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PubMed2006

Analysis of cycloheximide-induced apoptosis in human leukocytes: fluorescence microscopy using annexin V/propidium iodide versus acridin orange/ethidium bromide.

Apoptosis is a highly regulated and programmed cell breakdown process characterized by numerous changes. Since it is implicated in many pathological as well as physiological processes, it is vital to have reliable methods for detecting cell death. In this study, we compared several methods for detecting apoptosis and necrosis in human leukocytes. Apoptosis was induced either by incubating the cells with various doses of cycloheximide (CHX) or by 312 nm UVB irradiation. The methods used for detecting apoptosis were light microscopy (May Grunwald-Giemsa and trypan blue staining), fluorescence microscopy (acridin orange/ethidium bromide and annexin V/propidium iodide staining) and agarose gel electrophoresis of fragmented genomic DNA. Our study showed that CHX-induced apoptosis in cultured peripheral blood mononuclear cells but had no effect on apoptosis in polymorphonuclear cells, so its effect depends on cell type. Evaluation and comparison of the methods for detecting apoptosis showed the following. A Giemsa-stained cytospin allows the main morphological characteristics of necrotic and apoptotic death to be recognized. Trypan blue staining, widely used for estimating cell viability, is valueless for detecting apoptosis. Both fluorescence methods provided reliable and reproducible results and distinguished clearly between subpopulations of apoptotic cells, and were closely intercorrelated. Although applicable to a wide spectrum of cell types, agar electrophoresis of extracted DNA cannot be applied to all cell types and apoptotic conditions. Generally, microscopic examination of acridin orange/ethidium bromide stained cells can be recommended as the most reliable of the methods tested.

Acridine Orange↗
PubMed2005

Comparison of dyes for cataract surgery. Part 2: efficacy of capsule staining in a rabbit model.

PURPOSE: To investigate the efficacy of various dyes for anterior capsule staining to facilitate capsulorhexis during cataract surgery. SETTING: Department of Ophthalmology and Institute of Clinical Medicine, College of Medicine, National Cheng Kung University, Tainan, Taiwan. METHODS: Various concentrations of indocyanine green (ICG), methylene blue (MB), gentian violet (GV), trypan blue (TB), and fluorescein sodium (FS) were used to stain rabbit lenses in vitro. After 1 minute of exposure, the staining of each lens was evaluated using a semiquantitative scoring system in which 0 represented no staining/no contrast between the capsule and cortex and 4 represented excellent staining/remarkable contrast between the stained capsule and unstained cortex. The lowest concentration of dye with a score of 4 was considered the lowest effective concentration for capsulorhexis. RESULTS: Based on our scoring system, the lowest effective concentrations for capsulorhexis were ICG 0.25%, MB 0.10%, GV 0.01%, TB 0.10%, and FS 1.25%. The lowest effective concentrations of the 5 dyes provided comparable contrast. Increased concentrations of dye decreased contrast between the capsule and the cortex and were considered less effective for capsulorhexis. CONCLUSION: Any of the following concentrations of dyes can be used for optimal enhancement of anterior capsule visibility for capsulorhexis: ICG 0.25%, MB 0.10%, GV 0.01%, TB 0.10%, and FS 1.25%.

Animals↗
PubMed1995

Significance of hyaluronic acid for evaluation of hepatic endothelial cell damage after cold preservation/reperfusion.

UNLABELLED: Serum hyaluronic acid (HA) levels during or after liver transplantation can reflect graft viability in animal models. Decreased HA clearance in nonviable grafts seems to reflect hepatic endothelial cell damage induced by cold preservation/reperfusion. Therefore, we examined the relationship between HA clearance and endothelial cell damage, using an isolated perfused rat liver (IPRL) model. The grafts were separated into four groups, according to cold preservation time: minimal storage ( CONTROL: n = 9), 4-hr preservation (n = 9), 5-hr preservation (n = 8), 6-hr preservation (n = 9). After cold storage, grafts were put on the recirculating IPRL system and then reperfused for 120 min with 37 degrees C oxygenated Krebs-Henseleit buffer, containing HA and sodium taurocholate. To examine the function of sinusoidal endothelial cells and hepatocytes, serial measurements of HA, total bile acids, and conventional parameters in the perfusate were taken. After the perfusion, a trypan blue exclusion test was done to assess the microscopic sinusoidal lining cell (SLC) damage. HA clearance from the perfusate showed a preservation time-dependent decrease which clearly distinguished between 4- and 6-hr preserved grafts. Trypan blue uptake ratio of SLC increased in accordance with preservation time. HA clearance at 120 min after reperfusion showed a highly significant correlation with histologically assessed SLC damage. These results suggest that HA is useful to evaluate the extent of endothelial cell damage after cold storage and reperfusion. The significance of HA as a predictor of graft viability is also confirmed.

