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Posterior approach to correct for focal plane offsets in lattice light-sheet structured illumination microscopy.
Shi, Yu; Daugird, Tim A; Legant, Wesley R.
Afiliación
  • Shi Y; University of North Carolina at Chapel Hill and North Carolina State University, Joint Department of Biomedical Engineering, Chapel Hill, North Carolina, United States.
  • Daugird TA; University of North Carolina at Chapel Hill, Department of Pharmacology, Chapel Hill, North Carolina, United States.
  • Legant WR; University of North Carolina at Chapel Hill and North Carolina State University, Joint Department of Biomedical Engineering, Chapel Hill, North Carolina, United States.
J Biomed Opt ; 29(8): 086502, 2024 Aug.
Article en En | MEDLINE | ID: mdl-39086928
ABSTRACT

Significance:

Lattice light-sheet structured illumination microscopy (latticeSIM) has proven highly effective in producing three-dimensional images with super resolution rapidly and with minimal photobleaching. However, due to the use of two separate objectives, sample-induced aberrations can result in an offset between the planes of excitation and detection, causing artifacts in the reconstructed images.

Aim:

We introduce a posterior approach to detect and correct the axial offset between the excitation and detection focal planes in latticeSIM and provide a method to minimize artifacts in the reconstructed images.

Approach:

We utilized the residual phase information within the overlap regions of the laterally shifted structured illumination microscopy information components in frequency space to retrieve the axial offset between the excitation and the detection focal planes in latticeSIM.

Results:

We validated our technique through simulations and experiments, encompassing a range of samples from fluorescent beads to subcellular structures of adherent cells. We also show that using transfer functions with the same axial offset as the one present during data acquisition results in reconstructed images with minimal artifacts and salvages otherwise unusable data.

Conclusion:

We envision that our method will be a valuable addition to restore image quality in latticeSIM datasets even for those acquired under non-ideal experimental conditions.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Imagenología Tridimensional / Microscopía Fluorescente Límite: Animals / Humans Idioma: En Revista: J Biomed Opt Asunto de la revista: ENGENHARIA BIOMEDICA / OFTALMOLOGIA Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Imagenología Tridimensional / Microscopía Fluorescente Límite: Animals / Humans Idioma: En Revista: J Biomed Opt Asunto de la revista: ENGENHARIA BIOMEDICA / OFTALMOLOGIA Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos