DE102021005684A1 - STED method with hollow fiber optic cables - Google Patents

Abstract

A high-resolution fluorescence-based microscopy device and method based on RESOLFT / STED / GSD equipped with flexible, highly flexible (micro) hollow capillaries for beam guidance and shaping is presented in order to reach hard-to-reach places on a sample surface in a polluted measurement environment in a critical atmosphere without the high-energy excitation light (usually and therefore preferably in the UV range) and the fluorescence light (emission light usually and therefore preferably in the Vis/IR range) having high attenuation or dispersion a hollow capillary to and from the hard-to-reach places on the sample surface. The inside of the hollow capillary is provided with a special coating that has been applied to the inside of the hollow capillary by a CVD process, preferably an OMCVD, MOCVD, EOCVD or OECVD process or a CVI process, whereby the guided Excitation and emission light has negligible attenuation and dispersion and also retains its polarization direction. This is a necessary prerequisite to be able to carry out the RESOLFT measurement process.

Description

State of the art:

First, an overview of the currently common high- or super-resolution microscopy methods follows. Many of them are based on fluorescence techniques.

RESOLFT:

According to [1], RESOLFT (Reversible Saturable Optical Fluorescence Transitions) includes the two super-resolution fluorescence microscopy techniques STED and GSD, with STED and GSD representing different realizations of the RESOLFT concept. However, RESOLFT and STED are sometimes equated in the literature or denote different embodiments of the same concept ( DE 10 2011 055 367 A1 , DE 10 2015 105 018 A1 ), see the discussion below.

STED:

The STED (Stimulated Emission Depletion Microscopy) method is based on transitions between energetically different electronic states that take place one after the other [2] - [6].

• - Der unterste oder energetisch niedrigste Zustand Z1:
  • Der energetisch niedrigste Zustand ist ein nicht-fluoreszenter oder nicht-fluoreszenzfähiger Zustand. Er wird auch Dunkelzustand genannt.
• - Der mittlere oder energetisch mittlere Zustand Z2:
  • Der energetisch mittlere Zustand ist ein fluoreszenter oder fluoreszenzfähiger Zustand. Er wird auch Hellzustand genannt.

In order for the STED method to be carried out, it is important that three different (electronic) states or energy levels exist in the sample or in the fluorescent dye marking the sample:

• - The lowest or energetically lowest state Z1:
  • The energetically lowest state is a non-fluorescent or non-fluorescent state. It is also called the dark state.
• - The mean or energetically mean state Z2:
  • The energetically intermediate state is a fluorescent or fluoresceable state. It is also called the light state.

• - Der oberste oder energetisch höchste Zustand Z3:
  • Der energetisch höchste Zustand ist ein fluoreszierender Zustand, da das Atom / Molekül in diesem Zustand ein Fluoreszenzphoton emittieren kann. Auch er ist zusammen mit dem fluoreszenzfähigen oder fluoreszenten Zustand Z2 Teil des Hellzustandes.

The transition from the non-fluorescent state to the fluorescent state is also called turning on, turning on or activation or setting or switching. The transition from the fluorescent state to the non-fluorescent state is also called turning off or turning off or deactivation or switching.

• - The highest or energetically highest state Z3:
  • The energetically highest state is a fluorescent state because the atom/molecule can emit a fluorescence photon in this state. It is also part of the light state together with the fluorescent state Z2.

The transition from the fluorescent state to the fluorescent state is called excitation, the transition from the fluorescent state to the fluorescent state is called de-excitation or stimulation and can occur through stimulated emission of fluorescent light. Radiative transitions are also possible (e.g. quenching).

The activation, excitation, de-excitation and deactivation light can have different wavelengths.

• Als Hellzustand bezeichnet man den fluoreszenzfähigen Zustand Z2 und eventuell auch den fluoreszierenden Zustand Z3, da durch den Übergang zwischen beiden ein FluoreszenzPhoton emittiert wird, welches mittels eines optischen Detektors detektiert werden kann, so dass dadurch ein hochauflösendes Bild entstehen kann. Somit ist der Hellzustand das Gegenteil vom Dunkelzustand.

One additional note to the above definitions:

• The bright state is the fluorescent state Z2 and possibly also the fluorescent state Z3, since the transition between the two emits a fluorescence photon that can be detected by an optical detector, so that a high-resolution image can be created. Thus, the light state is the opposite of the dark state.

In the initial state of the STED method, the fluorescent dye molecules (fluorophores) on the sample surface are in a circular area B1 to be detected in the energetically lowest state Z1 (non-fluorescent state). In a first exposure step, all fluorophores are raised from the non-fluorescent state Z1 to the energetically middle state Z2 (fluorescent state) by incident activation or turn-on light S1 with a corresponding wavelength. In a second exposure step, all fluorophores in the area B1 to be detected are brought from the fluorescent state Z2 to the energetically highest fluorescent state Z3 by incident excitation light S2 with a corresponding wavelength. In a third exposure step, the edge area RB1 of the area B1 is de-excited by stimulated emission with a deactivation light beam S3, which has a zero point in the center and an intensity non-zero at the edges, so that the fluorophores located in this edge area RB1 are separated from the fluorescent state Z3 return to the fluorescent state Z2. This light beam with the characteristic donut profile is also called a STED light beam or also a fluorescence prevention or stimulation light beam.

Since the applied de-excitation light beam S3 has a zero in the center, ie has no intensity in the center, the few or even ideally only a single fluorophore in the fluorescent state Z3 remain there. These can now spontaneously return to the fluorescent state Z2 themselves by fluorescence, ie by emission of a fluorescence photon. However, there is now also the possibility of using a further, additional deactivation beam S4 without a central zero point to selectively de-excite the fluorophores remaining in the central zero point in the fluorescent state Z3 by means of a further stimulated emission, so that one can choose or determine the point in time of the fluorescence emission oneself.

The few fluorophores thus left in the central zero in the fluorescent state Z3 then spontaneously emit or stimulate the fluorescence photons, which then reach the detector surface. Since the emission of fluorescence photons from the fluorophores occurs successively inside and outside the central null, it is thus possible to obtain an image of the surface that has a resolution well below the classical Abbe resolution.

However, from a purely wave-optical point of view, the de-excitation process itself is also diffraction-limited: if the de-excitation were linear with the intensity of the de-excitation beam, no significant improvement or increase in resolution according to Abbe could be achieved [6]. However, the resolution limit can only be overcome by a material effect, namely the saturation of the de-excitation with the de-excitation light beam [6]: as already explained above, the intensity of the de-excitation beam S3 has a central zero point, which is not stepped, but within the cross-section -Beam profile of the depletion beam, the transition from the edge area RB1 with maximum intensity to the center of the centrally located zero point is continuous, so that the intensity is really only equal to zero in a very, very small, centrally located zero point area NB1 of the zero point. However, even the lowest intensities of the depletion beam are sufficient to completely depopulate the fluorescent state Z3, i.e. to convert all excited electrons from the excited state Z3 to the de-excited state Z2. Thus, the fluorophores remain in the fluorescent state Z3, which corresponds to the center of the zero point of the de-excitation light beam, only in a very, very small sub-area, namely in the area of the zero point NB1. The diameter of this sub-area, namely the area of the zero point NB1, is far below the classic diffraction limit. Falling below the classic optical resolution limit according to Abbe is not achieved purely by wave-optical means, but is achieved with the help of an effect inherent to the material, namely saturation.

