Capsule Staining Research Articles

Histopathologically proven siderotic cataract with disintegrated intralenticular foreign body

Cataract formation may be an indicator of early siderosis and has been associated with intralenticular foreign bodies. We report a unique case of histopathologically proven lens siderosis in a young man with a preceding history of trauma but no signs of retained intraocular foreign body. He presented with a total white cataract with brownish deposits on anterior capsule and underwent cataract surgery for same followed by histopathological staining of anterior capsule for iron deposits. This case illustrates the importance of close monitoring of patients with history of trauma or previous penetrating injury to the eye, albeit no intraocular foreign body, as they might develop ocular siderosis at a later stage.

Inadvertent Trypan Blue Posterior Capsule Staining During Cataract Surgery

To report 5 cases of inadvertent posterior capsule staining with trypan blue during phacoemulsification. Retrospective, observational case series. Five cases of posterior capsule staining with trypan blue were identified from cataract surgeries performed at an academic institution. All 5 eyes underwent phacoemulsification with use of iris retractors. The surgical videos from each case were reviewed to better understand the mechanisms and risk factors for posterior capsule staining with trypan blue and techniques to avoid this complication. No eyes had clinical evidence of zonular pathology during their preoperative examination. Only 1 patient reported a possible childhood history of trauma to both eyes. One eye had uveitis, requiring posterior synechialysis. All 5 cases involved the use of iris retractors. No other intraoperative complications occurred, and the intraocular lens was successfully placed in the capsular bag in all cases. All eyes had resolution of posterior capsule staining by postoperative day 8. Inadvertent posterior capsule staining with trypan blue can occur in eyes that appear structurally normal. The use of iris retractors may facilitate posterior capsule staining by allowing the posterior flow of trypan blue under the iris and through the zonules to the posterior capsule. Surgeons should consider techniques to minimize the risk of posterior capsule staining, particularly in cases involving the use of iris retractors.

Characterization of Native Bacillus Thuringiensis (Berliner) Isolates from India

Five native isolates of Bacillus thuringiensis (B.t) were morphologically and biologically characterized. Gram staining, capsule staining, endospore staining and transmission electron microscopy confirmed the B.t. by morphologically. Poly-acrylamide-gel-electrophoresis showed that all the native isolates were producing 130–140 kDa proteins. Chemical characterization revealed that Isolates NSC-1, NSC-3 and COR-4 were fermented maltose while NSC-9 and GUR-5 showed delayed +ve reaction. All selected cultures were catalase +ve, ammonification −ve, citrate utilization +ve, casein hydrolysis +ve, gelatin hydrolysis −ve, indole production −ve and VP test −ve. In the bioassays, five native strains NSC-1, NSC-3, NSC-9, COR-4 and GUR-5 were identified and proved as a novel isolate for controlling Castor hairy caterpillar (Pericalia ricini, Arctidae; Lepidoptera).

Radical distal gastrectomy in laparoscopic and open surgery: is it necessary for pancreatic capsule resection?

To explore the involvement of the pancreatic capsule during radical gastrectomy in gastric cancer. Pancreatic capsule samples were collected from the 83 cases (56 men and 27 women) during open radical gastrectomy and laparoscopic resection between January 2007 and July 2008. RT-PCR and immunohistochemistry were applied for tumor detection. There was a 2-year follow-up; the relationship of the pancreatic capsule involvement, tumor stage and survival rate were evaluated. Results from radical distal gastrectomy were combined with those of gastric cancer pancreatic capsule cleaning; clinical data, pathology, immunohistochemistry and RT-PCR were used to confirm the necessity of pancreatic capsule resection in laparoscopic radical gastrectomy. H&E staining of the pancreatic capsule showed no tumor existence in any of the 83 patients but immunohistochemistry showed CK20 positive cells in 20 patients (33.7%), while RT-PCR detected CK20 mRNA positive cells in 42 patients (50.6%). Cases with stage T1 and T2 were negative for CK20 in both RT-PCR and immunohistochemistry and the few cases with T3 and T4 were also negative in both RT-PCR and immunohistochemistry. The metastasis in the pancreatic capsule correlated mainly with the invasive serous membrane, lymph node metastasis and tumor stage (p<0.05) but not with gender and age (p>0.05). For T1 and T2 stage, there was no evidence of pancreatic capsule metastasis, which may facilitate the decision making of the pancreatic capsule resection during radical distal gastrectomy.

