A temperature-controlled stage for laser scanning confocal microscopy and case studies in materials science

If confocal microscopy is an ubiquitous tool in life science, its applications in chemistry and materials science are still, in comparison, very limited. Of particular interest in these domains is the use of confocal microscopy to investigate temperature-dependent phenomena such as self-assembly, diffusio- or thermophoresis, or crystal growth. Several hurdles must be solved to develop a temperature-controlled stage for laser scanning confocal microscopy, in particular regarding the influence of an elevated temperature gradient close to the microscope objective, which most people try very hard to avoid. Here we report the design of a temperature-controlled stage able to generate stable temperature gradients in both positive and negative temperature range and does not require use of liquid nitrogen. Our setup provides an excellent control of the temperature gradient, which can be coupled with a controlled displacement of the sample, making it useful in particular for a variety of solidification, chemistry, and interfacial problems. We illustrate the benefits of our setup with several case studies of interest in chemistry and materials science: the 3D real-time imaging of ice growth, the segregation of hard particles by growing crystals, the freezing behaviour of single emulsions, the self-shaping of oil droplets upon cooling, and the self-assembly of amphiphile molecules into helical structures. These results show how confocal microscopy coupled with a temperature-controlled stage that provides a controlled temperature gradient is a welcome addition to the toolkit of chemists and materials scientists.