces of paranodal terminal loops and loosely wrapped oligodendroglial membranes found along inner and outer surfaces of compact myelin sheaths. Oligodendroglial microtubules also were heavily stained. Where stained cytoplasmic faces fused to form a lamella of compact myelin, the intense staining was reduced to a thinner, fainter line. In optic nerves of adult frogs, the staining was less dense but the pattern was similar. The staining distribution and available histochemical evidence indicate that E-PTA stains positively charged proteins non specifically. Since myelin basic protein is found in oligodendroglia during myelination, we suggest that it is being stained by E-PTA while being transported along microtubules to sites where it is inserted into developing myelin lamellae.

Animals↗

PubMed1979

Age-dependent variability of ribosomal RNA-gene activity in man as determined from frequencies of silver staining nucleolus organizing regions on metaphase chromosomes of lymphocytes and fibroblasts.

Frequencies of silver staining nucleolus organizing regions (NORs) have been determined in lymphocytes and fibroblasts from very young and from aged persons. Since silver staining of NORs is associated with activity of ribosomal RNA-genes, we used this approach to investigate a possible inactivation of these genes during aging. Our lymphocyte data are based on a study per age-group of 220 metaphases from 10 subjects. Although in both age-groups modal numbers of silver staining chromosomes per metaphase had similar ranges over the subjects, the frequency of metaphases containing the maximal number of staining chromosomes was in the old age-group (80--89 years) significantly lower than in the young age-group (less than 1 year old). In fibroblasts, of which 75 metaphases from 4 subjects were included per age-group, differences were more pronounced. Modal numbers of silver staining chromosomes were for the aged persons (69--83 years) lower than for the young children (less than 1 year old). Highly significant differences were observed between both groups in frequency of metaphases containing the maximal number of positively reacting acrocentric chromosomes and, more in general, in frequencies of silver staining D- and G-group chromosomes, the lower frequencies being found in the old age-group. We propose the term NOR-junctions as distinct from satellite associations for arrangements of acrocentric chromosomes which after silver staining are visibly connected at their NORs. The number of acrocentric chromosomes involved in lymphocyte NOR-junctions of aged people was significantly higher than the number of joined acrocentrics in young children. The frequency of these NOR-junctions themselves, irrespective of the number of chromsomes involved, was higher for aged persons than for young children, although this difference appeared to be statistically not significantly higher than in fibroblasts. Also based on qualitative observations from our study we discuss tcehnical and biological problems of our approach to study cell aging in vivo by means of silver staining of NORs. We conclude that in man, reflected by the difference in frequencies of silver staining NORs between young and aged persons, a rather extensive loss of ribosomal RNA-gene activity may occur during aging.

Aged↗

PubMedDate not supplied

Intra-axonal Neurobiotin injection rapidly stains the long-range projections of identified trigeminal primary afferents in vivo: comparisons with HRP and PHA-L.

Currently available methods for studying the morphology of physiologically characterized primary afferents are limited by difficulties inherent in impaling thin fibers and by the limited distances over which conventional tracers move during the course of a recording session. We have encountered an alternative method that overcomes these limitations. Neurobiotin (NB; Vector) injections into rat trigeminal (V) primary afferents in the brain stem or V ganglion provided rapid, long-range staining with recording and electrophoretic parameters that are commonly used to eject horseradish peroxidase (HRP) or Phaseolus vulgaris leucoagglutinin (PHA-L). When NB was injected into brain stem fibers responsive to vibrissal deflection with A-beta conduction velocities, collaterals were darkly stained in each of the 4 V subnuclei, as well as the cervical dorsal horn. Labeled fibers were also seen in the V root and peripherally in the infra-orbital nerve for a distance up to 15 mm from the injection site (30 mm total). Cell bodies in the ganglion were never labeled. When NB was injected into V ganglion cells with low- or high-threshold receptive fields and A-beta or A-delta conduction velocities, parent axons were stained in the V spinal tract to the level of the obex, and collaterals were visible in each of the 4 V subnuclei. Such long-range staining occurred within 4 h of tracer injection. HRP never stained brain stem fibers following ganglion cell injections and, when injected centrally with the same survival intervals used with NB, dark staining was limited to within 4 mm of the injection site. Unlike NB or HRP, PHA-L injections rarely produced useful data, either because of the high mortality accompanying attempts to achieve a 1-2 week survival period or because injected neurons were not recovered. Due to its rapid and robust transport, NB is a more convenient and reliable tracer than PHA-L for producing long-range staining of the projections of identified ganglion cells. Intracellular injection of NB also produces rapid Golgi-like staining of fibers over much greater distances than HRP under equivalent staining parameters.