Animals↗
PubMed1984

Viability of exfoliated colorectal carcinoma cells.

The viability of tumour cells shed into the intestinal lumen was determined in 49 patients with carcinoma of the large bowel. Preoperative colorectal lavage was performed in 19 patients and irrigation of the cut ends of the operative specimen in 30 patients. The resulting cell suspensions were centrifuged on a Nycodenz linear density gradient column so that tumour cells, being larger, were concentrated in a band at the top. In 14 of 19 colorectal lavage cases viable tumour cells were recovered, as assessed by their characteristic morphology and ability to exclude trypan blue. A median of 0.78 X 10(6) viable tumour cells was recovered. The median percentage cell viability in the suspension was 92, i.e. 8 per cent of the tumour cells were dead (stained with trypan blue). In eight specimens viability was confirmed by the ability of tumour cells to hydrolyse fluorescein diacetate. In 17 of 30 proximal resection margin irrigations a median of 0.55 X 10(5) viable tumour cells was recovered, with a median percentage viability of 92.5. In 15 specimens the neoplastic cells showed fluorescence. In 21 of 25 distal resection margin irrigations a median of 1.92 X 10(5) viable tumour cells was recovered with a median percentage cell viability of 79.3, and fluorescence was observed in all specimens. The number of viable tumour cells did not correlate with the stage, differentiation, diameter or fixity of the tumour. However, the number of tumour cells recovered from the distal resection margin was inversely related to the distance of the tumour from that margin (Rank Difference Coefficient R = -0.6). Thus viable exfoliated tumour cells were demonstrated in 52 of 74 specimens (70 per cent). Their presence in large numbers at the site of intestinal anastomoses supports a potential role in the aetiology of suture-line recurrence.

Cell Count↗
PubMed1993

Fasting enhances the effects of anoxia on ATP, Cai2+ and cell injury in isolated rat hepatocytes.

The effect of fasting and anoxia on the intracellular concentration of ATP, Na+, Ca2+, Mg2+, and H+ was studied in isolated perfused rat hepatocytes. ATP and intracellular Mg2+ were measured by 31P-NMR spectroscopy, cytosolic free calcium was measured with aequorin, intracellular Na+ with SBFI, intracellular pH with BCECF, lactic dehydrogenase by NADH absorbance. In hepatocytes from fasted rats, intracellular ATP was depressed 52% (P < 0.001), Nai+ was increased 70% from 16.9 to 27.7 mM (P < 0.02), and Cai2+ was increased 79% from 137 to 245 nM (P < 0.05) when compared to fed rats. Mgi2+ and pHi were unchanged. During anoxia, ATP and the cell phosphorylation potential decreased 90% to practically the same low levels in both fed and fasted groups. On the other hand, in hepatocytes from fasted animals, Cai2+ increased faster and to significantly higher levels than in hepatocytes from fed rats: Cai2+ reached 2.19 microM in 10 min compared to 1.45 microM in 1 h, respectively (P < 0.05). Cell injury assessed by LDH release and trypan blue exclusion also occurred earlier and was more severe in hepatocytes from fasted rats. Fructose and Ca(2+)-free perfusion reduced the rise in Cai2+, abolished LDH release and significantly improved the cell viability measured by Trypan blue exclusion. The data demonstrate that fasting decreases the hepatocytes energy potential and increases Nai+ and Cai2+ which are inversely related to the cell energy potential. Consequently, in hepatocytes isolated from fasted rats, the increase in Cai2+ and the resulting cell injury evoked by anoxia occur earlier and are more severe than in fed rats. These results suggest that Ca2+ plays a crucial role in the development of anoxic cell injury.

Adenosine Triphosphate↗
PubMed1991

Selective staining methods for cartilage of rat fetal specimens previously treated with alizarin red S.

