Confocal microscope

Beskrivning

Intention to purchase a Zeiss LSM 980 with airyscan, joint deconvolution, dynamics profiling and temperature control. The department of Forest Genetics and Plant Physiology (Umeå Plant Science Centre) intends to purchase a Super-Resolution Confocal Microscope, coupled with a temperature-controlled stage/incubation system and advanced spectroscopic techniques to study membrane fluidity, dynamics and molecular aggregates under temperature stress in live whole seedling specimens in deeper layer of Arabidopsis thaliana with enough imaging speed to be able to capture the events. The main reason for this choice of system is the feature of dynamics profiling. Previously fluorescence correlation spectroscopy (FCS) was used to measure flow in one single excitation volume, but this method has limitations in that it cannot provide spatial information. With dynamics profiler that uses a 32-channel detector one can collect 32 individual FCS intensity traces at once in a spatial area. This makes it possible to study asymmetric or directional diffusion data and provide flow data of proteins within the cells for directional information. This is something we find very useful for our main research focus for this instrument on deciphering the organization and fluidity of membranes where dynamics profiling will enable us to precisely analyze the heterogeneous motion of fluorescently labeled proteins, gain insights into molecular aggregations in the membranes, and measure dynamic co-localization and molecular interactions. Further the system needs to provide superb image quality and resolution, while ensuring the preservation of our valuable samples to allow live imaging of whole plants. 32-channel airyscan with joint deconvolution provides high resolution, can be used with single fluorophores, and works with the difficult live plant samples. The experiments need to be conducted under highly controlled environmental conditions. Therefore, the confocal microscope should be outfitted with a temperature-controlled incubation system (from 10°C to 40°C) and imaging software that grants us the capability to meticulously manage and document all experimental conditions, enabling us to correlate results with specific conditions effectively. In our pursuit of investigating dynamic processes, we need a super-resolution confocal microscope capable of faster imaging compared to standard confocal microscopy. The 8-line parallel multiplex mode of the Zeiss LSM 980 provide us with enough speed for this. Based on these factors of resolution, speed, environmental control and mainly dynamics spectroscopy, SLU intend to purchase a Zeiss LSM 980 system with airyscan 2, multiplex SR-8Y, dynamics profiler and temperature control.