Скачать или смотреть Collect and fix adult Drosophila eyes using appropriate fixatives to preserve cellular structures

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The process of preparing adult Drosophila eyes for thin sectioning and subsequent microscopic analysis is a meticulous procedure that requires careful attention to detail. Initially, the adult flies must be collected and anesthetized, typically using carbon dioxide to ensure minimal stress and damage to the specimens. Once anesthetized, the eyes are carefully excised from the head using fine dissection tools, ensuring that the surrounding tissues are preserved as much as possible. This step is crucial, as the integrity of the eye structure is essential for accurate microscopic examination. The excised eyes are then placed in a fixative solution, such as 4% paraformaldehyde, to preserve cellular morphology and prevent degradation during the subsequent processing steps.

Following fixation, the eyes undergo a series of dehydration and embedding steps to prepare them for thin sectioning. The fixation solution is replaced with a series of graded ethanol solutions, gradually increasing in concentration to facilitate the removal of water from the tissues. After dehydration, the eyes are infiltrated with a suitable embedding medium, such as paraffin wax or resin, which provides structural support during sectioning. This infiltration process may take several hours to ensure complete penetration of the embedding medium into the eye tissues. Once adequately embedded, the samples are cooled and solidified, allowing for the precise cutting of thin sections using a microtome.

The final stage involves sectioning the embedded eyes into ultra-thin slices, typically ranging from 5 to 10 micrometers in thickness. These sections are then mounted onto glass slides for microscopic analysis. It is essential to ensure that the sections are cut evenly to maintain the integrity of the cellular structures for observation. After mounting, the sections may be stained with specific dyes to enhance contrast and highlight particular cellular components, facilitating detailed examination under a microscope. This comprehensive preparation process is vital for obtaining high-quality images and data, enabling researchers to study the intricate structures and functions of Drosophila eyes effectively.