Synthesis, staining properties, and biocompatibility of a new cyanine dye for ILM peeling

The aim of this work is to investigate the biocompatibility and staining properties of DSS: 3,3'-Di-(4-sulfobutyl)-1,1,1',1'-tetramethyl-di-1H-benz[e]indocarbocyanine (DSS). Dye concentrations of 0.5, 0.25, and 0.1% were evaluated (290 and 295 mOsm). Toxicity was assessed using a colorimetric test measuring the inhibition of ARPE 19 cell, human primary RPE cell, and human Müller cell proliferation. Exposure time was 30, 60, 120, and 300 s. Indocyanine green (ICG) (0.5, 0.25, and 0.1%) served as a control. Cells were also illuminated with plain white light (750 mW/cm(2)) for 10 min to assess phototoxic effects. Besides staining of porcine and human lens capsule, internal limiting membrane (ILM)-staining was assessed by applying 0.25 and 0.5% DSS over the macula in two human post-mortem eyes. DSS of 0.25 and 0.1% showed no toxic effect on primary RPE cells and MIO-M1cells, and 0.5, 0.25, and 0.1% for ARPE-19 cells. In MIO-M1cells, 0.5% dye showed a significant reduction of mitochondrial dehydrogenase activity only following an exposure time of 300 s. Following illumination, ICG showed a significantly more pronounced effect on cell viability in primary RPE cells and MIO-M1cells compared to DSS. The absorption maximum is found at 591 nm; the even more bathochromic fluorescence proceeds with a common Stokes' shift where maxima at 620 and 660 nm with a quantum yield of 32% were found. The fluorescence is sufficiently hypsochromic and the fluorescence quantum yield high enough for an easy visual detection. The contrast and staining properties at the ILM were excellent and allowed for a controlled removal of the ILM during surgery. No penetration into deeper retinal layers was noted. Our results indicate that this new cyanine dye DSS may represent an alternative for ILM staining due to its matched absorption concerning visibility and fluorescence qualities as well as its good biocompatibility.

Unintentional staining of the posterior lens capsule with trypan blue dye during phacoemulsification: case report

We report a complication related to the use of trypan blue during capsular staining of the anterior lens capsule during phacoemulsification surgery. In phacoemulsification surgery of the left eye, trypan blue was injected using an air bubble technique. Unintentionally, trypan blue was administered under high pressure, dispersing the dye through the zonules leading to staining of the posterior capsule. This caused a temporary disturbance of visual acuity during the postoperative period.

New cyanine dye for ILM staining

AbstractPurpose To investigate the biocompatibility and staining properties of a new cyanine dye (DSS: 3,3´‐Di‐(4‐sulfobutyl)‐1,1,1´,1´‐tetramethyl‐di‐1H‐benz[e]indocarbocyanine).Methods Dye concentrations of 0.5%, 0.25% and 0.1% were evaluated (osmolarity between 290 and 295 mOsm). Toxicity was assessed using a colorimetric test measuring the inhibition of ARPE 19 cell, human primary RPE cell and human Müller cell proliferation. Exposure time was 30, 60, 120 and 300 seconds. Indocyanine green (ICG) (0.5%, 0.25% and 0.1%) served as a control. Besides staining of porcine and human lens capsule, internal limiting membrane (ILM)‐staining was assessed by applying 0.25% and 0.5% DSS over the macula in two human post‐mortem eyes.Results The dye DSS did not reveal any toxicity on ARPE‐19, primary human RPE cell and human Müller cells proliferation in all concentrations and exposure times investigated. The absorption maximum is found at 591 nm, the even more bathochromic fluorescence proceeds with a common Stokes’ shift where maxima at 620 and 660 nm with a quantum yield of 32% were found. The fluorescence is sufficiently hypsochromic and the fluorescence quantum yield high enough for an easy visual detection. The contrast and staining properties at the ILM was excellent because of matched optical properties and allowed for a controlled removal of the ILM during surgery. No penetration into deeper retinal layers was noted.Conclusion Our results indicate that this new cyanine dye DSS may represent an alternative for ILM staining due to its matched absorption concerning visibility and fluorescence qualities as well as its good biocompatibility. The dye is superior compared with ICG where there is no matching of the UV/Vis spectra.

Anterior lens capsule has more affinity to trypan blue in patients with pseudoexfoliation

Sir, Although not necessarily needed in routine cases, capsule staining (ie, trypan blue) could be especially helpful in pseudoexfoliation syndrome (PEX).1 Conceptually, the presence of the exfoliative material on the anterior lens capsule may interfere with staining capabilities of vital dyes.

