Abstract：
:A duplex fluorescence assay to assess the viability of cells cultured in multi-well plates is described, which can be carried out in the original culture plate using a plate reader, without exchanges of culture or assay medium, or transfer of cells or cell supernatant. The method uses freshly prepared reagents and does not rely on a proprietary, commercially supplied kit. Following experimental treatment, calcein acetoxymethyl ester (CaAM) is added to each well of cultured cells; after 30 min, the fluorescence intensity (emission λmax ∼ 530 nm) is measured. The signal is due to formation of calcein, which is produced from CaAM by action of esterase activity found in intact live cells. Since live cells may express plasma membrane multidrug transport proteins, especially of the ABC transporter family, the CaAM incubation is carried out in the presence of an inhibitor of this efflux process, thereby improving the dynamic range of the assay. Next, SYTOX® Orange (SO) is added to the culture wells, and, after a 30-min incubation, fluorescence intensity (emission λmax ∼ 590 nm) is measured again. SO is excluded from cells that have an intact plasma membrane, but penetrates dead/dying cells and can diffuse into the nucleus, where it binds to and forms a fluorescent complex with DNA. The CaAM already added to the wells causes no interference with the latter fluorescent signal. At the conclusion of the duplex assay, both live and dead cells remain in the culture wells and can be documented by digital imaging to demonstrate correlation of cellular morphology with the assay output. Two examples of the application of this method are provided, using cytotoxic compounds having different mechanisms of action.
journal_name
Exp Eye Resjournal_title
Experimental eye researchauthors
Pfeffer BA,Fliesler SJdoi
10.1016/j.exer.2017.05.011subject
Has Abstractpub_date
2017-08-01 00:00:00pages
17-29eissn
0014-4835issn
1096-0007pii
S0014-4835(17)30112-4journal_volume
161pub_type
杂志文章abstract：:AII amacrine cells are critical interneurons in the rod pathway of mammalian retina, active primarily in dim lighting conditions. Melatonin, a neuromodulator produced at night in the retina, is believed to induce retinal adaptation to dim lighting conditions in most vertebrate species examined to date, including human...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/s0014-4835(03)00123-4
更新日期：2003-09-01 00:00:00
abstract：:Situated at the junction between inner and outer segment, the connecting cilium of retinal photoreceptors supports regulated transport of molecules that function distally, while restricting diffusion of membrane proteins from one plasmalemmal domain to the other. Both functions are thought to be performed by a group o...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1006/exer.1996.0261
更新日期：1997-06-01 00:00:00
abstract：:Preeclampsia (PE) is a hypertensive complication of pregnancy. Its cause is still unknown and it could be a risk factor for future ophthalmic problems. Retinal vascular bed alterations have been described as a consequence of PE, suggesting a retinopathy. Factors related to angiogenesis and vascular permeability, such ...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/j.exer.2020.107981
更新日期：2020-04-01 00:00:00
abstract：:Simvastatin is a cholesterol-lowering drug which exhibits numerous pleiotropic effects including anti-cancer activity. Yet, the anti-cancer effects in choroidal melanoma remain poorly characterized. Therefore, in this study, we investigated the effects of simvastatin on OCM-1 cells growth, apoptosis and cycle. Simvast...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/j.exer.2013.05.013
更新日期：2013-08-01 00:00:00
abstract：:[3H]-retinol binding to cellular retinol-binding protein (CRBP) increased more than five-fold in developing chicken corneal epithelium at 14 days of incubation, the time when the initial increase in corneal transparency occurs in the developing embryo. In contrast, [3H]-retinoic acid binding to cellular retinoic acid-...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/0014-4835(83)90026-x
更新日期：1983-11-01 00:00:00
abstract：:The P23H-1 rat strain carries a rhodopsin mutation frequently found in retinitis pigmentosa patients. We investigated the progressive degeneration of the inner retina in this strain, focussing on retinal ganglion cells (RGCs) fate. Our data show that photoreceptor death commences in the ventral retina, spreading to th...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/j.exer.2010.10.003
更新日期：2010-12-01 00:00:00
abstract：:A dark reaction is known to occur in retinal extracts of the gecko (Gekko gekko), in which the natural 11-cis-chromophore of the 521-pigment is apparently replaced by adding 9-cis-retinal to form the 9-cis-photopigment. With chloride-deficient extracts the reaction involves some 70% of the 521-pigment. Anions like nit...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/0014-4835(89)90074-2
更新日期：1989-07-01 00:00:00
abstract：:Immunocytochemistry with a monoclonal antibody against the 28-kDa calcium-binding protein (28-kDa CaBP) was used to study horizontal cells in both congenic control (RCS-rdy+) and dystrophic (RCS) 3-month-old rat retina. We found that horizontal cells in the RCS-rdy+ rat retinas were distributed evenly from the centre ...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1006/exer.1993.1109
更新日期：1993-08-01 00:00:00
abstract：:The development of a synthetic material allowing increased cellular adhesion and ingrowth would improve keratoprosthetic devices and requires an understanding of cell colonization processes. Interlamellar implantation of hydrophobic synthetic material may lead to poor corneal nutrition with subsequent necrosis. The pr...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1006/exer.1994.1193
更新日期：1994-01-01 00:00:00
abstract：:Diabetic retinopathy is a complication of diabetes and a leading cause of vision loss among working-age adults. To assess whether the Wistar rat with Streptozotocin (STZ)-induced diabetes is a suitable animal model of human proliferative diabetic retinopathy we evaluated the vascular changes to assess the diabetic ret...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/j.exer.2019.04.025
