A cooling assembly for a microscope, comprising: (a) a first circulation system for an objective lens , comprising: a light transparent element ; and a collar comprising a middle portion of the collar , wherein the middle portion of the collar comprises an inner surface capable of sealing engagement with an objective lens and a lower surface capable of sealing engagement with the light transparent element; the collar further comprising at least one gasket between the inner surface of the middle portion of the collar and the objective lens and at least one gasket between the lower surface of the middle portion of the collar and the light transparent element, wherein the objective lens and the light transparent element define a region for circulating immersion fluid ; and the collar further comprising one or more ports for immersion fluid transport in and out of the region; (b) a second circulation system for a sample stage, comprising: an insulated enclosure , comprising: a cold block for sample placement; and one or more ports for transport of a cooling fluid in and out of the insulated enclosure, wherein the cooling fluid has a temperature below -135°C; and (c) coupling fluid for optically coupling the light transparent element with a sample, wherein the coupling fluid has a refractive index of between 1.3 and 1.4 and a thermal conductivity of less than 0.3 W/m°K. (a) a first circulation system for an objective lens , comprising: a light transparent element ; and a collar comprising a middle portion of the collar , wherein the middle portion of the collar comprises an inner surface capable of sealing engagement with an objective lens and a lower surface capable of sealing engagement with the light transparent element; the collar further comprising at least one gasket between the inner surface of the middle portion of the collar and the objective lens and at least one gasket between the lower surface of the middle portion of the collar and the light transparent element, wherein the objective lens and the light transparent element define a region for circulating immersion fluid ; and the collar further comprising one or more ports for immersion fluid transport in and out of the region; comprising: a light transparent element ; and a collar comprising a middle portion of the collar , wherein the middle portion of the collar comprises an inner surface capable of sealing engagement with an objective lens and a lower surface capable of sealing engagement with the light transparent element; the collar further comprising at least one gasket between the inner surface of the middle portion of the collar and the objective lens and at least one gasket between the lower surface of the middle portion of the collar and the light transparent element, wherein the objective lens and the light transparent element define a region for circulating immersion fluid ; and the collar further comprising one or more ports for immersion fluid transport in and out of the region; (b) a second circulation system for a sample stage, comprising: an insulated enclosure , comprising: a cold block for sample placement; and one or more ports for transport of a cooling fluid in and out of the insulated enclosure, wherein the cooling fluid has a temperature below -135°C; and comprising: an insulated enclosure , comprising: a cold block for sample placement; and one or more ports for transport of a cooling fluid in and out of the insulated enclosure, wherein the cooling fluid has a temperature below -135°C; and a cold block for sample placement; and one or more ports for transport of a cooling fluid in and out of the insulated enclosure, wherein the cooling fluid has a temperature below -135°C; and (c) coupling fluid for optically coupling the light transparent element with a sample, wherein the coupling fluid has a refractive index of between 1.3 and 1.4 and a thermal conductivity of less than 0.3 W/m°K. <EOS>