For correlative imaging, here combining cryogenic fluorescence microscopy (cryo-FM) and cryogenic transmission electron microscopy (cryo-TEM), we developed the following workflow (Fig. ). First, we prepared the vitrified samples described in section 2.3 (Fig. ). Then, we transferred the vitrified specimen to the LINKAM cryo-stage and obtained cryo-FM images (Fig. ). The sample loading step is conducted in the LINKAM cryostage, from a grid box to the loading station, then to the sample stage. The EM grid is inserted into the sample magnet clamp in the loading station, which is then loaded on the sample stage. The liquid nitrogen (LN2) reservoir is connected to the stage, keeping the temperature of the sample stage at 77 K. A list of the optical components for cryo-FM is given in section 2.4, and the optical setup will be elucidated in the next section. After the measurements in cryo-FM, we transferred the sample to cryo-TEM. Cryo-TEM is conducted on the same regions identified by cryo-FM. We used the holey carbon film to increase the contrast, and a brief explanation of cryo-TEM is provided in Fig. . Accelerated electron beams generally travel through three sets of electromagnetic lenses (condenser, objective, and projection lenses) before the magnified image of a specimen is obtained .s were first diluted and vitrified using plunge vitrification. Prior to vitrification, the TEM grid is first plasma treated to make the carbon surface hydrophilic. Then the diluted sample was applied, blotted with filter paper, and plunged into liquid ethane. These steps were conducted using a Vitrobot Mark IV (section 2.3). The vitrified samples were stored in a grid box at 77 K in liquid nitrogen before cryo-FM measurements. (b) The cryo-sample is loaded and kept at cryogenic temperatures using a LINKAM CMS196 cryo stage during cryo-FM measurements. After fluorescence imaging at 77 K, sample positions are mapped to retrieve the same position in cryo-TEM later. (c) The cryo-sample is loaded in cryo-TEM, and high-resolution images of the targeted area are obtained. To achieve higher TEM imaging contrast, holey carbon films were used.