Cryo-SOFI enabling low-dose super-resolution correlative light and electron cryo-microscopy
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 advantageou...
| Հիմնական հեղինակներ: | , , , , , , , |
|---|---|
| Ձևաչափ: | Journal article |
| Լեզու: | English |
| Հրապարակվել է: |
National Academy of Sciences
2019
|
| _version_ | 1826307273770663936 |
|---|---|
| author | Moser, F Pražák, V Mordhorst, V Andrade, DM Baker, LA Hagen, C Grünewald, K Kaufmann, R |
| author_facet | Moser, F Pražák, V Mordhorst, V Andrade, DM Baker, LA Hagen, C Grünewald, K Kaufmann, R |
| author_sort | Moser, F |
| collection | OXFORD |
| description | 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. |
| first_indexed | 2024-03-07T07:00:27Z |
| format | Journal article |
| id | oxford-uuid:ff902c04-af12-4bba-a96a-0e726a464b10 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T07:00:27Z |
| publishDate | 2019 |
| publisher | National Academy of Sciences |
| record_format | dspace |
| spelling | oxford-uuid:ff902c04-af12-4bba-a96a-0e726a464b102022-03-27T13:45:53ZCryo-SOFI enabling low-dose super-resolution correlative light and electron cryo-microscopyJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:ff902c04-af12-4bba-a96a-0e726a464b10EnglishSymplectic Elements at OxfordNational Academy of Sciences2019Moser, FPražák, VMordhorst, VAndrade, DMBaker, LAHagen, CGrünewald, KKaufmann, RCorrelative 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. |
| spellingShingle | Moser, F Pražák, V Mordhorst, V Andrade, DM Baker, LA Hagen, C Grünewald, K Kaufmann, R Cryo-SOFI enabling low-dose super-resolution correlative light and electron cryo-microscopy |
| title | Cryo-SOFI enabling low-dose super-resolution correlative light and electron cryo-microscopy |
| title_full | Cryo-SOFI enabling low-dose super-resolution correlative light and electron cryo-microscopy |
| title_fullStr | Cryo-SOFI enabling low-dose super-resolution correlative light and electron cryo-microscopy |
| title_full_unstemmed | Cryo-SOFI enabling low-dose super-resolution correlative light and electron cryo-microscopy |
| title_short | Cryo-SOFI enabling low-dose super-resolution correlative light and electron cryo-microscopy |
| title_sort | cryo sofi enabling low dose super resolution correlative light and electron cryo microscopy |
| work_keys_str_mv | AT moserf cryosofienablinglowdosesuperresolutioncorrelativelightandelectroncryomicroscopy AT prazakv cryosofienablinglowdosesuperresolutioncorrelativelightandelectroncryomicroscopy AT mordhorstv cryosofienablinglowdosesuperresolutioncorrelativelightandelectroncryomicroscopy AT andradedm cryosofienablinglowdosesuperresolutioncorrelativelightandelectroncryomicroscopy AT bakerla cryosofienablinglowdosesuperresolutioncorrelativelightandelectroncryomicroscopy AT hagenc cryosofienablinglowdosesuperresolutioncorrelativelightandelectroncryomicroscopy AT grunewaldk cryosofienablinglowdosesuperresolutioncorrelativelightandelectroncryomicroscopy AT kaufmannr cryosofienablinglowdosesuperresolutioncorrelativelightandelectroncryomicroscopy |