A convenient method for staining cartilage with several basic stains after alizarin red S staining of bone was investigated in rat fetuses. It was found that bromophenol blue was useful and effective for staining of the margin and center areas of cartilage, even in specimens stored in glycerin for over 10 years. The specimens were washed in running tap water for 1 hr, and subsequently were immersed in water or in 70% ethanol at pH 4 for 1 hr or longer. The specimens were then stained with 0.005% bromophenol blue in 40% ethanol adjusted to pH 4 for 2 hr, or with 0.001% bromophenol blue in 40% ethanol adjusted to pH 4 for 24 hr. Furthermore, the bromophenol blue stain color actually faded when the specimens were immersed in water or in 70% ethanol at pH 8. Descending order of the stain-effective action on fetal rat cartilage for the basic stains tested was bromophenol blue, aniline blue, Evans blue, methyl violet, trypan blue, and water blue.

Animals↗
PubMed1995

Microcystin uptake and inhibition of protein phosphatases: effects of chemoprotectants and self-inhibition in relation to known hepatic transporters.

The microcystins (Mcyst) are cyclic peptide hepatotoxins produced by cyanobacteria. They are chemically very stable and represent a public health threat when they occur in water bodies used for human consumption. Mice injected ip with Mcyst (LD50 50-100 micrograms/kg) accumulate Mcyst in the liver and die within 2-4 hr with massive intrahepatic hemorrhage. Pretreatment of mice with cyclosporin A (CP), rifamycin (Rif), trypan blue (TB), and trypan red (TR) protected the animals from a lethal dose of the toxin. The studies reported here using the freshly isolated rat hepatocyte model were undertaken in order (1) to evaluate the contribution of Mcyst transport in hepatocytes to the mechanism of chemoprotection for Mcyst in vivo toxicity by CP, Rif, TB, and TR and (2) to better characterize the hepatic Mcyst transporter in this model and determine its relationship to other bile acid/organic anion transporters that have already been fully described. Incubations with 125I-Mcyst were used to measure Mcyst uptake and accumulation in hepatocytes. It has been shown that at the cellular level Mcyst binds to and inhibits protein phosphatases 1 and 2A (PP) at nanomolar concentrations. Lethal doses of Mcyst in mice resulted in rapid profound inhibition of hepatic PP activity. PP activity was also inhibited in hepatocytes incubated with 100-500 nM Mcyst. PP inhibition in these studies was used as a marker of metabolic effects of the toxin. The chemoprotectants CP (5 microM), Rif (50 microM), TR (20 microM), and TB (20 microM) decreased accumulation of Mcyst (320 nM, a toxic concentration) after 30 min incubation to 37, 26, 30, and 66%, respectively, of that of cells treated with Mcyst only. Inhibition of PP activity in these cells was decreased. Inhibition of PP activity in hepatocytes was also decreased by known inhibitors of Mcyst transport: 50 microM of the bile acids cholate and taurocholate (TC) and 50 microM sulfobromophthalein. For all compounds tested the amount of Mcyst accumulated in the hepatocytes correlated qualitatively with the extent of PP inhibition. From these results it can be concluded that inhibition of Mcyst uptake by hepatocytes is the most likely mechanism of chemoprotection for Mcyst in vivo toxicity for TR, TB, CP, and Rif. Uptake of Mcyst was unaffected by changes in the ionic composition of the uptake buffer but was significantly decreased when PP activity of hepatocytes was inhibited by preincubation with Mcyst.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1992

Suramin prevents import of acyl-CoA oxidase into rat liver peroxisomes.

The inhibitory effect of suramin on the import of [35S]acyl-CoA oxidase into purified rat liver peroxisomes was investigated in vitro. The import of acyl-CoA oxidase was inhibited completely by 10 microM suramin, whilst the latency of catalase remained unchanged. The important value decreased 60% by pretreatment of peroxisomes with 10 microM suramin, but it did not decrease by pretreatment of translation products. Polysulfonate compounds which have two clusters of negative charges, such as Cibacron blue F3GA and Trypan blue, as well as suramin, inhibited the import, whilst mono- and disulfonate compounds did not.

Acyl-CoA Oxidase↗
PubMed2001

Effects of 5-aminolaevulinic acid on human ovarian cancer cells and human vascular endothelial cells in vitro.