Afferent Pathways↗

PubMed1992

A new electron microscope positive staining method for viruses in suspension.

A new procedure for the positive staining of viruses in suspension, the Tokuyasu staining procedure (TSP), was evaluated using a non-enveloped virus, rotavirus; an enveloped virus, rubella virus and two glutaraldehyde-treated enveloped viruses, Human T Cell Lymphotropic Virus Type I (HTLV-I) and Human Immunodeficiency Virus Type 1 (HIV-1) as models. The TSP involves an initial staining of the virus with uranyl acetate (UA) followed by thin embedding in a mixture of UA and polyvinyl alcohol (PVA). Using aqueous UA for the TSP, a combination of positively and negatively stained particles was seen for both rotavirus and rubella virus. With glutaraldehyde-fixed HTLV-I and HIV-1, stain penetration did not occur and only negative staining was observed. The substitution of methanolic UA for aqueous UA in the TSP resulted in only positive staining of rotavirus and rubella virus. The change in procedure also resulted in stain penetration of the glutaraldehyde-fixed HTLV-I and HIV-1 to give positively stained particles. Some novel morphological features of rotavirus and rubella virus structure were observed by the TSP.

Glutaral↗

PubMed2006

Clinical diagnosis of cutaneous leishmaniasis: a comparison study between standardized graded direct microscopy and ITS1-PCR of Giemsa-stained smears.

Parasitological diagnosis of cutaneous leishmaniasis is absolutely necessary before treatment. Direct microscopy of scrapings taken from the margins of skin lesions is the most commonly used method for clinical diagnosis of leishmaniasis. In this study to evaluate the usage of stained smears as samples for PCR and the possible advantage of PCR, we compared the sensitivity of the diagnosis of Giemsa-stained skin scrapings by standardized graded direct microscopy with that of ITS1-PCR with the material of the same area of the slide. Three 5mm x 5mm squares were marked on each of the 20 Giemsa-stained touch smears from 20 clinically diagnosed Palestinian patients. Out of the 60 squares scanned for amastigotes under 100x oil-immersion light microscopy, 45 (75%) gave usable results and 23 of these were positive for Leishmania. Fifteen (25%) squares could not be scanned microscopically, 12 because of staining that was too thick and 3 because of inadequate staining. DNA from each scanned square was extracted separately after microscopy and run through ITS1-PCR. Of the 23 microscopy-positive squares, 20 (87%) of these were positive by PCR. Of the three that were negative, one failed to extract for DNA, the second showed only one amastigote in the entire square, and the third was normally graded as +1 but was not amplified for unknown reasons. Of the 22 squares negative for microscopy, 18 (82%) were ITS1-PCR positive. Additionally, all three improperly stained squares were ITS1-PCR positive. Of the 12 darkly stained squares, 11 were positive. A negative control group of 15 German individuals from which Giemsa-stained slides containing three squares each was prepared and these slides were also microscopically scanned and tested by ITS1-PCR. Both tests were negative with both methods. Compared to microscopy (data in parenthesis), PCR showed a sensitivity of 87% (37%) and a specificity of 100% (100%). We have concluded that Giemsa-stained smears are a readily usable sampling method for PCR and that ITS1-PCR is far more sensitive than microscopy.

Adolescent↗

PubMed2006

Comparison of Gram and Kopeloff stains in the diagnosis of bacterial vaginosis in pregnancy.

