What does two-photon microscopy do?

Two-photon microscopy (also called multiphoton microscopy) can be used for live cell imaging of thick biological specimens, as it has several advantages over confocal microscopy. Molecules can be visualized deeply within the specimen with a maximal penetration depth of about 1 mm.

How much does a two-photon cost?

A base model, turnkey two-photon microscope will cost an additional $125,000 on top of the laser. “By the time you add components, you’re starting at around $225,000 [for the microscope],” says Rafter.

What is the resolution of two-photon microscopy?

STED microscopy has been done with two-photon excitation and has achieved a spatial resolution of ~60 nm in optical systems with two lasers [13–15] and a single laser [16,17]. However, the imaging depth of two-photon STED microscopy is still limited to being less than ~100 µm.

How does two-photon excitation microscopy work?

The concept of two-photon excitation is based on the idea that two photons, of comparably lower photon energy than needed for one photon excitation, can also excite a fluorophore in one quantum event. Each photon carries approximately half the energy necessary to excite the molecule.

What is a 2 photon laser?

In Two-Photon Excitation (TPE), a high power pulsed laser with very short pulse width is focused into the sample. The high photon density in the focus leads to a certain probability that a fluorophore absorbs two photons quasi simultaneously.

Is two-photon microscope a light microscope?

Two-photon microscopy, also referred to as two-photon laser scanning microscopy, is a further refinement of precision fluorescence microscopy. In epifluorescent or confocal microscopy, a single photon of light at a specific wavelength excites a fluorophore to emit light.

What is the disadvantage of confocal microscopy?

Disadvantages of confocal microscopy are limited primarily to the limited number of excitation wavelengths available with common lasers (referred to as laser lines), which occur over very narrow bands and are expensive to produce in the ultraviolet region.

Why is ZEISS the best?

Sharpness. In addition to superb color reproduction, the clarity and high contrast of a ZEISS lens also helps achieve ultra-sharp imagery. The focus mechanism that ZEISS lenses use also plays a part. ZEISS is unique in that they are one of a few major lens brands that still produces manual focus lenses.

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