Methyl blue and aniline blue versus patent blue and trypan blue as vital dyes in cataract surgery: Capsule staining properties and cytotoxicity to human cultured corneal endothelial cells

To evaluate capsule-staining properties and biocompatibility of the triarylmethane dyes methyl blue and aniline blue compared with patent blue and trypan blue on cultured human corneal endothelial cells. Centre for Ophthalmology, University of Tuebingen, Tuebingen, Germany. Experimental study. Human corneal endothelial cell cultures were harvested from human donor cells and exposed to various concentrations (0.025 to 5.0 mg/mL) of methyl blue, aniline blue, patent blue, and trypan blue. Cytotoxicity was assessed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide test after 24 hours of incubation. Calcein live cell staining was performed at the same time point. The dyes were also used to stain pig lens capsules in vitro by incubating the lenses for 1 minute with 3 concentrations (0.5, 1.5, and 2.5 mg/mL) of dye, after which the staining properties were evaluated. No significant cytotoxicity was detected for patent blue and methyl blue at any tested concentration. However, aniline blue exerted significant cytotoxicity at concentrations of 1.5 mg/mL or higher and trypan blue at 2.5 mg/mL or higher. Capsule staining of the tested triarylmethane dyes was suitable for performing capsulorhexis, but only at higher concentrations than with trypan blue. High concentrations and long incubation times of trypan blue and aniline blue showed significant cytotoxicity to human cultured endothelial cells in contrast to patent blue and methyl blue. All tested dyes were able to stain lens capsules sufficiently for capsulorhexis creation. No author has a financial or proprietary interest in any material or method mentioned.

The T7-Related Pseudomonas putida Phage φ15 Displays Virion-Associated Biofilm Degradation Properties

Formation of a protected biofilm environment is recognized as one of the major causes of the increasing antibiotic resistance development and emphasizes the need to develop alternative antibacterial strategies, like phage therapy. This study investigates the in vitro degradation of single-species Pseudomonas putida biofilms, PpG1 and RD5PR2, by the novel phage ϕ15, a ‘T7-like virus’ with a virion-associated exopolysaccharide (EPS) depolymerase. Phage ϕ15 forms plaques surrounded by growing opaque halo zones, indicative for EPS degradation, on seven out of 53 P. putida strains. The absence of haloes on infection resistant strains suggests that the EPS probably act as a primary bacterial receptor for phage infection. Independent of bacterial strain or biofilm age, a time and dose dependent response of ϕ15-mediated biofilm degradation was observed with generally a maximum biofilm degradation 8 h after addition of the higher phage doses (104 and 106 pfu) and resistance development after 24 h. Biofilm age, an in vivo very variable parameter, reduced markedly phage-mediated degradation of PpG1 biofilms, while degradation of RD5PR2 biofilms and ϕ15 amplification were unaffected. Killing of the planktonic culture occurred in parallel with but was always more pronounced than biofilm degradation, accentuating the need for evaluating phages for therapeutic purposes in biofilm conditions. EPS degrading activity of recombinantly expressed viral tail spike was confirmed by capsule staining. These data suggests that the addition of high initial titers of specifically selected phages with a proper EPS depolymerase are crucial criteria in the development of phage therapy.

Comparative Study of Different Concentration of Trypan Blue for Staining of Capsule in Mature Cataract using RGB Scale in “Adobe Photoshop Image Reader”

Satisfactory capsulorhexis is essential in modern small incision cataract surgery. Visualization of capsule in mature cataract is difficult so staining of capsule with dye is required. Trypan blue is most commonly used dye. Carcinogenic effect of trypan blue is proved in animal study. Aim of this study was to determine minimum concentration of trypan blue that stains the anterior capsule. Methods: The study was conducted in 50 patients with senile mature cataract. Capsular staining was done with different concentration of trypan blue (Group A 0.06%, Group B 0.03%, Group C 0.015%, Group D 0.0075% and Group E 0.00375%) to enhance visualization of capsule for rhexis in cataract surgery. Comparative study of blueness of dye with different concentration was done by RGB scale values in “Adobe Photoshop image reader” Software (Version-7). Results : Trypan blue stains the capsule satisfactorily in group A, B and C. In group D and E capsular staining was not satisfactory. Conclusion: Concentrations of 0.03% and 0.015% were as effective as 0.06% to stain the capsule. Concentration of 0.0075% and 0.00375% could not stain the capsule.

