Nonlinear light-sheet fluorescence microscopy by photobleaching imprinting

We present a generic sub-diffraction-limited imaging method -photobleaching imprinting microscopy (PIM) -for biological fluorescence imaging. A lateral resolution of 110 nm was measured, more than a two-fold improvement over the optical diffraction limit. Unlike other super-resolution imaging techniques, PIM does not require complicated illumination modules or specific fluorescent dyes. PIM is expected to facilitate the conversion of super-resolution imaging into a routine lab tool, making it accessible to a much broader biological research community.

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“…sub-diffractio llular imaging per-resolution yes, making su convolution m ratio [14]. M can be pot SFM) [16] …”

Section: Resultsmentioning

confidence: 99%

Sub-diffraction-limited imaging by photobleaching imprinting microscopy

Gao

Garcia-Uribe

Liu³

et al. 2014

SPIE Proceedings

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“…Zebrafish husbandry was described in our previous work [21]. All animal work was performed in compliance with Washington University’s institutional animal protocols.…”

Section: Resultsmentioning

confidence: 99%

Multiview optical resolution photoacoustic microscopy

Zhu¹,

Li²,

Gao³

et al. 2014

Optica

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Optical resolution photoacoustic microscopy (OR-PAM), while providing high lateral resolution, has been limited by its relatively poor acoustically determined axial resolution. Although this limitation has been tackled in recent works by using either broadband acoustic detection or nonlinear photoacoustic effects, a flexible solution with three dimensional optical resolution in reflection mode remains desired. Herein we present a multi-view OR-PAM technique. By imaging the sample from multiple view angles and reconstructing the data using a multi-view deconvolution method, we have experimentally demonstrated an isotropic optical resolution in three dimensions.

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“…Fluorophores irreversibly lose their ability to fluorescence due to the photon-induced chemical damage and covalent modification. The photobleaching obeys an empirical exponential temporal decay law (Gao et al, 2014b) (shown in Figure 2A) and in the Nth frame captured at time t n , the absorption rate µ is also a function of t n , written as:…”

Section: Photobleaching Imprinting Of Ltfmmentioning

confidence: 99%

Photobleaching Imprinting Enhanced Background Rejection in Line-Scanning Temporal Focusing Microscopy

Zhuang

Zhang

et al. 2020

Front. Chem.

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Compared with two-photon point-scanning microscopy, two-photon temporal focusing microscopy (2pTFM) provides a parallel high-speed imaging strategy with optical sectioning capability. Owing to out-of-focus fluorescence induced by scattering, 2pTFM suffers deteriorated signal-to-background ratio (SBR) for deep imaging in turbid tissue, Here, we utilized the photobleaching property of fluorophore to eliminate out-of-focus fluorescence. According to different decay rates in different focal depth, we extract the in-focus signals out of backgrounds through time-lapse images. We analyzed the theoretical foundations of photobleaching imprinting of the line-scanning temporal focusing microscopy, simulated implementation for background rejection, and demonstrated the contrast enhancement in MCF-10A human mammary epithelial cells and cleared Thy1-YFP mouse brains. More than 50% of total background light rejection was achieved, providing higher SBR images of the MCF-10A samples and mouse brains. The photobleaching imprinting method can be easily adapted to other fluorescence dyes or proteins, which may have application in studies involving relatively large and nontransparent organisms.

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Nonlinear light-sheet fluorescence microscopy by photobleaching imprinting

Cited by 12 publications

References 15 publications

Sub-diffraction-limited imaging by photobleaching imprinting microscopy

Sub-diffraction-limited imaging by photobleaching imprinting microscopy

Multiview optical resolution photoacoustic microscopy

Photobleaching Imprinting Enhanced Background Rejection in Line-Scanning Temporal Focusing Microscopy

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