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Time-lapse Microscopy - An Introduction


The computerised automation of microscopes and cameras, as well as other devices like stages, filters and shutters, means that experiments can be carried out with live specimens on microscopes for long periods of time without the operator needing to be present. The automation capabilities of microscope systems can vary from the basic ability to capture a sequence of images using a camera, to synchronised control of high speed devices like Yokogawa discs, piezo focus devices and lasers. Almost all microscopes at the LMCB are capable of the most basic time-lapse acquisition but experiments involving a high degree of automation must be carried out using specialised microscope systems and the precise nature of the time-lapse will determine the machine or machines of choice.

Since the specimen is usually alive there is also the need to control the environment around the microscope stage so the specimen remains viable during the time-lapse. This is especially critical for long time-lapses of a day or more. Microscope incubation systems have been developed both by research groups and by several companies to maintain the environment at a constant temperature. The usual requirement is for 37°C for mammalian cells, which will often also be cultured in bicarbonate containing buffer requiring 5% or 10% CO2 , so a means of delivering gas at these concentrations is also required. The LMCB has dedicated time-lapse microscopes with environmental control that can be used for long-term imaging of cells using air lenses or high resolution immersion lenses and specimens in a variety of culture vessels. Multi-well plates are the most popular but 35 mm dishes with glass bottoms can also be accommodated, as can Lab-Tek chambered coverslips or ibidi µSlides.



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