The depletion beam S3 need not necessarily be circular or rotationally symmetrical with a central null. All other intensity distributions are also conceivable, but they must have at least one zero or at least a local intensity minimum. Several zeros arranged in an array (grid) are also conceivable ( DE 10 2006 009 833 A1 ), other non-point geometric figures or a linear root. The latter would be particularly useful if you have to scan large sample areas and if only one spatial dimension is of interest for a super-resolution display (line scan).

If the substances to be examined are autofluorescent, i.e. they fluoresce on their own without having to add and immobilize a fluorescent dye (fluorophore), then STED and the other processes can also be carried out without a fluorophore, which means that a complex process step (immobilizing the fluorophores) can be omitted.

The optical properties of the fluorescence radiation (wavelength, intensity, half-life, lifetime, polarization, etc.) also depend on the physico-chemical properties of the atomic/molecular environment. In particular, the wavelength of the emitted fluorescence radiation is influenced by the atomic/molecular environment (chemical shift). Thus, conclusions and information about the physico-chemical properties and the atomic/molecular structure of the immediate vicinity of the fluorophore can be drawn from the detected fluorescence radiation, in particular the wavelength, but also from the other optical properties of the emitted fluorescence radiation.

In principle, everything that has been realized in normal or classic fluorescence spectroscopy or microscopy (time-resolved fluorescence measurement, mapping, scanning, lifetime imaging nanoscopy (FLIN) etc.) can also be applied and transferred to STED.

RESOLFT:

In general, RESOLFT is a generic term that includes STED and GSD, among others. However, in some literature, the RESOLFT method is described as follows ( DE 10 2011 055 367 A1 , DE 10 2015 105 018 A1 ): A majority of the fluorophores are in the fluorescence-capable state Z2. By switching off or Switching signal (fluorescence prevention light), most of the fluorophores in the fluorescence-capable light state Z2 are transferred to the non-fluorescence-capable dark state.

Only a few fluorophores in the fluorescing state Z2 that are spatially far apart from one another remain on the sample surface, which can be brought into the fluorescing state Z3 by excitation light and from there emit fluorescence photons, which can be detected and contribute to imaging. If all the fluorophores are in the non-fluorescent dark state Z1, one can convert a small subset of the fluorophores from the dark state Z1 to the fluorescent light state Z2 by a small dose of turn-on signal (fluorescence-enabling light) and then proceed as described above.
In general, however, the embodiment described in the previous paragraph is understood to be the GSD method, which will be discussed in detail again in the following section and which represents only one of several embodiments of the basic RESOLFT principle.

GSD:

For this purpose, reference is made to the publications DE 10 2006 047 912 A1 , US 2011/0215258 A1 as well as [7] - [9]. In the GSD (ground state depletion) method, the de-excitation beam S3 does not de-excite from the fluorescent state Z3 to the fluorescent state Z2, but instead there is a transition from the fluorescent state Z2 to the non-fluorescent dark state Z1 (case 1) before or after the fluorescence excitation Excitation induces a transition from the fluorescent state Z3 to the non-fluorescent dark state Z1 (case 2), either before the excitation light S2 will excite the electrons from the fluorescent state Z2 to the fluorescent state Z3 (case 1) or after the excitation light S2 will excite the Excited electrons from the fluorescent state Z2 to the fluorescent state Z3.
The dark state is often a long-lived triplet state. The exemplary embodiment according to case 1 is discussed again in detail: Before the fluorophores are excited into the fluorescent state Z3, (most) fluorophores are in the fluorescent state Z2. A switching signal can cause a large part of the fluorophores in the Z2 state to enter a long-lived dark state, so that they are no longer available for fluorescence excitation. Now there are only a few fluorophores in the fluorescent bright state Z2, which are at an average distance from each other. With a suitable excitation light, these few fluorophores, which are far from one another and thus isolated from one another, are brought into the fluorescent state Z3; they return to the initial state Z2 by emitting fluorescence photons, while the emitted fluorescence photons can be detected and used for imaging.

In principle, the STED method is not limited to electronic excitation and de-excitation and transitions between electronic electron levels, but other types of excitation and de-excitation, such as between individual vibrational or rotational levels of the fluorophore molecule, also appear possible. For example, if the de-excitation process from Z3 to Z2 occurs between two vibrational levels in the same electronically excited state, then an IR photon may be emitted instead of a UV fluorescence photon; or if the fluorophore is in the Z3 state, in the form of a high vibrational state in an electronically excited state, then an IR depletion beam S3 or S4 can bring the fluorophore to a lower vibrational state in the same electronically excited state from which it can then be emission of a fluorescence photon into the electronic ground state. Instead of the normal fluorescence transitions in the STED process, X-ray fluorescence can also be used, especially since X-rays have a very short wavelength, which is advantageous for the resolution.

Also other optical or non-optical activation or excitation types, for example by means of Raman processes, elastic or inelastic impact or scattering processes with electrons, protons or other particles, are also conceivable in order to move from an energetically lower state Z1 to an energetically middle state Z2 or from to reach an energetically middle state Z2 to an energetically high state Z3. For example, the fluorophores can be brought from the Z1 state to the Z2 state or from the Z2 state to the Z3 state by means of an applied voltage or opto-acoustic processes, or the fluorophores can be brought from the Z1 state to the Z2 state or from the Z3 state by means of electroluminescence Move Z2 to state Z1 or from Z2 to Z3 or from Z3 to Z2. However, with these alternative types of excitation and de-excitation, through which the fluorophore moves from state Z3 to state Z2 in particular, the spatial resolution would be lost simply because of the missing zero point; unless one masters a process in which particles could release part of the energy at a precise location through inelastic impact or scattering processes in order to (electronically) switch the fluorophore on or off gene, in particular from state Z3 to state Z2, because highly accelerated particles such as electrons according to de Broglie have a wavelength that is far below the optical wavelengths, namely in the lower nanometer range.

Instead of zeros, one could use the minima or maxima of diffraction and interference patterns of interfering electrons or the focus of an electron beam for the precise excitation or de-excitation of the fluorophores, since the diameter of the interference fringes or the focus diameter at high particle energy due to the de Broglie relationship can be much smaller than the zero of the de-excitation light S3.

If the electrons are accelerated to high velocities in order to obtain a very short de Broglie wavelength, but as a result have too high an energy for the resonant excitation or de-excitation of the fluorophore, then the electrons can also graze incident, to only a part of delivering energy to the fluorophore. A further advantage would be that the fluorophore would not be destroyed so easily by a grazing incidence. Because of the greater mass compared to electrons, much shorter wavelengths and thus much smaller focus diameters and narrower interference fringes could be achieved according to the de Broglie relationship with protons and neutrons accelerated to high speeds; however, the danger of destroying the sample or fluorophore by the incursion of these massive particles also increases. Thus, only dead matter such as semiconductor materials could be examined, but not sensitive organic samples.