Bacterial vaginosis (BV) is commonly diagnosed by using the Nugent score, a semiquantitative scoring system to evaluate bacterial morphotypes on Gram stain of vaginal secretions. Some authors have suggested using the Kopeloff modification of the Gram stain. Asymptomatic BV in pregnancy has been associated with adverse outcomes. We performed both stains on simultaneously collected vaginal smears from 2652 women at 24-26 weeks of gestation. Gram staining gave significantly higher (more abnormal) Nugent scores than Kopeloff staining. Compared to the Kopeloff stain, the number of specimens graded as indeterminate or consistent with BV by Gram stain increased by 29% (469 versus 364, P<.001). Interrater reliability of the Nugent score (n=413) for Kopeloff staining was significantly better than Gram staining (agreement=74% versus 63%, intraclass correlation coefficient=0.87 versus 0.79, P<.05, 95% confidence intervals 0.85-0.89 and 0.75-0.82, respectively).

Adult↗

PubMed2004

Effect of blue staining of expandable hydrophilic intraocular lenses on contrast sensitivity and glare vision.

PURPOSE: To evaluate the effect of trypan blue staining of expandable hydrophilic acrylic intraocular lenses (IOLs) on contrast sensitivity and glare. SETTING: Department of Ophthalmology, Federal University, and Oculistas Associados, Rio de Janeiro, Brazil. METHODS: Phacoemulsification with expandable hydrophilic IOL implantation was performed in 19 eyes. Group 1 (stained group) consisted of 12 eyes with the IOL unintentionally stained by trypan blue 0.1% solution during surgery; Group 2 (unstained group) consisted of 7 eyes with IOLs that were not stained because trypan blue was not used during surgery. The eyes were examined postoperatively for Snellen best corrected visual acuity (BCVA), contrast sensitivity BCVA, and glare vision. The examiners were masked to whether the IOL was stained by trypan blue. Inclusion criteria included a BCVA of 20/30 or better 1 month postoperatively. Patients who had other ocular pathology or previous ocular surgery were excluded from the study. RESULTS: The mean postoperative BCVA was 20/24 in the stained group and 20/23 in the unstained group (P =.73). The mean contrast sensitivity visual acuity was 20/31 in the stained group and 20/26 in unstained group (P =.10). The mean glare visual acuity was 20/75 and 20/33, respectively (P =.03). CONCLUSIONS: Patients with expandable hydrophilic acrylic IOLs stained with trypan blue performed significantly worse on a glare vision test than patients whose IOLs were not stained. Cataract surgeons should avoid using trypan blue with this IOL type.

Acrylic Resins↗

PubMed2004

Time course of lens capsule staining using trypan blue and indocyanine green: in vitro study in porcine eyes.

PURPOSE: To determine whether staining of the lens capsule with trypan blue 0.1% and indocyanine green (ICG) 0.5% diminishes with time and whether it differs between the anterior and posterior capsules. SETTING: Department of Ophthalmology, Keio University School of Medicine, Tokyo, Japan. METHODS: Crystalline lenses removed from porcine eyes were stained for 10 seconds with 0.1 mL of trypan blue 0.1% or indocyanine green 0.5%. They were then placed in distilled water and observed for the persistence of staining over time. In a second experiment, the anterior chamber and internal aspects of the anterior capsule and internal and vitreous aspects of the posterior capsule were gently irrigated with 0.1 mL of trypan blue 0.1% or ICG 0.5%. After 10 seconds, the capsules were irrigated with distilled water and the staining intensities were compared. RESULTS: Staining was not diminished 30 seconds, 5 minutes, or 1 hour after application of either dye. No difference was evident in staining intensity or diminution with time between the anterior and posterior capsules, but the external aspects were stained more than the internal aspects with both dyes. Trypan blue produced more intense staining than ICG. CONCLUSIONS: Since the intensity of capsule staining in the intact lens did not change during a 1-hour immersion in water, capsule dyes may not dissipate fully during cataract surgery. As possible long-term adverse effects have not been ruled out, capsule dyes should be used in a low concentration for a short exposure time.

Animals↗