Cryo-workstation for rapid freeze fixation

生物标本的显微成像代表了从分子和细胞生物学到生理学和系统学等各种研究学科的重要组成部分。生物研究所（IRBV）配备了极好的成像设备，包括光学（包括共聚焦）和电子显微镜。为了能够满足用户的需求，该设施需要配备冷冻工作站。许多实验方法都要求固定试样，所得结构越“逼真”，图像质量和数据解释就越好。广泛使用的化学固定方案的主要缺点是引入人工制品，由于固定剂渗透缓慢。低温工作站可以通过接触极冷来快速固定，在几毫秒内将分子固定在空间中。此外，与化学固定相比，抗原性的保存得到了显著增强。在过去的几十年里，微球固定技术的引入导致了显微镜技术的革命性发展。为了跟上技术进步，并为研究人员提供最新的设备，我们要求资金用于冷冻工作站，可以通过快速冷冻或猛击冷冻来快速冷冻固定小型生物标本。它将用于标本制备，包括冷冻替代、冷冻断裂、组织化学和免疫标记。它适用于所有主要类型的显微镜，从光和荧光显微镜到透射电子显微镜和扫描电子显微镜。研究所的几个研究方案将从这种设备的可用性中获益匪浅，这些设备将提供给蒙特里萨大学的研究生和科学家以及外部用户。

Microscopical imaging of biological specimens represents a crucial component of a variety of research disciplines ranging from molecular and cell biology to physiology and systematics. The Institut de recherche en biologie végétale (IRBV) is equipped with an excellent imaging facility that comprises both optical (including confocal) and electron microscopes. To be able to respond to the needs of the users, the facility needs to be complemented with a cryo-workstation. Numerous experimental approaches require the specimen to be fixed, and the more "life-like" the resulting structure is, the better the quality of the image and the interpretation of the data. The widely used chemical fixation protocols have the major drawback of introducing artefacts due to the slow penetration of the fixing agent. A cryo-workstation allows the rapid fixation via contact with extreme cold that fixes the molecules in space within milliseconds. Furthermore, compared to chemical fixation, the preservation of antigenicity is significantly enhanced. The introduction of cyro-fixation has resulted in a revolutionary development in microscopy in the past decades. To keep up with the technical advancement and provide the researchers with an up-to-date facility we request the funds for a cryo-workstation that allows rapid freeze fixation of small biological specimens via plunge or slam freezing. It will be used for specimen preparation involving freeze substitution, freeze fracture, histochemistry, and immunolabel. It is useful for all major types of microscopy ranging from light and fluorescence microscopy to transmission electron and scanning electron microscopy. Several research programs at the IRBV would greatly profit from the availability of this equipment which will be accessible to graduate students and scientists of the Université de Montréal as well as to external users.