There are already a few STED methods in which the excitation light is guided through fiber optics [10] ( U.S. 2016/0170136 A1 , CN 000105241857A , CN 000104296685A , CN 000103616330A , DE 10 2007 048 135 B4 , DE 103 40 964 A1 , U.S. 2014/0369370 A1 , U.S. 2010/0142054 A1 , US2007/0025662 A1 ). However, this is not done to guide the beam or shape the beam, but to change the optical properties of the excitation beam (wavelength, spectral width, etc.) and thus optimize it for the respective special application.

Only a few STED methods have fiber optics for the excitation or emission light beam ( CN 000105467610A ). The advantage of such an arrangement is that you can also examine surfaces that are difficult or impossible to reach with a free-space optical setup, since the sample surface to be measured is either behind an undercut, in a deep bore, in holes or channels or something else covered or hidden and thus difficult to access "around a corner" or in a (chemically) aggressive or humid environment, as is often the case in materials or materials testing, or in a very bright environment or in a radiation-stressed environment where For example, the (radioactive) background radiation or other stray light that could disturb the STED measurement process.

Also sample surfaces that are covered with a medium, for example due to the examination or application, for example by water or by powder, sand, foam or water vapor with a high absorption especially for the excitation and/or emission wavelength, or which are within foreign laser radiation or a medium are located, such as within gases, plasma, mist, vapors or that are otherwise contaminated in any way, be it through smoke or other environmental conditions at the measuring workplace that are harmful or aggressive for the measurement (high temperature and/or pressure), which can be found on site (e.g. in a production facility in a factory) cannot easily be parked, cannot be examined using free-space optics, but only using fiber optics.

The disadvantage of fiber optics, however, is that, firstly, the attenuation is particularly high, especially for UV light, which is often used as high-energy excitation light (unless you use fiber optics made of pure quartz glass, which is very expensive, or of calcium fluoride, which are difficult to produce and hardly commercially available) and secondly, the optical properties of the excitation and/or emission light beam such as polarization, intensity, group or phase velocity, phase, etc. are determined by the fiber optics due to their material properties (material attenuation, dispersion, anisotropy, NLO, etc .) changed in an undesired way.

So far, no high-resolution microscopy method is known which examines sample surfaces in hard-to-reach places, for example around corners or behind undercuts (so-called two-and-a-half-dimensional structures in microtechnology) or under water or in a flowing fluid medium, e.g. in a flow tube or a microchannel. or in another hostile environment (e.g. heavily smoked or misted with water vapor, as is often found in production facilities also in microtechnology (etching, sputtering, spraying, coating, plasma treatment, etc.)) without the excitation light (often UV Radiation) or the emission light (often in the Vis/IR range) is strongly absorbed or otherwise manipulated.

Capillary hollow structures were able to establish themselves as capillary optics for focusing X-ray and neutron radiation ( DE 10 2007 049 929 A1 , DE 10 2007 049 930 A1 , WO 2008/135542 A1 , DE 10 2007 020 800 A1 ; EP 2 152 928 A1 , WO 2007/079975 A1 , WO 2007/079975 A8 , DE10 2005 063 127 A1 , U.S. 2009/138996 A1 , EP 1 969 606 A1 ).

Capillary optics/hollow capillaries/waveguides are structures that consist of one (monocapillary or monocapillary optics) or several (polycapillary, polycapillary optics or multichannel structure) spatially arranged at regular distances from each other, continuous channels or channels that are open at the ends.

In optical technologies, the beam is often guided by optical components made of glass, such as glass fibers. Capillary optics represent similar beam guidance systems or light guides, the channel of which consists of a hollow cylindrical or hollow elliptical wall made of glass. The (X-ray) light beam is reflected on the wall in the inner channel of the optics by (external or external) total reflection: For X-rays, the refractive index of matter is slightly less than one. This results from a phase velocity of the X-rays in the material that is greater than the speed of light [11]. This is discussed in more detail below. In contrast to the generally known internal or internal total reflection, the external or external total reflection is usually only known to experts in X-ray optics.

With an ellipsoidal channel shape, a particularly high beam focusing can be achieved in the second focal point when the source is placed in the first focal point. Capillary optics are manufactured from regularly arranged, uncoated glass capillaries or glass rods using various drawing and sintering processes.

At the same time, capillary optics are currently made exclusively of glass, since glass is the only material that fully meets the high requirements for the production of polycapillary structures due to its unique technical-physical and chemical properties (high radiation resistance, very good formability, optimal flow properties and easy processing). . In addition, glass has the required very low roughness. It is known from the literature that the surface roughness of flat glass is very low at rms = 0.2 nm [12].

A disadvantage of capillary optics is their relatively low efficiency, because the critical angle for external or total external reflection is very small. The reflectivity of surfaces increases as the angle of incidence becomes flatter. As already discussed above, very many solids have a refractive index of less than one for X-rays; and in the case of a refractive index less than one, total reflection can even occur at very shallow angles of incidence. For this reason, mirrors, but also capillary optics under grazing incidence, are often used in X-ray optics. As a result, only a very small proportion is transmitted through the glass wall.

Subsequent metallic coating of the capillary optics consisting of a high-Z material (material that consists of elements with a high atomic number Z high-Z element) can significantly increase the critical angle of total reflection and thus the solid angle. Significantly increased reflectivity could be successfully achieved with such an inner coating (see list in the appendix: “Own patents (Jörn Wochnowski)”).

Unevenness in the surface (surface roughness) causes undesired diffuse scattering. When the surface roughness is significantly smaller than the wavelength of the light to be reflected, there is negligible diffuse stray radiation in relation to the reflected fraction, so that reflective coatings for radiation with a long wavelength have relatively low planarity requirements. However, if the surface roughness is equal to or greater than the wavelength of the light to be reflected, there is mainly diffuse scattered radiation and only a very small proportion of reflection.

• Hierbei kommt der chemischen Gasphasenabscheidung (CVD) eine besondere Bedeutung zu. Bei der chemischen Gasphasenabscheidung (CVD) wird an der Oberfläche eines Substrates aus der Gasphase infolge einer chemischen Reaktion eine Feststoffkomponente abgeschieden. Dazu werden flüchtige Verbindungen (Precursoren) verwendet, aus denen sich bei bestimmten geeigneten Prozessparametern feste Schichten oder Partikel abscheiden. Im Gegensatz zur verallgemeinerten Definition nach Fischer wird hier aus pragmatischen Gründen eine Differenzierung der Verfahren der chemischen Gasphasenabscheidung nach den Precursorverbindungen in Organometall- (OMCVD), Metallorganische- (MOCVD), Elementorganische- (EOCVD) und Organischeelement- (OECVD) CVD-Verfahren vorgenommen.

Uncoated capillary optics are already widely used, for example in analytical instruments, but not yet in STED processes. As already indicated above, capillary optics can be subsequently coated / functionalized from the inside using suitable coating processes and thus significantly increase their performance (see list in the appendix: "Own patents (Jörn Wochnowski)"), which will be discussed in detail below:

• Chemical vapor deposition (CVD) is of particular importance here. In chemical vapor deposition (CVD), a solid component is deposited on the surface of a substrate from the gas phase as a result of a chemical reaction. For this purpose, volatile compounds (precursors) are used, from which solid layers or particles are deposited with certain suitable process parameters. Contrasted with the generalized defi According to Fischer, for pragmatic reasons, the chemical vapor deposition processes are differentiated according to the precursor compounds in organometallic (OMCVD), organometallic (MOCVD), element-organic (EOCVD) and organic element (OECVD) CVD processes.

For the substrates, such as hollow fibers, fiber bundles or multi-channel structures, the CVI (Chemical Vapor Infiltration) process is used instead of the CVD process, which, in contrast to the CVD process, enables a more uniform internal coating of these substrates.

In chemical vapor deposition, the layer growth takes place via several complex chemical and physical processes accompanied by a chemical conversion. These can be described by several sub-steps.

In step 1, the evaporated or sublimated gaseous organometallic, organometallic, element-organic or complex compound(s) are transported to the substrate to be coated as a result of flow or diffusion. The deposition can then take place in two alternative ways: In the first way, reactions take place in the gas phase by coupling in energy, with the ligands of the precursor being (partially) split off. The intermediate stages of the precursor adsorb on the substrate and continue to react analogously to the second path. In the second way, the precursor is adsorbed on the substrate without being decomposed. The ligands are then split off only on the surface by coupling in energy in subsequent surface reactions. The volatile ligands that are split off desorb from the surface and are transported away [13]. The individual deposited layer and particle systems can differ in a variety of criteria, such as their layer types, particle sizes, particle size distribution, grain boundaries, layer thicknesses, amorphity or crystallinity, topography, density, morphology, corrugation (Z or height), surface roughness, interface roughness, mechanical Stresses, disorder and relaxation, texture, lattice mismatch, adhesion, chemical compositions, and purities differ and vary widely.

• • Substratwahl
• • Prozessdrücke
• • Substrattemperaturen
• • Wachstumsraten
• • Verunreinigungen
• • (Thermische) Nachbehandlung(en)
• • Precursor(en)auswahl

The type of layer structures deposited can be significantly influenced by a suitable choice of the following process conditions:

• • Choice of substrate
• • Process pressures
• • substrate temperatures
• • growth rates
• • Impurities
• • (Thermal) post-treatment(s)
• • Precursor(s) selection

Depending on the coating substrates used, a distinction is made between so-called "growth surfaces" and "non-growth surfaces". There is a close correlation between the substrates used in each case and the layer structures deposited on them, some of which can fundamentally differ from one another with regard to their morphology, topography and their physical or chemical compositions.

Therefore, knowledge of the type and selection of the suitable coating substrate (inside of the hollow capillary) is just as important for the layer structure as the selection of the precursors, but also the process conditions such as the substrate temperature.

As already mentioned above, unevenness in the surface (surface roughness) causes undesired diffuse scattering. When the surface roughness is significantly smaller than the wavelength of the light to be reflected, there is only negligible diffuse stray radiation in contrast to the reflected portion, so that reflective coatings for radiation with a long wavelength have relatively low planarity requirements. However, if the surface roughness is equal to or greater than the wavelength of the light to be reflected, there is mainly diffuse scattered radiation and only a very small proportion of reflection.

For this reason, surfaces that are absolutely planar in visible light are usually far too rough for extremely short-wave light such as UV or X-ray light. In the case of hard X-rays, for example, the wavelength is smaller than the distance between the atoms in the solid body, which means that the atomic structures of the coating are important. Subsequent interior coatings for the reflection of hard X-rays must therefore have surface qualities and qualities that are comparable to or better than those of uncoated glass surfaces in order to ensure low-loss transport of the radiation.

• Beispielsweise ist für EUV-Strahlung mit der Wellenlänge λ= 13,5 nm an einer Grenzfläche aus Quarzglas bei einem typischen Einfallswinkel von 80° bezogen auf das Flächennormal von einem rechnerischen Reflexionsgrad von 30 % auszugehen. Der Reflexionsgrad kann sich auf beispielsweise ca. 75 % steigern lassen, wenn die Oberfläche mit einer idealisierten Metallschicht beispielsweise bestehend aus Palladium modifiziert wird. Unter idealen Bedingungen wird daher eine Steigerung der Effizienz bei der Abbildung eines EUV-Strahls in der STED-Mikroskopie um mehr als einen Faktor zwei in Aussicht gestellt.

The coating materials palladium, tungsten and platinum and their compounds for the optical refinement of capillary optics (coating of the inner wall) turned out to be particularly suitable for STED microscopy:

• For example, for EUV radiation with a wavelength of λ= 13.5 nm at a quartz glass interface at a typical angle of incidence of 80° relative to the surface normal, a calculated degree of reflection of 30% can be assumed. The degree of reflection can be increased to about 75%, for example, if the surface is modified with an idealized metal layer consisting of palladium, for example. Under ideal conditions, an increase in the efficiency of imaging an EUV beam in STED microscopy by more than a factor of two is therefore promised.

Task:

It is therefore the object of this invention to provide a device and a method which combines the advantages of the already known high-resolution microscopy devices and methods such as STED (resolution beyond the classic resolution limit) with those of fiber optics, in particular waveguides (sufficiently suitable beam guidance and shaping to reach places on the sample surface that are difficult to access and/or in metrologically difficult surroundings), without however having to accept the usual disadvantages of fiber optics (in particular high (material) attenuation for high-energy radiation and dispersion and possibly no polarization maintenance).

General principle solution:

Therefore, optics that are equivalent to fiber optics are required, which combine the advantages of solid core fiber optics (excitation and detection in places that are otherwise very difficult to reach or in an environment that is problematic from a metrological point of view) and the advantages of a free-space optical arrangement (no negative manipulation of the excitation and emission radiation in terms of their optical properties, in particular attenuation, dispersion and polarization) are combined with one another.

As an alternative to solid-core fiber optics, there are hollow fiber optics in the form of capillary optics, such as the inner sides of the (micro) hollow capillaries that are finished in optical quality high-resolution microscopy device and method are to be used.

However, the inside must be modified accordingly so that the high-energy UV light or X-rays can be guided and guided (and possibly shaped) with as little loss as possible without the properties of the radiation being adversely affected (of course, apart from the desired beam guidance and -forming).

To do this, however, the inside of the waveguide must be provided with a suitable coating which, firstly, can conduct the high-energy excitation light with as little loss as possible and without any negative effects, and secondly, which is also flexible enough to reach less accessible areas and to apply excitation light to them or to cool them down to capture the emission light.

A suitable coating process is presented below, with which the inside of the waveguide can be coated using a (MO)CVD process.

With this (MO)CVD method, the physical-chemical and/or optical-functional properties of the layer can be adjusted in a targeted and controlled manner. For this purpose, a suitable capillary hollow structure, for example made of quartz glass, is first selected and provided. These can also be inexpensive, commercially acquirable mass-produced goods. This capillary hollow structure is then cut to size, cleaned and subjected to other necessary preparation procedures and ultimately pushed into the recipient of a (MO)CVD system on a substrate holder.

