Cryo-SOFI enabling low-dose super-resolution correlative light and electron cryo-microscopy
Map data
Tomogram of pseudopod of an XC cell. This cell was imaged with cryo-fluorescence microscopy super-resolution optical fluctiation imaging (cryoSOFI) prior to tilt-series acquisition.
Journal: Proc Natl Acad Sci U S A / Year: 2019 Title: Cryo-SOFI enabling low-dose super-resolution correlative light and electron cryo-microscopy. Authors: Felipe Moser / Vojtěch Pražák / Valerie Mordhorst / Débora M Andrade / Lindsay A Baker / Christoph Hagen / Kay Grünewald / Rainer Kaufmann / Abstract: Correlative light and electron cryo-microscopy (cryo-CLEM) combines information from the specific labeling of fluorescence cryo-microscopy (cryo-FM) with the high resolution in environmental context ...Correlative light and electron cryo-microscopy (cryo-CLEM) combines information from the specific labeling of fluorescence cryo-microscopy (cryo-FM) with the high resolution in environmental context of electron cryo-microscopy (cryo-EM). Exploiting super-resolution methods for cryo-FM is advantageous, as it enables the identification of rare events within the environmental background of cryo-EM at a sensitivity and resolution beyond that of conventional methods. However, due to the need for relatively high laser intensities, current super-resolution cryo-CLEM methods require cryo-protectants or support films which can severely reduce image quality in cryo-EM and are not compatible with many samples, such as mammalian cells. Here, we introduce cryogenic super-resolution optical fluctuation imaging (cryo-SOFI), a low-dose super-resolution imaging scheme based on the SOFI principle. As cryo-SOFI does not require special sample preparation, it is fully compatible with conventional cryo-EM specimens, and importantly, it does not affect the quality of cryo-EM imaging. By applying cryo-SOFI to a variety of biological application examples, we demonstrate resolutions up to ∼135 nm, an improvement of up to three times compared with conventional cryo-FM, while maintaining the specimen in a vitrified state for subsequent cryo-EM. Cryo-SOFI presents a general solution to the problem of specimen devitrification in super-resolution cryo-CLEM. It does not require a complex optical setup and can easily be implemented in any existing cryo-FM system.
Download / File: emd_4471.map.gz / Format: CCP4 / Size: 362.4 MB / Type: IMAGE STORED AS SIGNED INTEGER (2 BYTES)
Annotation
Tomogram of pseudopod of an XC cell. This cell was imaged with cryo-fluorescence microscopy super-resolution optical fluctiation imaging (cryoSOFI) prior to tilt-series acquisition.
Voxel size
X=Y=Z: 16.88 Å
Density
Minimum - Maximum
-32767.0 - 32767.0
Average (Standard dev.)
14363.378000000000611 (±1587.113499999999931)
Symmetry
Space group: 1
Details
EMDB XML:
Map geometry
Axis order
X
Y
Z
Origin
-39
-62
110
Dimensions
849
835
268
Spacing
835
849
268
Cell
A: 14094.799 Å / B: 14331.119 Å / C: 4523.84 Å α=β=γ: 90.0 °
Material: GOLD / Mesh: 200 / Support film - #0 - Film type ID: 1 / Support film - #0 - Material: CARBON / Support film - #0 - topology: LACEY / Support film - #1 - Film type ID: 2 / Support film - #1 - Material: GRAPHENE OXIDE / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Atmosphere: AIR
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