Abstract
Macro photography is applicable for imaging various tissue samples at high magnification to perform qualitative and quantitative analyses. Tissue preparation and subsequent image capture are steps performed immediately after the ischemia-reperfusion (IR) experiment and must be performed in a timely manner and with appropriate care. For the evaluation of IR-induced damage in the heart and brain, this paper describes 2,3,5-triphenyl-2H-tetrazolium chloride (TTC)-based staining followed by macro photography. Scientific macro photography requires controlled lighting and an appropriate imaging setup. The standardized methodology ensures high-quality, detailed digital images even if a combination of an inexpensive up-to-date digital camera and macro lens is used. Proper techniques and potential mistakes in sample preparation and image acquisition are discussed, and examples of the influence of correct and incorrect setups on image quality are provided. Specific tips are provided on how to avoid common mistakes, such as overstaining, improper sample storage, and suboptimal lighting conditions. This paper shows the appropriate methodology for rat heart and brain tissue slicing and staining and provides guidelines for establishing lighting and camera setups and photography techniques for high-resolution image acquisition.
Highlights
Photography and analysis of heart and brain tissue specimens have been an important part of life science experiments
Freezing for a prolonged period (>1 h, Figure 3C) reduces mitochondrial function; tetrazolium chloride (TTC) staining of the heart is not red but pale pink, and the border between necrotic and viable tissues is blurred (Figure 3C)
Preparation of the heart after IR starts with the reocclusion of blood cardiac arteries and the perfusion of blue dye for the discrimination of at-risk areas from non-risk areas
Summary
Photography and analysis of heart and brain tissue specimens have been an important part of life science experiments. Science and innovation progress drives the development of expensive microscopes capable of superresolution. Photomicrographs are obtained in a wellcontrolled light environment following detailed instructions. Macro photography (at 1:2 or greater magnification) is frequently performed in an uncontrolled light environment using inappropriate imaging setups. The techniques of sample preparation and camera setup need to be substantially optimized. Macro photographs of limited quality have been widely published in scientific
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