An Easy and Inexpensive Method for Quantitative Analysis of Endothelial Damage by Using Vital Dye Staining and Adobe Photoshop Software

Saad, Hisham A; Terry, Mark A.; Shamie, Neda; Chen, Edwin S.; Friend, Daniel F; Holiman, Jeffrey D.; Stoeger, Christopher G.

doi:10.1097/ico.0b013e3181705ca2

Cited by 68 publications

(73 citation statements)

References 20 publications

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“…Among the techniques used to record and later analyze images of the corneal endothelium there are specular microscopy, scanning electron microscopy and optical microscopy after staining with vital dyes (SAAD et al, 2008;RUGGERI, 2010). Using images obtained by specular microscopy, it is possible to examine the corneal endothelium and obtain data related to endothelial cell density and morphology.…”

Section: Discussionmentioning

confidence: 99%

“…This technique has been used to quantify endothelial parameters in humans and other animal species including pigs (TAMAYO-ARANGO et al, 2009), rabbits (SAILSTAD & PEIFFER, 1981;OJEDA et al, 2001), dogs (GWIN et al, 1982;PIGATTO et al, 2006;RODRIGUES et al, 2006), horses (ANDREW et al, 2001;LEDBETTER & SCARLETT, 2009), llamas and alpacas (ANDREW et al, 2002), chinchillas (BERCHT et al, 2015), and cats (FRANZEN et al, 2010), among others. The high cost of specular microscopes and the difficulty in obtaining good images in injured endothelial areas are common challenges or barriers to the use of this technique (ANDREW et al, 2001;PIGATTO et al, 2005a;SAAD et al, 2008). SEM has been widely used to compare the endothelial ultrastructure of vertebrates and to evaluate the effects of medications, chemicals, or surgical procedures on the endothelium (COLLIN & COLLIN, 1998;PIGATTO et al, 2004;PIGATTO et al, 2005a;PIGATTO et al, 2005b;PIGATTO et al, 2009).…”

Section: Discussionmentioning

confidence: 99%

“…Dual staining of the corneal endothelium with vital dyes using 0.25% trypan blue and 0.2% alizarin red is the most common method for determining viable and nonviable cells in areas where cells cannot be identified using specular microscopy (SPERLING, 1977;TAYLOR & HUNT, 1981). Vital staining with alizarin red and optical microscopy is a rapid, simple, and inexpensive method for obtaining images for morphological and morphometric evaluation of the endothelium in humans and animals (SAAD et al, 2008). In the present study, using an optical microscope was possible to obtain clear images of the endothelial mosaic in all regions of the cornea.…”

Section: Discussionmentioning

confidence: 99%

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Morphology of endothelial cells from different regions of the equine cornea

et al. 2016

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Morphology of endothelial cells from different regions of the equine cornea

et al. 2016

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“…Adobe Photoshop® is a widely available, low cost software, and its use makes this technology available and reproducible to other centers (16,17,(25)(26)(27). The gene that codifies CA-IX is regulated by HIF-1α expression, and HIF-1α is controlled by the VHL protein.…”

Section: Discussionmentioning

confidence: 99%

Carbonic Anhydrase IX is Not a Predictor of Outcomes in Non-Metastatic Clear Cell Renal Cell Carcinoma - A Digital Analysis of Tissue Microarray

et al. 2013

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Introduction:The knowledge about the molecular biology of clear cell renal cell carcinoma (ccRCC) is evolving, and Carbonic Anhydrase type IX (CA-IX) has emerged as a potential prognostic marker in this challenging disease. However, most of the literature about CA-IX on ccRCC comes from series on metastatic cancer, with a lack of series on non-metastatic cancer. The objective is to evaluate the expression of CA-IX in a cohort of non-metastatic ccRCC, correlating with 1) overall survival, and 2) with established prognostic parameters (T stage, tumor size, Fuhrman nuclear grade, microvascular invasion and peri-renal fat invasion). Materials and Methods: This is a retrospective cohort study. We evaluated 95 patients with non-metastatic clear cell renal cell carcinoma, as to the expression of CA-IX. The analyzed parameters where: overall survival (OS), TNM stage, tumor size (TS), Fuhrman nuclear grade (FNG), microvascular invasion (MVI), peri-renal fat invasion (PFI). We utilized a custom built tissue microarray, and the immunoexpression was digitally quantified using the Photoshop® software. Results: The mean follow-up time was 7.9 years (range 1.9 to 19.5 years).The analysis of CA-IX expression against the selected prognostic parameters showed no correlation.

