Abstract
This article presents the study and recommendation of a method to improve the observation, in laboratory experiments, using a microscope stereoscopic or stereomicroscope with chicken embryo chorioallantoic membranes (CAM). In these experiments, tumor cells are implanted in this CAM so that they will form a tumor during the embryo's development period. The number of neovessels that develop around the tumors are indicators of results on testing with antiangiogenic drugs. These studies require quantification of vascularization to compare the stimulating or inhibitory effects caused by different agents. The proposed method to improve the visualization of the vessels consists of a lighting chamber composed of three lighting rings, to accommodate the egg, for observation in a microscope. Each ring is composed of sets of 8–16 LEDs, with a temperature of color between 3000 and 5500 K, aligned, focusing on the regions above, central and below the egg. The method provided an improvement in the results as more details of the images were observed, when compared to the images obtained with the original illumination of the adopted microscope stereoscopic or stereomicroscope, model LEICA ZOOM™ 2000. Also, the proposed LED lighting avoids the heating irradiated by the original illumination, which may cause discomfort, dehydration and unnecessary movements of the embryo, besides reflection or glare on the exposed membranes and liquids of the egg, which may impairment the visualization. Qualitative evaluation of sample images demonstrated that subtle vessels stand out, thus allowing better results for further quantitative analysis of the CAM. The quantitative analysis of the images, using the vessel counting method based on a grid, resulted in a mean percentage difference counting of +56% vessels in relation to the counting with the original illumination of the microscope.
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References
Jain RK, Schlenger K, Hockel M et al (1997) Quantitative angiogenesis assays: progress and problems. Nat Med 3:1203–1208. https://doi.org/10.1038/nm1197-1203
Ribatti D, Nico B, Vacca A et al (2006) The gelatin sponge–chorioallantoic membrane assay. Nat Protoc 1:85–92. https://doi.org/10.1038/nprot.2006.13
Doukas CN, Maglogiannis I, Chatziioannou AA (2008) Computer-supported angiogenesis quantification using image analysis and statistical averaging. IEEE Trans Inf Technol Biomed 12:650–657. https://doi.org/10.1109/TITB.2008.926463
Andrade SP, Fan TP, Lewis GP (1987) Quantitative in vivo studies on angiogenesis in a rat sponge model. Br J Exp Pathol 68:755–766
Serbedzija GN, Flynn E, Willet CE (1999) Zebrafish angiogenesis: a new model for drug screening. Angiogenesis 3:353–359. https://doi.org/10.1023/a:1026598300052
Nowak-Sliwinska P, Segura T, Iruela-Arispe ML (2014) The chicken chorioallantoic membrane model in biology, medicine and bioengineering. Angiogenesis 17:779–804. https://doi.org/10.1007/s10456-014-9440-7
Bo Z, Liya A, Shao L et al (2009) AngioIQ: a novel automated analysis approach for angiogenesis image quantification. In: 2nd international conference on biomedical engineering and informatics, Tianjin, 2009, pp 1–5. https://doi.org/10.1109/BMEI.2009.5304911
Ribatti D, Tamma R (2018) The chick embryo chorioallantoic membrane as an in vivo experimental model to study human neuroblastoma. J Cell Physiol 2018:1–6. https://doi.org/10.1002/jcp.26773
Leica Microsystems at https://www.leica-microsystems.com
Sales RP (2011) LED, o novo paradigma da iluminação pública, LACTEC/IEP
Oliveira CHL, Costa MAD, Costa GH (2015) Comparação entre a Lâmpada Halógena e o LED como fontes de Iluminação na Microscopia Óptica, 8o Congresso Brasileiro de Metrologia. Bento Gonçalves 2015:1–4
Kerr PH, Fisher EM, Buffington ML (2008) Dome lighting for insect imaging under a microscope. Am Entomol 54:198–200. https://doi.org/10.1093/ae/54.4.198
UNICAMP at https://tinyurl.com/y6xydxj9
Waide P (2010) Phase out of incandescent lamps Implications for international supply and demand for regulatory compliant lamps. IEA—International Energy Agency, p 86
Zhang Y, Xie W, Li Z et al (2013) Model predictive direct power control of a PWM rectifier with duty cycle optimization. IEEE Trans Power Electron 28:5343–5351. https://doi.org/10.1109/TPEL.2013.2243846
Jo S-A, Lee C-H, Kim M-J et al (2019) Effect of pulse-width-modulated LED light on the temperature change of composite in tooth cavities. Dent Mater 35:554–563. https://doi.org/10.1016/j.dental.2019.01.009
ARDUINO at https://www.arduino.cc/
Yang S-H, Lin J-K, Huang C-J et al (2005) Silibinin inhibits angiogenesis via Flt-1, but not KDR, receptor up-regulation. J Surg Res 128:140–146. https://doi.org/10.1016/j.jss.2005.04.04
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Silva, M.K., Stoltz, I.R., Rocha, L.T., Pereira, L.F., de Souza, M.A., Borba, G.B. (2022). A Method for Illumination of Chorioallantoic Membrane (CAM) of Chicken Embryo in Microscope. In: Bastos-Filho, T.F., de Oliveira Caldeira, E.M., Frizera-Neto, A. (eds) XXVII Brazilian Congress on Biomedical Engineering. CBEB 2020. IFMBE Proceedings, vol 83. Springer, Cham. https://doi.org/10.1007/978-3-030-70601-2_118
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DOI: https://doi.org/10.1007/978-3-030-70601-2_118
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