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Resolution enhancement for low-temperature scanning microscopy by cryo-immersion

  • Here we report a simple way to enhance the resolution of a confocal scanning microscope under cryogenic conditions. Using a microscope objective (MO) with high numerical aperture (NA = 1:25) and 1-propanol as an immersion fluid with low freezing temperature we were able to reach an imaging resolution at 160 K comparable to ambient conditions. The MO and the sample were both placed inside the inner chamber of the cryostat to reduce distortions induced by temperature gradients. The image quality of our commercially available MO was further enhanced by scanning the sample (sample scanning) in contrast to beam scanning. The ease of the whole procedure marks an essential step towards the development of cryo high-resolution microscopy and correlative light and electron cryo microscopy (cryoCLEM).

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Metadaten
Author of HS ReutlingenBrecht, Marc
URN:urn:nbn:de:bsz:rt2-opus4-10282
DOI:https://doi.org/10.1364/OE.24.013023
ISSN:1094-4087
Erschienen in:Optics express
Publisher:Optical Society of America
Place of publication:Washington, DC
Document Type:Article
Language:English
Year of Publication:2016
Tag:confocal microscopy; fluorescence microscopy; scanning microscopy; spatial resolution
Volume:24
Issue:12
Page Number:10
First Page:13023
Last Page:13032
DDC classes:530 Physik
Open Access?:Ja
Licence (German):License Logo  Open Access