Results are reported on the cellular effects and the sensitivity of cultured tumor epithelial cells (TEC) derived from human ovarian cystadenocarcinoma and human umbilical vein-derived endothelial cells (HUVEC) to exogenous 5-aminolaevulinic acid (ALA) and ALA-induced photodynamic therapy (PDT). Cellular alterations and PDT efficiency were evaluated using colorimetric thiazolyl blue (MTT) assay, trypan blue exclusion assay, electron microscopy, and gel electrophoresis. ALA-induced protoporphyrin IX (PpIX) accumulation in TEC was associated with a concentration and time-dependent significant decrease in mitochondrial activity, increase in cell membrane permeability, and dark toxicity. Maximum PpIX loaded TEC demonstrated a high sensitivity to PDT. Neither cellular alterations nor PDT effects were observed in HUVEC under identical experimental conditions. These results indicate a potential clinical value for the use of ALA-mediated PDT to treat minimal residual disease in mucinous ovarian carcinoma. In addition, the ALA-induced PpIX cytotoxicity may be exported to a new chemotherapeutic regimen via a conventionally viewed photochemotherapeutic agent.

Aminolevulinic Acid↗
PubMed1995

Melatonin protects neurons from singlet oxygen-induced apoptosis.

Singlet oxygen (O2[1 delta g]) is a very reactive molecule that can be produced by living cells and may contribute to cytotoxicity. The pineal hormone melatonin has been reported to possess potent antioxidant activity, and to be capable of scavenging O2(1 delta g). We investigated whether melatonin might reduce the neurotoxic action of O2(1 delta g). The cytotoxic effect of singlet oxygen was studied in primary cultures of cerebellar granule neurons pretreated with a photosensitive dye, rose bengal, and exposed to light--a procedure that generates O2(1 delta g). We found that this procedure triggers neuronal death, which is preceded by mitochondrial impairment (assayed by the rate of the reduction of MTT, 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide, into formazan), and by DNA fragmentation--a marker of apoptosis. DNA fragmentation was determined in situ by terminal deoxynucleotidyl transferase assay; cell death was assayed with 0.4% trypan blue solution--viable cells with an intact membrane are not permeable to trypan blue; dead cells are, and thus, they are stained blue. Neuroprotection was obtained with the pineal hormone melatonin. In a cell-free system, melatonin also protected the enzyme creatine kinase (EC 2.7.3.2) from the rose bengal-induced injury. The results suggest that melatonin might counteract the cytotoxic action of singlet oxygen. Further studies are needed to clarify the exact role singlet oxygen and melatonin might play in neurodegenerative diseases.

Animals↗
PubMed1974

Streptolysin O inhibition of neutrophil chemotaxis and mobility: nonimmune phenomenon with species specificity.

The effects of streptolysin O (SO) (1 to 4 hemolytic units) on the mobility of neutrophilic leukocytes from humans, baboons, sheep, and rabbits were compared. After SO treatment, chemotaxis and random mobility of human neutrophils were markedly suppressed, baboon and sheep neutrophils were partially suppressed, and rabbit neutrophils were unaffected and demonstrated normal chemotaxis and mobility. The amounts of SO used in the mobility studies caused no leukocyte lysis or trypan blue uptake by human, baboon, or sheep cells, and minimal lysis or trypan blue uptake by rabbit cells. The possible involvement of immune mediators in the observed inhibition of human neutrophils was considered and excluded by the following studies. White blood cells from humans with humoral or cellular immune deficiencies responded in a manner similar to normal human cells; supernatant solutions from SO-treated human white blood cells did not contain a chemotactic suppressor; preincubation of SO with cholesterol (an inhibitor of SO hemolytic activity) caused loss of the chemotactic suppressive effect of the toxin on human leukocytes; and leukocytes from rabbits preimmunized with SO remained refractory to chemotactic suppression.

Agammaglobulinemia↗
PubMed2001

Traceability of aquatic animals.