After the recipient has been flushed and evacuated, the appropriate organometallic precursor materials are introduced into the recipient under the required process conditions. The desired process parameters (temperature, pressure, flow rate, etc.) are then set. Since the organometallic precursor materials are thermodynamically very unstable, they already decompose under normal conditions (room temperature, normal pressure) in that the organometallic residues (partly) fall off the metallic central particle, so that the metallic central particle in the gas phase and/or on the sample surface (inner wall of the Quartz capillary hollow structure) are released. The defragmentation and separation processes discussed in the prior art then take place. After a metal layer has been deposited in the desired thickness, i.e. after a certain coating time, the deposition process is interrupted by pumping the precursor materials out of the recipient.

After flushing with an inert gas (e.g. high-purity nitrogen or inert gas), the recipient can be opened and the sample taken. A quality check may be completed of the inner-coated hollow optical waveguide produced in this way.

So that the excitation light can be coupled into the inner-coated hollow fiber optic cable produced in this way from one side and out again on the other side, suitable optics must be developed at the ends of the hollow fiber for STED in order to ensure that the radiation guided through them is coupled in and out with as little loss as possible (see list in the appendix: "Own patents (Jörn Wochnowski)").

In addition, with such a light-guiding optics such as a waveguide, there is the possibility of temperature control on the inside. This temperature control can take place, for example, by means of a heated or cooled carrier gas flow, in order to keep the inner coatings and thus the substrate surface (hollow optical waveguide) constantly at the desired temperature. From the current state of knowledge, it cannot be ruled out that there is a (slight) dependence of the reflection behavior on the temperature, which can be exploited here according to the invention.

All of the above applies not only to the measurement / characterization of a sample surface, but also to the "fluorescence-based" nanostructuring of the sample surface, as for example in the German patent application DE 10 2015 015 497 A1 has been described.

For this purpose, the excitation beam is then sent through the waveguide in order to induce the emission of one or a few fluorescence photons on a sample surface that is difficult to access, which can structure, modify or otherwise change the immediate atomic and/or molecular environment, with a Resolution that is far below the classical Abbe resolution limit. Although it is theoretically possible to process sample surfaces with a so-called Pendry lens consisting of a material with a negative refractive index with a resolution that is beyond the classic resolution limit according to Abbe, this only applies to points on the sample surface that are easily accessible using free-space optics.

Concrete examples

• Die Vorrichtung umfasst eine Lichtquelle 1 mit einem Laser 2 zum Aussenden von Anregungslicht, um die damit zu beaufschlagenden Fluorophore auf der Probenoberfläche von dem fluoreszenten Zustand Z2 auf den fluoreszierenden Zustand Z3 zu bringen, und einem weiteren Laser 3 zum Aussenden von Stimulationslicht bzw. Abregungslicht (Fluoreszenzverhinderungslicht), um die Fluorophore auf der Probenoberfläche vom fluoreszierenden Zustand Z3 wieder zurück auf den fluoreszenten Zustand Z2 zu transferieren. Dabei kann der Stimulationslichtstrahl auch anders, beispielsweise mittels einer Vortex-Phasenplatte oder eines SPM (Spatial Phase Modulator) oder mittels anderer optischer Komponenten, erzeugt werden, wie beispielsweise in DE 20 2009 007 250 U1 , DE 10 2019 008 304 B3 , DE 10 2011 055 367 A1 , DE 10 2018 132 875 A1 , DE 10 2009 056 250 A1 , WO 2020/128106 A1 beschrieben. Die Lichtquelle 1 umfasst zu den Lasern 2 und 3 zusätzlich noch Linsen 12 und 13 sowie eine Blende 14. Mit der Linse 12 wird der von dem Laser 2 kommende Laserstrahl auf die Blende 14 fokussiert.

The STED structure required for the subject matter according to the invention initially resembles that which has already been described in the prior art [1]-[9], in particular in the German patent application DE 44 16 558 A1 , especially there 1 , as has been described below in a slightly modified manner; however, if necessary, modifications are also possible:

• The device comprises a light source 1 with a laser 2 for emitting excitation light in order to bring the fluorophores on the sample surface to which it is applied from the fluorescent state Z2 to the fluorescent state Z3, and a further laser 3 for emitting stimulation light or de-excitation light ( fluorescence preventing light) to transfer the fluorophores on the sample surface from the fluorescent state Z3 back to the fluorescent state Z2. The stimulation light beam can also be generated differently, for example by means of a vortex phase plate or an SPM (Spatial Phase Modulator) or by means of other optical components, such as in DE 20 2009 007 250 U1 , DE 10 2019 008 304 B3 , DE 10 2011 055 367 A1 , DE 10 2018 132 875 A1 , DE 10 2009 056 250 A1 , WO 2020/128106 A1 described. In addition to the lasers 2 and 3 , the light source 1 also includes lenses 12 and 13 and a diaphragm 14 . The laser beam coming from the laser 2 is focused onto the diaphragm 14 with the lens 12 .

Furthermore, dichroic mirrors 4 and 5 and a lens 6 for beam guidance and shaping are installed, so that the excitation light and stimulation light coming from the lasers 2 and 3 can be directed onto a sample point 7 of the sample 8 and focused.

The lens 13 serves to adjust the divergence of the excitation light beam and the stimulation light beams so that they can be focused in the same plane using the objective lens 6 . Pinhole diaphragms are usually used as diaphragms. A beam splitter 23 and a mirror 24 for dividing the beam coming from the laser 3 into two stimulation light beams 17 are arranged behind the laser 3 . The arrangement is chosen so that the excitation light beam and the stimulation light beams impinge on the mirror 4 in such a way that the intensity distributions of the beams after deflection at the mirror 5 and passing through the lens 6 in the focal area of the lens 6 partially coincide. The sample 8 is placed on a positioning table 10 . The sample point 7 has a spatial extent, which is an area extent here. A detector 9 is arranged to detect the emission light emitted by the sample 8, which passes through the dichroic mirror 5 and is thus separated from the excitation light. Between the light source 1 and the lens 6 is a beam raster device 11 for controlled abrasion tern of the sample 8 provided with the excitation light and the stimulation light.

In contrast to that in the German patent application DE 44 16 558 A1 described completely free-space-optically constructed STED device, the device according to the invention is equipped with micro hollow capillaries in the light paths in order to guide and/or shape the excitation and de-excitation light radiation as well as the emission light radiation as is necessary according to the experimental or application-related requirements in order to achieve an optimal to achieve result. For this purpose, in particular the beam path between the mirror 5 and the sample 8, or at least on the path just before the sample surface, the free-space optical beam grid device 11 can be replaced by a grid device for fiber optics.

Concrete examples of deposition processes:

Specific capillary hollow structures (material: quartz glass, geometry such as diameter, cross-section geometry, manufacturer, pre-treatment), specific precursor materials: which MO compounds and which metals are to be deposited, information on the system such as recipients (material, size, no energy coupling), information on the deposition process such as process parameters (temperature, pressure, flow rate, coating time, how much volume of precursor material has to be introduced).

