Abstract
The goal of biological samples’ cryofixation is to trap a metabolic state as it exists in vivo by rapidly stopping internal reactions. However, obtaining perfect quality of cryofixation for large and high hypermetabolism organ/tissue (such as brain, heart) remains a challenge. The aim of this study was to develop and display a comprehensive and direct method to evaluate cryofixation’s process and quality. Here, we adopt a delicate combination of homemade cryo-imaging system with a rat cardiac arrest model that can control cryofixation time optionally. we successfully evaluate the cryofixation time-related nicotinamide adenine dinucleotide (NADH) fluorescence pattern of several coronal sections in rat’s brain that suffered from directional funnel cryofixation procedure. Through quantitative analysis of the distribution map of NADH fluorescence, we could obtain a relationship between cryofixation time and well cryofixation volume and then could deduce the cryofixation rates and quality at different time points. Our results also demonstrated that dissection of the temporal muscle of rat could significantly optimize the classical direct funnel cryofixation protocol.
Nannan Sun and Weihua Luo have contributed equally.
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Acknowledgments
This work was supported by the National Major Scientific Research Program of China (Grant No. 2011CB910401) and the Science Fund for Creative Research Group of China (Grant No.61121004) and the Director Fund of Wuhan National Laboratory for Optoelectronics and the Specific International Scientific Cooperation (Grant No. 2010DFR30820).
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Sun, N., Luo, W., Wang, A., Luo, Q. (2013). Quality Evaluation Method for Rat Brain Cryofixation on the Basis of NADH Fluorescence. In: Van Huffel, S., Naulaers, G., Caicedo, A., Bruley, D.F., Harrison, D.K. (eds) Oxygen Transport to Tissue XXXV. Advances in Experimental Medicine and Biology, vol 789. Springer, New York, NY. https://doi.org/10.1007/978-1-4614-7411-1_58
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DOI: https://doi.org/10.1007/978-1-4614-7411-1_58
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