Effective methods of traceability are urgently required for use in research as well as in different types of aquaculture operations and to control trade in aquatic animals and products. In regard to the marking of fish, many different tagging methods have been described and the method to be used depends on the purpose and need for tagging. In contrast, for molluscs and crustaceans, only a few methods of marking such animals have been described, due to the practical difficulties. The authors first describe the different methods for tracing fish and fishery products, by means of external tags, such as Floy tags, Carlin tags and passive integrated transponder tags; chemical marking using inorganic substances such as silver nitrate or potassium nitrate, pigments, oxytetracycline, etc.; and several different types of electronic devices in which basic information such as the strain of fish, farm of origin or weight can be stored. Genetic traceability using deoxyribonucleic acid profiling is developing quite rapidly for cultured brood stocks and wild populations. This technique may be used with very high degrees of confidence to assign to or exclude animals or products from their claimed origin, paternity or strain, and may be used as evidence in court proceedings. The second section of this paper describes the traceability of live molluscs for restocking and for human consumption. In these applications, genetic markers have been demonstrated to be suitable. Mechanical tagging on a small scale for research purposes has also been used. Otherwise, the only means of tracing live molluscs are the movement documents and the labelling on boxes that certifies the origin of the commodity. The third section describes the methods available for tracing live and dead crustaceans. A large variety of physical tagging methods for decapod crustaceans is described, such as the injection of biological stains (fast green, Niagara sky blue, trypan red and blue) and external tags such as coloured streamer tags, wire tags and a variety of anchor tags. Furthermore, a number of different internal coding methods, such as the coded micro-wire tags and injected elastomer tags are discussed in detail. As is the case for fish, genetic molecular techniques are also applied in population studies of crustaceans; some of the molecular genetic methods are described. Prawns for human consumption are most frequently packed whole or as tails after the necessary sorting, washing and freezing and the only way of performing a traceback is through documents relating to movement, invoices, health certificates and labelling of the boxes. The minimum requirements for labelling would be the content of the packages, i.e. species, quantity, identification of the manufacturer (name and address), packing place, importer/exporter or vendor of the product, in addition to the loading bill number.

Animal Identification Systems↗
PubMed1991

Decontamination of aqueous solutions of biological stains.

Aqueous solutions of a number of biological stains were completely decontaminated to the limit of detection using Amberlite resins. Amberlite XAD-16 was the most generally applicable resin but Amberlite XAD-2, Amberlite XAD-4, and Amberlite XAD-7 could be used to decontaminate some solutions. Solutions of acridine orange, alcian blue 8GX, alizarin red S, azure A, azure B, Congo red, cresyl violet acetate, crystal violet, eosin B, erythrosin B, ethidium bromide, Janus green B, methylene blue, neutral red, nigrosin, orcein, propidium iodide, rose Bengal, safranine O, toluidine blue O, and trypan blue could be completely decontaminated to the limit of detection and solutions of eosin Y and Giemsa stain were decontaminated to very low levels (less than 0.02 ppm) using Amberlite XAD-16. Reaction times varied from 10 min to 18 hr. Up to 500 ml of a 100 micrograms/ml solution could be decontaminated per gram of Amberlite XAD-16. Fourteen of the 23 stains tested were found to be mutagenic to Salmonella typhimurium. None of the completely decontaminated solutions were found to be mutagenic.

Acrylic Resins↗
PubMed1994

Removal of biological stains from aqueous solution using a flow-through decontamination procedure.

Chromatography columns filled with Amberlite XAD-16 were used to decontaminate, using a continuous flow-through procedure, aqueous solutions of the following biological stains: acridine orange, alcian blue 8GX, alizarin red S, azure A, azure B, brilliant blue G, brilliant blue R, Congo red, cresyl violet acetate, crystal violet, eosin B, eosin Y, erythrosin B, ethidium bromide, Giemsa stain, Janus green B, methylene blue, neutral red, nigrosin, orcein, propidium iodide, rose Bengal, safranine O, toluidine blue O, and trypan blue. Adsorption was most efficient for stains of lower molecular weight (< 600). Adsorption of stain increased as the flow rate decreased; column diameter had little effect on adsorption. Adsorption of stain was greatest when finely ground resin was used, but if the resin particles were too small, column clogging occurred. Limited grinding of the resin gave increased adsorption while retaining good flow characteristics. Amberlite XAD-16 saturated with methylene blue was regenerated to its initial adsorption capacity by passing methanol through the column. The technique described provides an economical, rapid means of removing stains from aqueous solution.

Animals↗
PubMed1989

Assessment of human natural killer and lymphokine-activated killer cell cytotoxicity against Toxoplasma gondii trophozoites and brain cysts.

