Specimen Contrast in Microscopy

Examine the origins of contrast in a wide spectrum of specimens.

Phase Contrast Microscopy

Using refractive index variations to produce optical contrast in transparent specimens.

Fluorescence Microscopy

Examining specimens labeled with molecules that absorb light and emit fluorescence.

Autofluorescence

Natural fluorescence in plants and animals that can also be introduced by fixatives.

Polarized Light Microscopy

Observing birefringent specimens using cross-polarized illumination.

Microscope Optical Systems

Information on the basic properties of microscope and camera optics.

Differential Interference Contrast (DIC)

Contrast enhancement using interference of polarized light wavefronts.

Confocal Microscopy

Imaging with laser excitation and pinhole detection of fluorescence.

Spinning Disk Microscopy

Excellent technique for high-speed imaging of living cells in real time with a CCD camera.

Stereomicroscopy

Long working distance, low magnification microscopes for stereoscopic observation.

General Phase Contrast Literature Sources

A listing of pertinent books, review articles, and research reports on phase contrast.

Linear Measurements (Micrometry)

Periodical location information about micrometry and stereology, as well as a listing of selected review articles and books describing quantitative microscopy.

Resonant Scanning Confocal Microscopy

Resonant scanning confocals can gather images at 30 frames per second or higher.

Multiphoton Microscopy

Unique excitation scheme that reduces photobleaching and phototoxicity.

Spectral Imaging and Linear Unmixing

A powerful technique for co-localization, FRET, and removing autofluorescence.

Fluorophore Co-Localization

Detecting the presence of two or more molecules at the same physical location.

Deconvolution Microscopy

A powerful tool for three-dimensional analysis of complex biological specimens.

Fluorescence Resonance Energy Transfer

Probing molecular interactions in living cells using fluorescent proteins and fluorophores.

Total Internal Reflection Microscopy

TIRFM is useful for probing cellular processes that occur near the membrane.

Single Molecule Fluorescence Microscopy

This technique is emerging as a powerful tool for superresolution imaging.

Fluorescent Speckle Microscopy

Stochastic but sparse labeling of macromolecules to track protein dynamics.

FLIM Literature References

Imaging to resolve fluorescence lifetimes with high spatial resolution.

Superresolution Techniques

Breaking the diffraction barrier to image molecules with nanometer precision.

Fluorescence Resonance Energy Transfer

Probing molecular interactions in living cells using fluorescent proteins and fluorophores.

Fluorescence Recovery after Photobleaching

FRAP is used to determine the mobility of fluorescently tagged proteins in live cells.

Live-Cell Imaging

Basic concepts in maintaining and imaging living cells on the microscope stage.

Focus Drift in Optical Microscopy

Focus drift describes the inability to maintain focus over an extended period of time.

Cellular Phototoxicity

Live-Cell Imaging Chambers

Chambers specially designed to keep cells alive and healthy on the microscope stage.

FRET Microscopy with Spectral Imaging

Excellent for ratiometric analysis of photobleaching and spectral imaging FRET.

Electron-Multiplying CCD Cameras

EMCCDs are the detector system of choice for live-cell imaging in real-time applications.

Fluorescent Proteins

Fluorescent proteins are currently the probes of choice for live-cell imaging.

Synthetic Fluorophores and Hybrid Tags

A number of fluorophores have been developed for imaging living and fixed cells.

Fluorescent Protein Biosensors

Genetically-encoded FRET biosensors are useful in detecting a many biological events.

Quantum Dots

Nanocrystals with excellent spectral properties and extreme photostability.

Optical Highlighter Fluorescent Proteins

Genetically-encoded probes capable of activation, switching, and photoconversion.

Fluorophore Photobleaching

Understanding the mechanisms and kinetics of fluorescence decay in fluorescent probes.

Bimolecular Fluorescence Complementation

Detecting molecular interactions in live cells with split fluorescent proteins.

Chromophore-Assisted Light Inactivation

CALI is a useful technique for selectively deactivating target molecules.

Superresolution Review Articles

Overviews of the literature written by experts in optical design and superresolution imaging.

3D Superresolution Techniques

Probing both the lateral and axial dimensions at resolutions beneath the diffraction limit.

dSTORM Microscopy

A single-molecule superresolution technique that utilizes conventional fluorophores.

STORM Microscopy

Single-molecule superresolution using photoswitchable carbocyanine dyes.

Stimulated Emission Depletion (STED)

A point-spread function engineering technique that relies on high-power lasers.

Superresolution Structured Illumination

Applying structured excitation illumination to resolve high spatial frequencies.

Interference Superresolution Microscopy

Saturated Structured Illumination

Structured illumination excitation with theoretically infinite resolution.

Superresolution Practical Aspects

Points to consider when conducting single-molecule superresolution investigations.

PALMIRA Superresolution Imaging

An advanced single-molecule technique that avoids several potential artifacts.

Structured Illumination Microscopy

Patterned excitation for optical sectioning forms the basis for superresolution.

Selective Plane Illumination

An emerging technique with great potential for superresolution imaging.

The RESOLFT Concept

Describing superresolution with reversible saturable or switchable optical transitions.

4Pi Superresolution Microscopy

Using opposed objectives to narrow the axial point spread function to near 100 nanometers.

Ground State Depletion (GSD) Microscopy

GSD relies on driving excited state molecules into a dark metastable long-lived triplet state.

PALM Microscopy

Single-molecule superresolution using optical highlighter fluorescent proteins.

Standing Wave Microscopy

Axial resolution enhancement using interference between two standing waves.

Single-Molecule Localization

Fundamental principles underpinning the techniques of PALM, STORM, and GSDIM.

Live-Cell Superresolution

Investigations are beginning to address dynamics using superresolution microscopy.

Superresolution Microscopy Probes

Fluorescent proteins, synthetics, quantum dots, and hybrid systems for superresolution.

Superresolution Localization Accuracy

Critical aspects of stage drift, molecular density, background signal and other artifacts.

Blinking Superresolution Microscopy

Single-molecule imaging tuned to the fluctuating emission of fluorophores.

Specialized Superresolution Methods

Emerging methodology is now being examined for potential applications.

Near-Field Scanning (NSOM)

Probing specimens with an evanescent wavefield for superresolution.