Differential Staining of Bacteria: Capsule Stain

Bacterial capsules are composed of high-molecular-weight polysaccharides and/or polypeptides, and are associated with virulence and biofilm formation. Unfortunately, capsules do not stain well with crystal violet, methylene blue, or other simple stains. This unit describes two methods of capsule staining. The first is a wet-mount method using india ink; the capsule is visualized as a refractile zone surrounding a cell. The second is a direct-staining dry-mount method that precipitates copper sulfate and leaves the capsule as a pale blue zone. Both methods are easily performed within approximately 5 min.

Bilateral electric cataracts: Clinicopathologic report

We report a case with bilateral intumescent electric cataracts, outcomes of cataract surgery with a new technique, and a histopathologic study of the anterior capsule followed by a review of the literature on electric cataracts. The patient had bilateral cataract extraction and posterior chamber intraocular lens implantation, achieving a visual acuity of 20/20. Hematoxylin and eosin staining of the anterior capsule revealed significant scar tissue formation consisting of fibroblast proliferation and hyaloid production over the basement membrane of the anterior capsule. Electric injuries can cause bilateral intumescent cataracts; the outcomes after cataract surgery are excellent provided the fundus and optic nerve examinations are normal. Scar formation over the anterior capsule may disturb lens nutrition, leading to cataract formation.

Capsule staining combined with capsule decompression to facilitate CCC for intumenscent white cataract

目的 探讨晶状体囊染色联合减压撕囊技术在膨胀性白色白内障环形撕囊中的效果及其手术技巧.方法 46例膨胀性白色白内障施行了前囊染色联合减压撕囊技术.采用黏弹剂下晶状体囊染色法.即前房注入healon,用27号钝针头抽取0.5%吲哚青绿溶液0.1mL,自辅助切口紧贴前囊表面将染色剂均匀涂抹在前囊中央直径约6mm的表面.用一次性5mL注射器所带针头,针尖斜面向下在前囊中央刺入前皮质内抽吸液化皮质,前房补充healon,用撕囊镊做连续环形撕囊.结果 经晶状体囊染色及减压处理后,撕囊的可见度及顺应性显著提高,环形撕囊均获成功.未出现任何染色剂以及减压技术的并发症.结论 晶状体囊染色联合减压撕囊法简单易行,值得在临床中应用。

Safety and efficacy of manual small incision cataract surgery for brunescent and black cataracts

To evaluate the safety, visual outcome and complications of manual small incision cataract surgery (MSICS) in the treatment of patients with brunescent and black cataract (BBC). In a non-randomised interventional case series, 102 consecutive patients with BBC underwent cataract extraction by MSICS, with staining of the anterior capsule by trypan blue. Of the 102 eyes with BBC, MSICS was performed through superior scleral tunnel (SST) in 31 eyes (30.4%) and through temporal scleral tunnel (TST) in 71 eyes (69.6%). The main intraoperative complication was posterior capsule rupture in two patients (2.0%). Postoperatively, 20 eyes (19.6%) developed corneal oedema. Mild iritis was seen in six eyes (5.9%) and moderate iritis with fibrin membrane formation occurred in three eyes (2.9%). On the 40th postoperative day, 80 patients (78.4%) achieved uncorrected visual acuity of 6/18 or better, and 99 (97.1%) had best-corrected visual acuity of 6/18 or better. Patients in the SST group had significantly higher postoperative astigmatism compared to those in the TST group (-1.08 D vs -0.72 D, P=0.017). MSICS with trypan blue staining of the anterior capsule is a safe and effective method of cataract extraction for patients with BBC.

A comparative clinical study of anterior capsule staining for capsulohexis in cases of whit cataract with methylene blue and trypan blue

目的 探讨晶状体超声乳化术中改良亚甲蓝染色和台盼蓝染色对连续环形撕囊成功牢及对角膜内皮的影响.方法 67例(67眼)白色白内障(术中无红光反射者)随机分为3组进行前瞻性研究.改良亚甲蓝染色组25例,台盼蓝染色组24例,对照组(前囊不染色)18例.连续环形撕囊后行超声乳化及人工晶状体植入术.比较环形撕囊成功率、术中并发症、术后前房反应情况、视力、眼压、角膜内皮损失率及变异系数.结果 两种方法染色后撕囊成功率及人工晶状体囊袋内植入牢均为100.00%,无后囊破裂,与对照组相比差异有统计学意义;染色组术后视力高于对照组,但差异无统计学意义;3组术后眼压及角膜内皮损失率和变异系数筹异均无统计学意义.结论 改良亚甲监染色和台盼蓝染色均可明显提高连续环形撕囊成功率,降低超声乳化手术并发症,且无明显副作用,前者廉价易得,使用效果与台盼监相同。

