High-resolution Microscopy
High-resolution Microscopy - Related Videos
08:31
Three-dimensional Optical-resolution Photoacoustic Microscopy
0 Views •
Cited by 27 •
2011
Optical-resolution photoacoustic microscopy (OR-PAM) is an emerging technology capable of imaging optical absorption contrasts in vivo with cellular resolution and sensitivity. Here, we provide a visualized instruction on the experimental protocols of OR-PAM, including system configuration, system alignment, typical in vivo experimental procedures, and functional imaging schemes.
12:28
Correlative Super-resolution and Electron Microscopy to Resolve Protein Localization in Zebrafish Retina
0 Views •
Cited by 11 •
2017
This protocol describes the necessary steps to obtain subcellular protein localization results on zebrafish retina by correlating super-resolution light microscopy and scanning electron microscopy images.
16:52
Test Samples for Optimizing STORM Super-Resolution Microscopy
0 Views •
Cited by 48 •
2013
We describe the preparation of three test samples and how they can be used to optimize and assess the performance of STORM microscopes. Using these examples we show how to acquire raw data and then process it to acquire super-resolution images in cells of approximately 30-50 nm resolution.
08:53
Expansion Microscopy: High-Resolution Fluorescent Imaging with a Conventional Microscope
0 Views •
Cited by 1 •
2025
Here, we present a detailed protocol combining cryo-fixation and expansion microscopy methods (Cryo-ExM), allowing for high-resolution imaging with structural preservation adapted to various biological samples. Biological samples are frozen, embedded in a swellable polymer, denatured, expanded, and immunolabeled, enabling super-resolution imaging with standard light microscopes.
01:37
Super-resolution Fluorescence Microscopy
0 Views •
2023
Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been developed.
FAQs
Resolution improves when specialized optics and controlled illumination provide more precise information about closely spaced structures than conventional microscopy can capture. Sensitive detectors then record weak or finely localized signals with greater detail. Together, these components help distinguish cellular features that would otherwise appear blended, allowing researchers to relate spatial organization to biological function.
Sensitive detectors capture subtle signals from specimens after the optical system and illumination have shaped the image. Their role is especially important when biological structures or molecular distributions produce limited detectable information. By preserving fine spatial detail, detector performance supports more reliable visualization of organelles, protein patterns, cellular interactions, and changes occurring during biological processes.
Computational image reconstruction analyzes recorded image information to distinguish or localize features that appear closely spaced or partially overlapping. Rather than relying only on the raw optical image, the approach uses processing to improve spatial interpretation. This can clarify molecular distributions and cellular organization, supporting quantitative comparisons between biological conditions when the image data permit reconstruction.
The specimen state should match the biological question and the information required from the image. Living specimens are appropriate when researchers need to examine dynamic processes, whereas preserved specimens can support detailed assessment of structures or distributions in a fixed biological state. The selected optics, illumination, detection, and reconstruction approach must remain compatible with that specimen and objective.
Applications include examining organelles, mapping protein distributions, observing cellular interactions, and following dynamic processes in living specimens. These uses connect spatial patterns with biological activity rather than treating an image as a purely descriptive result. The method therefore supports questions about cell organization, interactions between cells, and how microscopic changes accompany broader biological behavior.
In disease studies, the resulting images can reveal cellular or molecular changes whose spatial organization may relate to altered biological function. In developmental biology, imaging can help track how cellular structures and interactions change as systems develop. Quantitative experiments can then link these microscopic observations to biological outcomes, providing a basis for comparing conditions and testing structure-function relationships.