With regard to the specific process parameters, reference is also made to the publications in the list of our own patent literature.

The challenge here is that the hollow light waveguides (waveguides) are designed in such a way that the zero point of the fluorescence prevention light is retained or at least a zero point of the fluorescence prevention light is created at the correct point when it exits the waveguide, as this is essential for the STED/RESOLFT process is. This can be achieved by implementing suitable micro-optics at the front and rear of the ends of the waveguide, so that the de-excitation or fluorescence-preventing radiation is guided and shaped in such a way that this zero point is created at the exit end of the waveguide, for example by imaging. It is also conceivable that a vortex phase plate is implemented at the exit end of the waveguide.

However, the waveguide can also be designed in such a way that mode fields can form in this waveguide which have an intensity minimum in the center. For this purpose, the waveguide must have a suitable geometry. However, the prerequisite is that the waveguide is suitable for guiding and shaping the beam of UV light, i.e. the inner coating of the waveguide must be designed in such a way that, firstly, the degree of reflection is particularly good for this wavelength range so that the attenuation is not too high, and secondly that other requirements such as polarization maintenance, etc. are guaranteed so that the coherence of the light guided through the waveguide is maintained and the light thus remains capable of interference at the exit end of the hollow fiber.

In order to increase the degree of reflection, the inner coating can also be constructed in the form of a dielectric mirror layer system. For this purpose, several layers with different refractive indices and a precisely adjusted layer thickness are deposited one on top of the other, so that a multi-layer system of layers deposited one on top of the other is created, which can reflect light of a certain wavelength and for a certain angle of incidence (mode) almost 100%. Instead of or in addition to the multi-layer systems, gradient layers are also conceivable or no complete coatings that do not cover the entire surface of the inside of the microcapillary, but only in strips or even at certain points, so that other multi-layer systems for other wavelengths can be used at other points on the inside of the capillary and/or other modes (angles of incidence) can be implemented. As a result, several wavelengths can then be guided simultaneously through the hollow light waveguide.

It must also be investigated to what extent curvature of the waveguide can influence the intensity distribution within the guided light field, in particular the zero point at the exit end of the waveguide.

The hollow light waveguide, in particular the substrate material, should therefore be made of a (highly) flexible, (highly) elastic and mechanically resilient material in order to have the property of elastic deformation, so that it can be bent reversibly and flexibly, firstly to possibly under to be able to scan the sample surface buried in a liquid layer more easily and, secondly, to be able to access hidden surfaces such as undercuts.

• Zunächst ist ein exzellentes Reflexionsvermögen, abhängig von der Wellenlänge, bevorzugt im UV- oder kürzerwelligen Spektralbereich, erforderlich. Hierzu muss die Rauheit der Schichtoberfläche kleiner oder gleich der Wellenlänge der eingesetzten Strahlung sein, damit die Reflexion nicht diffus und ungerichtet wird. Vorzugsweise bestehen die Beschichtungen aus nicht oxidierbaren Edelmetallen, was eine hohe Langzeitbeständigkeit sicherstellt. Bevorzugt sind Innenbeschichtungen so auszugestalten, um die Dispersionen zu minimieren. Hierzu zählen u.a. auch die bereits oben genannten dielektrischen Schichten.

For this purpose, the layers must be produced as follows so that they have the following functional-optical properties:

• First of all, excellent reflectivity is required, depending on the wavelength, preferably in the UV or shorter wavelength spectral range. For this purpose, the roughness of the Layer surface must be smaller than or equal to the wavelength of the radiation used, so that the reflection is not diffuse and undirected. The coatings are preferably made of non-oxidizable precious metals, which ensures high long-term durability. Inner coatings are preferably designed in such a way as to minimize dispersion. These include, inter alia, the dielectric layers already mentioned above.

In a particular embodiment, the hollow capillaries are preferably made of (quartz) glass, which is actually a highly brittle and mechanically less resilient material because it is not very flexible and therefore inflexible, provided with an outer coating by means of a so-called cladding a material other than glass, such as preferably a metal-containing or ceramic material or particularly preferably a polymeric material, in order to ensure, among other things, the ultra-high vacuum stability and thus the loss-free light and energy transmission at the vacuum light speed even with (strong) bending. The advantage of this embodiment of such a hollow capillary provided with the outer coating (cladding) described above is that it eliminates the above-mentioned disadvantages and thus the highly flexible and highly flexible and mechanically very resilient hollow optical waveguides required for applications in STED microscopy can be provided .

Outlook:

Another advantage of the waveguide compared to solid-core fiber optics is that the waveguide can also be used to transport particle radiation such as electron, proton, ion or neutron radiation. As already explained in the "Prior art" chapter, it is also conceivable to use excitation radiation and/or STED radiation (fluorescence prevention or stimulation radiation) in the form of particle or corpuscular radiation (electron, proton, neutron or ion radiation, etc.) ) to act on the sample surface to be examined. This is not possible with solid core fiber optics, whether made of glass or polymer. Furthermore, by means of the internally coated waveguide according to the invention, the particle radiation can not only be directed and guided, but also shaped and its radiation-optical properties can be influenced. Externally applied (static or time-varying/dynamic, homogeneous or inhomogeneous) electrical, magnetic and/or electromagnetic fields may be necessary for this.

Literature:

• [1] Stefan W. Hell: New resolution law in light microscopy enables images with unprecedented sharpness. In: Activity report of the Max Planck Society, 2005; Biophys J. 2011 Apr 20; 100 (8): L43-L45 , Stefan W. Hell, Marcus Dyba, Stefan Jacobs: Concepts for nanoscale resolution in fluorescence microscopy. In: Current Opinion in Neurobiology, Vol. 14, 2004, pages 599-609 ; Stefan W. Hell: Microscopy and its focal switch. In: Nature Methods, Vol. 1, Jan 2009, pages 24 - 32 ; Stefan W. Hell: Nanoscopy with focused light. In: Physics Journal, Vol. 12, pages 47-53 ; DE 10 2015 104 368 ,
• [2] Stefan W. Hell and Jan Wichmann: Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy. In: OpticsLetters. 19, No. 11, 1994, pp. 780-782 , doi:10.1364/OL. 19.000780; Thomas A. Klar, Stefan W. Hell: Subdiffraction resolution in far-field fluorescence microscopy. In: OpticsLetters. Vol. 24, No. 14, 1999, pp. 954-956 , doi:10.1364/OL.24.000954.
• [3] Stefan W. Hell: Toward fluorescence nanoscopy. In: Nature Biotechnology, Vol. 11, Nov 2003, pages 1347-1355 ; SW Hell, S Jakobs, L Kastrup: Imaging and writing at the nanoscale with focused visible light through saturable optical transitions. In: Appl. physics A, Vol. 77, 2003, pages 859 - 560
• [4] DE 44 16 558 A1 , US 2011/0215258 A1 , US 2007/0023686 A1 , WO 2011/090710 A2 , DE 10 2015 116 023
• [5] DE 103 25 459 A1 , DE 103 25 460 A1
• [6] Jörg Bewersdorf, Christian Eggeling, Thomas A. Sure: Abbe tricked. In: Physik Journal, Vol. 13, No. 12, 2014, pp. 23 - 27; Christian Schumann, Annette Kraegeloh: A sharp look at nanoparticles. In: Physics Journal, Vol. 10, No. 4, 2011 , pages 27 - 32
• [7] Stefan W. Hell, M. Kroug: Ground-state-depletion fluorescence microscopy: a concept for breaking the diffraction resolution limit. In: Applied Physics B: Lasers and Optics. Vol. 60, No. 5, 1995, pp. 495-497 , doi:10.1007/BF01081333.
• [8th] Stefan Bretschneider, Christian Eggeling, Stefan W. Hell: Breaking the diffraction barrier in fluorescence microscopy by optical shelving. In: Physical Review Letters. Vol. 98, No. 5, 2007, p. 218103 , doi:10.1103/PhysRevLett.98.218103.
• [9] Volker Dose: Peer review. In: EPL, A Letters Journal Exploring the Frontiers of Physics. Vol. 89, 2009, doi:10.1209/0295-5075/86/10000.
• [10] Chetan Poudel, Clemens F. Kaminski, Supercontinuum radiation in fluorescence microscopy and biomedical imaging applications, Journal of the Optical Society of America B Vol. 36, Issue 2, pp. A139-A153 (2019) • https://doi.org/10.1364/JOSAB.36.00A139
• [11] On the Internet: <URL: https://de.wikipedia.org/wiki/R%C3%B6ntgenoptik>. researched on 06/25/2021
• [12] E. Rädlein, WOPAG - University of Bayreuth, http://www.wopag.uni-bayreuth.de, 2005.
• [13] MJ Hamden-Smith, T.T. Kodas, Chem. Vap. Deposition, 1, 8-23, 1995.

Own patent literature (J. Wochnowski):

• 1. Hollow optical waveguides coated on the inside, methods for their production and their use (Hollow waveguide used in medicine and in structural analysis comprises a channel structure having an inner coating with a specified thickness) patents: DE 10 2007 049 929 A1 2009-04-23; DE200710049929 20071018 ; DE 102007049929 (B4 ) 2011-05-05
• 2. Surface-modified cavity structures, processes for their production and their use (surface-modified structures, useful eg in optical or catalytic applications, comprise substrate, eg glass, silicate primary coating and secondary coating, eg metal) Patents: DE 10 2007 049 930 A1 2009-04-23; DE200710049930 20071018 ; DE 10 2007 049 930 B4 2011-04-28
• 3. Modified multichannel structures and their use (Modified multichannel structures and their production and use) patents: WO 2008/135542 A1 2008-11-13; WO2008 EP55458 20080505 ; DE 10 2007 020 800 A1 2008-11-06; DE 10 2007 020 800 A1 2011-03-03; DE200710020800 20070503 ; DE10 2007 020 800 B4 ; EP 2 152 928 A1
• 4. Fabrication of microtips and nanotips for use in scanning field microscopy Patents: WO 2007/079975 A1 2007-07-19; WO2006 EP12588 20061228 ; WO 2007/079975 A8 ; DE200510063127 20051230 ; DE 10 2005 063 127 A1 2007-08-23; DE 10 2005 063 127 B3 ; U.S. 2009/138996 A1 ; EP 1 969 606 A1

QUOTES INCLUDED IN DESCRIPTION

This list of documents cited by the applicant was generated automatically and is included solely for the better information of the reader. The list is not part of the German patent or utility model application. The DPMA assumes no liability for any errors or omissions.

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• Stefan Bretschneider, Christian Eggeling, Stefan W. Hell: Breaking the diffraction barrier in fluorescence microscopy by optical shelving. In: Physical Review Letters. Vol. 98, No. 5, 2007, p. 218103 [0073]
• Chetan Poudel, Clemens F. Kaminski, Supercontinuum radiation in fluorescence microscopy and biomedical imaging applications, Journal of the Optical Society of America B Vol. 36, Issue 2, pp. A139-A153 (2019) [0073]

Claims (12)