Cataract surgery and anterior megalophthalmos: Custom intraocular lens and special considerations

We report a patient who presented with anterior megalophthalmos: corneal diameter of nearly 17.0 mm, anterior chamber depth of 7.0 mm, mild lens subluxation, and nuclear sclerotic cataract. Surgical management consisted of a scleral tunnel incision, capsule staining, a predetermined capsulorhexis size, microcoaxial phacoemulsification with torsional ultrasound, and implantation of a custom IOL to ensure endocapsular fixation. Special consideration must be given to the patient with a very large anterior segment.

Phacoémulsification des cataractes blanches en utilisant le bleu trypan

To study the results of phacoemulsification in eyes with white cataract using trypan blue for capsule staining. Thirty patients who had white cataract were included in this prospective study. They had phacoemulsification and continuous curvilinear capsulorhexis using trypan blue. Intraoperative problems related to capsulorhexis and phacoemulsification as well as postoperative outcomes where analyzed. Postoperative examinations were done at 1 day, 1 week, and 1, 3, 6, and 12 months. The mean follow-up was 10 months. Complet capsulorhexis was achieved in 26 cases. Intraoperative complications included incomplet capsulorhexis in four cases (13.3%) and conversion to a conventional extraction technique in two cases. There were no cases of posterior capsular tear. Twenty-eight eyes (93.3%) had a final best corrected visual acuity at 8/10 or better. Postoperative complications included moderate transient corneal edema in six cases (20%) and iritis in three cases (10%). There were no complications related to the use of trypan blue. Phacoemulsification using trypan blue was safe and effective in managing white cataract and had a high success rate.

Paediatric cataract surgery

Bilateral congenital cataract is the most common cause of treatable childhood blindness. Nuclear cataract is usually present at birth and is non-progressive, while lamellar cataract usually develops later and is progressive. Prompt surgery has to be performed in cases with dense congenital cataract: if nystagmus has developed, the amblyopia is unfortunately irreversible. A treatment regime based on surgery within 2 months of life, combined with prompt optical correction of the aphakia and occlusion therapy with frequent follow-up, have been successful in both unilateral and bilateral cases. The surgery ought to include anterior and posterior capsulorexis in all children at the present time. Intraocular lens implantation has been safely performed below the age of 1 year and has also been successfully performed in bilateral cases. Anterior dry vitrectomy should be performed in preschool children to avoid visual axis opacification. Visual axis opacification is the most common complication found after cataract surgery in children. Secondary glaucoma is by far the most sight-threatening complication and is, unfortunately, common in the newborn so lifelong follow-up is essential in these cases.

Alginate biopolymer production by Azotobacter chroococcum from whey degradation

The potential of three Azotobacter chroococcum strains for whey degradation and alginate production were investigated. After dilution, samples were spread plated on isolation agar and Manitol agar and incubated at 30 °C for 24 h. Microorganisms were screened for their ability to whey degradation and alginate production based on colony morphology, negative and capsule staining, ability to decrease the apparent turbidity of the fermentation broths in batch and semi continuous culture by spectrophotometer assay at 400 nanometer and tensiometer assay. Of the three strains tested for whey degradation, only Azotobacter chroococcum 1723 produced significant apparent growth on whey broth and could decrease about 70 % of turbidity in fermentation broth during 6 days in batch culture. Colonies of this strain was characteristically yellow, large, moucoid and slimy on whey agar than Manitol agar after 24 h at 30 °C. Transmission electron microscopy assay and Carbazole reagent were used to recognize the alginate biopolymer. After optimizing environmental factors such as pH, salt concentration and temperature, this strain was able to produce exopolysaccharide greater than 5 mg/mL. Optimum results were obtained when using whey broth as a fermentation medium without extra salt, temperature at 35 °C and pH 7. Increasing inorganic and organic nitrogen sources (yeast extract and NH 4 NO 3 ) reduced whey degradation at least 30%. Transmission electron microscopy assay showed a net-structured polysaccharide capsule around the cells. Semi-continuous culture results demonstrated that, alginate production as well as whey degradation was decreased (1 mg/mL and 30 %).