Because previous work has suggested that NK cells may be important in host resistance against the intracellular parasite Toxoplasma gondii we examined whether human NK cells and lymphokine-activated killer (LAK) cells have activity against trophozoites and cysts of this organism in vitro. A method to radiolabel Toxoplasma trophozoites with 51Cr was developed and direct cytotoxic activity was determined by using modifications of the standard 51Cr release assay. Viability of 51Cr-labeled trophozoites assessed by both methylene blue staining and trypan blue exclusion was greater than 90%. Significantly more 51Cr was released by anti-Toxoplasma antibody and C than by antibody in the absence of C. Incubation of trophozoites with freshly isolated human NK cells or NK cells activated with either rIL-2 or rIFN-alpha did not result in significant release of 51Cr (specific lysis was 0 to 2.3%). In contrast, the average specific lysis of radiolabeled trophozoites by LAK cells was significant (specific lysis was 7.8% +/- 1.1, p less than 0.01). In a series of separate experiments, preincubation of radiolabeled trophozoites with heat-inactivated normal or Toxoplasma antibody-positive human serum increased the cytotoxicity of LAK cells from a mean specific lysis of 15% +/- 4.5 to 39% +/- 8.5, respectively (p less than 0.05), as assessed by 51Cr release. Because previous work has shown that radioisotope release from parasites may be nonspecific, separate experiments were performed to determine the cytotoxicity of LAK cells against antibody-coated trophozoites by using ethidium bromide-acridine orange staining to assess effector cell damage. LAK cells had a mean specific lysis of 51% against antibody-coated trophozoites by ethidium bromide-acridine orange staining. Preincubation with heat-inactivated Toxoplasma-antibody positive human serum did not increase activity of rIL-2-activated NK cells against 51CR-labeled trophozoites. Neither human NK cells (freshly isolated or activated by rIL-2 or rIFN-alpha) nor LAK cells were cytotoxic for purified preparations of cysts of Toxoplasma isolated from the brains of chronically infected mice.

Acridine Orange↗
PubMed1978

Can steroid reduce endothelial damage in stored corneas? Effect on cell viability and ultrastructure.

We studied the effect of steroid on the viability and integrity of the endothelium of variously stored corneas of different species of animals and man. Paired corneas of rodents and humans were used,--one cornea (experimental) being treated with steroid and the other (control) not. Rat and guinea pig corneas were kept at 37 degrees C for one hour in phosphate buffered saline with or without hydrocortisone 21-sodium succinate (10-6M). Human corneas were kept at 4 degrees C for 4 days in M-K medium with or without hydrocortisone 21-sodium succinate (10-6M). The viability of the endothelia (rat and guinea pig) was tested with para Nitro Blue Tetrazolium and trypan blue dye exclusion tests. The ultrastructural changes in the control and experimental endothelial (guinea pig and human) were examined by scanning electron microscopy. Viable cells P less than 0.001) were more numerous and ultrastructural alteration less in the endothelia of the steroid treated corneas compared to the controls.

Animals↗
PubMed1975

[Utilization of rubidium 86 for humoral cytotoxicity tracing of sarcoma 180].

86Rb has been used as a tracer for Sarcoma 180 ascitic cells in a humoral cytotoxicity system, and compared with the trypan blue dye exclusion method. In as little as five minutes after contact with antiserum and complement, about 90 p. cent of the isotope has gone from the cells, with a significant difference between controls and experimentals. There is a good correlation between 86Rb and trypan blue dye exclusion results. The only pitfall is a relative fragility of cells during the centrifugation stages of the two washings.

Animals↗
PubMedDate not supplied

A comparison of the effectiveness of four biostains in enhancing visualization of the vitreous.

PURPOSE: To compare the effectiveness of four biostains in vitreous staining as a means to facilitate clean anterior vitrectomy. MATERIALS AND METHODS: Four biostains (fluorescein, indocyanine green, trypan blue, and triamcinolone acetonide) were used to stain the transparent vitreous of human cadaver and monkey cadaver eyes. Results were compared and recorded by digital photography and videography. RESULTS: Of the four biostains, triamcinolone acetonide best highlighted the vitreous. Although fluorescein, indocyanine green, and trypan blue also produced acceptable vitreous staining, the surrounding ocular structures were slightly stained as well. CONCLUSION: All four biostains enhanced visualization of the vitreous. However, triamcinolone acetonide best highlighted the vitreous.

Animals↗