Photobleaching imprinting microscopy: seeing clearer and deeper.
نویسندگان
چکیده
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 twofold 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. Moreover, we show that PIM can increase the image contrast of biological tissue, effectively extending the fundamental depth limit of multi-photon fluorescence microscopy.
منابع مشابه
Optical sectioning by wide-field photobleaching imprinting microscopy.
We present a generic wide-field optical sectioning scheme, photobleaching imprinting microscopy (PIM), for depth-resolved cross-sectional fluorescence imaging. Wide-field PIM works by extracting a nonlinear component that depends on the excitation fluence as a result of photobleaching-induced fluorescence decay. Since no specific fluorescent dyes or illumination modules are required, wide-field...
متن کاملNonlinear light-sheet fluorescence microscopy by photobleaching imprinting.
We present a nonlinear light-sheet fluorescence microscopy (LSFM) scheme based on photobleaching imprinting. By measuring photobleaching-induced fluorescence decay, our method simultaneously achieves a large imaging field of view and a thin optical section. Furthermore, the scattered-light-induced background is significantly reduced, considerably improving image contrast. Our method is expected...
متن کاملFluorescence Microscopy Gets Faster and Clearer: Roles of Photochemistry and Selective Illumination
Significant advances in fluorescence microscopy tend be a balance between two competing qualities wherein improvements in resolution and low light detection are typically accompanied by losses in acquisition rate and signal-to-noise, respectively. These trade-offs are becoming less of a barrier to biomedical research as recent advances in optoelectronic microscopy and developments in fluorophor...
متن کاملInducible fluorescent speckle microscopy
The understanding of cytoskeleton dynamics has benefited from the capacity to generate fluorescent fiducial marks on cytoskeleton components. Here we show that light-induced imprinting of three-dimensional (3D) fluorescent speckles significantly improves speckle signal and contrast relative to classic (random) fluorescent speckle microscopy. We predict theoretically that speckle imprinting usin...
متن کاملReal Time 3D fluorescence microscopy based on multifocal multiphoton excitation
Multiphoton excitation microscopy is becoming increasingly important as it offers several advantages over confocal laser (single photon excitation) scanning microscopy (CLSM) such as use of lesser damaging NIR wavelengths, reduced scattering, deeper imaging, no need for pinhole, higher light collection efficiency and excitation of UV dyes at NIR wavelengths; in addition, excitation and photoble...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Journal of cell science
دوره 127 Pt 2 شماره
صفحات -
تاریخ انتشار 2014