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“…Insult to this monolayer, such as surgical trauma or intracameral injection, poses a risk for permanent harmful effects on the corneal endothelium (PIGATTO et al, 2008;SAAD et al, 2008). In cases of blindness due to cataract, surgical removal of the lens is the standard management.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of equine corneal endothelium after exposure to 0.5% indocyanine green - in vitro study

Silva

Andrade

Faganello

et al. 2018

SCA

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The purpose of the study was to investigate whether indocyanine green (ICG) dye damages the corneal endothelium of horses. Twenty-four corneas of 12 healthy equines, males or females, of different ages were used in this study. Only eyes with no ocular findings were used. Randomly, one eye was included in the treatment group and one in the control group. The eyes of the treatment group were exposed for 1 minute to dye ICG 0.5%. After that the endothelium of all eyes was stained with trypan blue and alizarin red S and analyzed and photographed under an optical microscope. Areas with damaged endothelial cells were manually measured and quantified using software for morphometric analysis and expressed as a percentage of cell damage. In all eyes examined areas of cell damage were observed in both corneas of the control group and the treatment group. The mean endothelial damage was 0.8 ± 0.37% in the treatment group and 0.97 ± 0.39% in the control. The Qui-square test stated that treatment and control group were not different. The ICG 0.5% did not cause acute damage to equine corneal endothelium. Key words: Cataract. Indocyanine green. Corneal endothelium. Alizarin red. ResumoO objetivo do estudo foi investigar se indocianina verde (ICG) induz dano nas células do endotélio da córnea de equinos. Vinte e quatro córneas de 12 equinos saudáveis, machos ou fêmeas, de diferentes idades foram estudadas. Somente olhos hígidos foram utilizados. Aleatoriamente, um olho foi incluído no grupo controle e outro no grupo tratamento. Os olhos do grupo tratamento foram expostos durante um minuto à indocianina verde a 0,5%. Posteriormente, o endotélio da córnea foi corado com azul de tripano e vermelho de alizarina, analisado e fotografado usando microscópio óptico. As áreas com células endoteliais danificadas foram aferidas e quantificadas utilizando um software para análise morfométrica. Os valores encontrados foram expressos como percentual de perda celular. Em todos os olhos examinados foram observadas áreas de dano celular, tanto no grupo controle quanto no grupo tratamento. A perda celular endotelial média foi de 0,8±0,37% no grupo tratamento e 0,97 ± 0,39% no grupo controle. O teste Qui-quadrado confirmou que os grupos tratamento e controle não diferiram. Foi possível concluir que a ICG 0,5% não causou dano agudo nas células do endotélio da córnea de equinos. Palavras-chave: Catarata. Indocianina verde. Endotélio corneano. Vermelho de alizarina.

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An Easy and Inexpensive Method for Quantitative Analysis of Endothelial Damage by Using Vital Dye Staining and Adobe Photoshop Software

Cited by 68 publications

References 20 publications

Morphology of endothelial cells from different regions of the equine cornea

Morphology of endothelial cells from different regions of the equine cornea

Carbonic Anhydrase IX is Not a Predictor of Outcomes in Non-Metastatic Clear Cell Renal Cell Carcinoma - A Digital Analysis of Tissue Microarray

Evaluation of equine corneal endothelium after exposure to 0.5% indocyanine green - in vitro study

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