更新日期：2019-07-01 00:00:00
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journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/j.exer.2013.10.019
更新日期：2014-01-01 00:00:00
abstract：:A method for the isolation and purification of human ocular mucin from the brief saline extract of human ocular mucus is reported. Initial purification of ocular mucin was achieved by sequential chromatography of the saline-soluble mucus extract from an individual donor's mucus pool on columns of Sephadex G-50 and Sep...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/0014-4835(88)90002-4
更新日期：1988-08-01 00:00:00
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journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/s0014-4835(87)80178-1
更新日期：1987-03-01 00:00:00
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journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/s0014-4835(86)80030-6
更新日期：1986-10-01 00:00:00
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journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/j.exer.2005.01.004
更新日期：2005-07-01 00:00:00
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journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1006/exer.1997.0371
更新日期：1997-10-01 00:00:00
abstract：:As a potential drug for the prevention of secondary cataract formation (SCF), we investigated the effect of Aclacinomycin A (ACM) on the growth of cultures of porcine lens epithelial cells in vitro. ACM is an anthracycline that has been used in the treatment of acute myeloid leukemia in the human for many years. It ha...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：
更新日期：1998-01-01 00:00:00
abstract：:SV40 human corneal epithelial cell (HCEC) populations are readily used as a substitute for primary corneal epithelial cells that are difficult to maintain in vitro. To initiate cell-cycle experiments with the SV40-HCEC cells, two separate methods of cell synchronization were compared including serum starvation and ste...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/j.exer.2005.11.007
更新日期：2006-07-01 00:00:00
abstract：:Metabolomics studies in the retina and retinal pigment epithelium (RPE) in animal models or postmortem donors are essential to understanding the retinal metabolism and to revealing the underlying mechanisms of retinal degenerative diseases. We have studied how different methods of euthanasia (CO2 or cervical dislocati...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/j.exer.2018.05.032
更新日期：2018-09-01 00:00:00
abstract：:This study investigated the changes in pH during retinal artery occlusion by means of extracellular H+ concentration ([H+]o) measurements in the retina under both air and 100% O2 ventilation. Occlusion was produced in intact anesthetised cats by pressing with a probe onto a retinal artery. [H+]o profiles were recorded...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/j.exer.2004.11.002
更新日期：2005-04-01 00:00:00
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journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/0014-4835(83)90009-x
更新日期：1983-02-01 00:00:00
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journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/j.exer.2011.06.001
更新日期：2011-10-01 00:00:00
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journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/j.exer.2018.10.002
更新日期：2019-02-01 00:00:00
abstract：:The differentiation of rat lens epithelial cells to fibre cells can be mimicked using lens epithelial explants, which differentiate in vitro when exposed to fibroblast growth factor (FGF). A previous study demonstrated that FGF is required only for initiation of differentiation: once induced by FGF, differentiation ca...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1006/exer.1998.0534
更新日期：1998-10-01 00:00:00
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journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/j.exer.2014.07.004
更新日期：2014-10-01 00:00:00
abstract：:It has long been assumed that unstimulated tears are more thoroughly equilibrated with epithelial secretions than stimulated tears, since they are in contact with tarsal, bulbar and corneal surfaces for longer. It was also believed from results with model solutions that soluble mucin is responsible for the observed su...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1006/exer.1998.0600
更新日期：1999-02-01 00:00:00
abstract：:Previous evidence has shown clearly that sugar cataract formation results from unusually high intracellular levels of polyol. Documentation of polyol-related histological changes in the cortical fiber cells of the equatorial zone has been extensive. However, little attention has been given to the early changes in the ...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/0014-4835(90)90107-6
更新日期：1990-06-01 00:00:00
abstract：:All aqueous humor draining through the conventional outflow pathway must cross the endothelium of Schlemm's canal (SC), likely by passing through micron-sized transendothelial pores. SC pores are non-uniformly distributed along the inner wall endothelium, but it is unclear how the distribution of pores relates to the ...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/j.exer.2014.11.008
更新日期：2015-01-01 00:00:00
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journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/j.exer.2020.107936
更新日期：2020-03-01 00:00:00
abstract：:Exposure of the rabbit lens, in vitro, to 10(-3) M-hydrogen peroxide resulted in an increase of the lens sodium, calcium and water content together with a decrease of the lens potassium content. No such changes were observed in the presence of 10(-4) or 10(-5) M-hydrogen peroxide. 10(-3) M-hydrogen peroxide was observ...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/0014-4835(83)90148-3
更新日期：1983-07-01 00:00:00