High-resolution fluorescence-based microscopy device such as RESOLFT, STED or GSD, in which the excitation light and the emission light are guided by means of a hollow fiber or a hollow optical waveguide (waveguide) in the form of a hollow capillary, in particular a hollow microcapillary, for passive beam guidance and beam shaping towards the sample surface to be examined and which is difficult to access. and be guided away. device after claim 1 , In which the used (s) hollow capillary (s) according to the claim 1 is an internally coated functionalized hollow fiber optic cable (FHLWL), in which the channel structure (a) consists of an amorphous solid, preferably glass, and/or a macromolecular material, preferably a polymer, and the internal coating (b) is an element other than carbon from the first to fifth main group or a subgroup of the periodic table of elements, wherein the inner coating has a higher optical density than the channel structure, and in which the inner coating has at least one metamaterial and/or a metal, with a metal being selected from the group for the inner coating comprising Be, Ni, Al, Pt, Cu, Pd, Ag, Au, W, Re, Ir, Os, Mo, Pb, Bi and U, where the elements are Au, Ag, W, Pb, Pt, Pd, Ni and A1 are particularly preferred and/or in which the inner coating contains a metal with an atomic number Z>10, in particular Z>21, and the inner coating consists of metal layers and/or metal particles, preferably amorphous metal layers and/or metal particles, in particular amorphous Metal layers and particularly preferably closed metal layers, the degree of coverage of the inner coating of the channel structure being at least 50%, in particular at least 90% and particularly preferably at least 99%. Internally coated functionalized hollow fiber optic cable (FHLWL), characterized in that it has the following features: External coating by means of a cladding, the cladding being a material other than glass, such as preferably a metal-containing or ceramic material or particularly preferably a polymeric material, in order to ensure, among other things, the ultra-high vacuum stability and thus the loss-free transmission of light and energy at the vacuum speed of light even at ( strong) bending, so that the functionalized hollow fiber optic cable (FHLWL) with a cladding and an inner coating is flexible and particularly preferably highly flexible and has a significantly improved long-term stability compared to an uncoated hollow fiber optic cable (HLWL), and the outer coating of the inner-coated functionalized hollow fiber optic cable (FHLWL) has at least carbon, silicon and/or a metal as an element, and the cavity of the internally coated functionalized hollow fiber optic cable (FHLWL) is either evacuated or filled with a (liquid, vaporous or gaseous) medium, the internally coated functionalized hollow fiber optic cable (FHLWL) additionally has the following additional features: a) one- or two-sided sealing at the ends, particularly preferably by a suitable optically transparent material or by suitable micro-optics such as a light-refractive (refractive) body (e.g. optical lens) or by a reflective element or component (e.g. mirroring or dielectric Mirroring as in a laser) or by a diffractive element or component (e.g. grating or prism) or by another optical element or component b) FHLWL preferably evacuated with a vacuum and particularly preferably with an ultra-high vacuum and/or c) FHLWL filled with a material, preferably with an energy-, light- and/or mass-transparent substance filled with FHLWL, and particularly preferably with an (inert) gas or protective gas at room temperature, such as nitrogen or argon Process for producing an inner coating on the inside of a channel structure and/or a functionalized hollow fiber optic cable (FHLWL) according to the previous one claim 3 , in which the application of the inner coating by a thermal, photolytic, plasma technological and / or gas-phase process such as chemical vapor deposition (CVD) or chemical vapor phase infiltration (CVI) and combinations of these processes (so-called hybrid or multi-process) by the following steps takes place: (a) evacuation of the channel structure on one or both sides, with a pressure gradient being built up between the ends of the channel structure, (b) evaporation or sublimation of the precursor compound(s) and transport through the channel structure by the pressure gradient that has been built up and/or by a supporting one carrier gas flow, and (c) depositing an inner coating at the desired location from the related precursor material by supplying energy, or in which the inner coating is formed by chemical vapor deposition of organometallic compounds (OMCVD), by chemical vapor deposition of organometallic compounds (MOCVD), by the chemical vapor deposition of organoelement compounds (OECVD), by which chemical vapor deposition of element organic compounds (EOCVD) or related chemical vapor infiltration processes such as OMCVI, MOCVI, OECVI or EOCVI or related vapor phase epitaxy processes such as OMV-PE, MOV-PE, OEV-PE or EOV-PE or by atomic layer deposition (ALD) is deposited on the inside of the channel structure, or in which the inner coating is deposited by a thermal process, photolytic process, a plasma-technological and/or a liquid-phase process (Chemical Liquid Deposition, CLD) and combinations of these processes (hybrid or multi-process ) is deposited on the inside of the channel structure and which comprises the following steps: (d) the precursor compound or compounds are dissolved in a suitable solvent; alternatively, the precursor(s) can be melted; the transport of the precursor compound(s) into the channel structure is then carried out by the capillary forces of the channel structure (e) deposition of an inner coating at the desired location from the precursor material dissolved in the liquid phase by supplying energy (f) removal of the solvent, for example by evacuation and /or Evaporation, or in which the inner coating is produced by Organometallic Chemical Liquid Deposition (OMCLD), Organometallic Chemical Liquid Deposition (MOCLD), Organoelemental Chemical Liquid Deposition (OECLD), Organic Elemental Chemical Liquid Deposition (EOCLD) or their related liquid phase chemical infiltration processes such as OMCLI, MOCLI, OECLI or EOCLI or their related liquid phase epitaxy processes such as OML-PE, MOL-PE, OEL-PE or EOL-PE is deposited on the inside of the channel structure, or in which the inner coating is deposited on the inside of the channel structure by a thermal process, a photolytic process, a plasma-technological and/or a solid-phase process and combinations of these processes (hybrid or multi-process) and comprises the following steps: (g) the precursor compound or compounds are introduced mechanically into the channel structure as a solid, for example (h) deposition of an inner coating at the desired location from the solid precursor material by supplying energy until the precursor material has completely decomposed and deposition of an inner coating (i) removal of the gaseous decomposition products of the precursor material, for example by evacuation and /or evaporation, or in which the inner coating is produced by solid phase chemical deposition of organometallic compounds (OMCSD), by solid phase chemical deposition of organometallic compounds (MOCSD), by solid phase chemical deposition of organoelement compounds (OECSD), by solid phase chemical deposition of organoelement compounds (EOCSD) or their related methods of solid phase chemical infiltration such as OMCSI, MOCSI, OECSI or EOCSI or their related methods of liquid phase epitaxy such as OMS-PE, MOS-PE, OES-PE or EOS-PE is deposited on the inside of the channel structure, and/or in which the inner surface of the channel structure is activated before said steps (a), (d) or (g), and in which in particular the inner surface of the channel structure is thermally activated before steps (a), (d) or (g), and/ or in which, in particular, the inner surface of the channel structure is activated before steps (a), (d) or (g) by passing molecular oxygen and/or ozone over it, and/or in which the inner surface of the channel structure is activated before said step (a) , (d) or (g) is cleaned, and/or in particular in which the inner surface of the channel structure is cleaned before steps (a), (d) or (g) by plasma treatment, chemical processes and/or evacuation with simultaneous heating, and/or in which the inner surface of the channel structure is evacuated and particularly preferably ultra-high evacuated before steps (c), (f) or (i), and/or in particular in which the inner surface of the channel structure in said steps (c), ( f) or (i) a minimum pressure of less than 10 ^-2 mbar, and preferably less than 10 ^-3 mbar and particularly preferably less than 10 ^-5 mbar, is set on one or both ends, preferably, and/or in which the Channel structure remains evacuated after the steps (c), (f) or (i) mentioned and particularly preferably ultra-highly evacuated, and/or in which the functionalized hollow light waveguide (FHLWL) is filled with a suitable medium such as protective gases, which have a high light, mass and exhibit energy transparency such as nitrogen or argon, are (permanently) filled, with the channel structure being filled after the coating process has taken place according to steps (c), (f) or (i) with suitable materials such as protective gases, for example, which have a high light, exhibit mass and energy transparency such as nitrogen or argon, is (permanently) filled, and/or in which the inner surface of the channel structure in said steps (b), (e) or (h) is exposed to a heat or radiant heater, an oven, a laser, microwave radiation and/or plasma, and/or which also includes the implementation of in-situ process analysis and process control, and/or in which the precursor material is a sublimable material, and/or in which the precursor material is an in a solvent-soluble material, and/or in particular in which the precursor material contains an element other than carbon from the first to fifth main groups or a subgroup of the periodic table of the elements, as well as organic groups and/or one or more carbonyl ligands which are chemically directly and/or are bonded to the respective element via an element of the fifth or sixth main group, and/or in which the precursor material is an organometallic, an organometallic or an organic element compound which has at least one metal-carbon bond, or is a complex or coordination compound , which has an organic ligand and/or carbonyl, and/or in which the complex or coordination compound contains a ligand selected from the group consisting of carbonyl, hexafluoroacetylacetonato and acetylacetonato, and/or in which the precursor material comprises one or more elements the group consisting of Be, Ni, Al, Pt, Cu, Ir, Mo, Ag, W, Re, Ir, Os, Au, Pb, Bi and U, where the elements Ni, Al, Ag, Au, W, Pb, Cu, Pt, Bi and U are particularly preferred, and/or in which the precursor material is selected from the group consisting of: trimethylaluminum triethylaluminum bis-(1,1,1,5,5,5-hexafluoro-2, 4-pentadionato)palladium(II) tetracarbonylnickel nickelocene 2,2,6,6-tetramethyl-3,5-heptanedionatosilver(II) tris(2,2,6,6-tetramethyl-3,5-heptanedionato)bismuth(III ) tetraethyl lead bis(2,2,6,6-tetramethyl-3,5-heptanedionato)lead(II) uranium hexafluoride uranocene uranium acetate tungsten hexacarbonyl tungsten hexahalides dimethyl(hexafluoroacetylacetonato)gold tetrakis(triphenylphosphine)platinum(0) bis(hexafluoroacetylacetonato)platinum(II) bis(acetylacetonato)platinum(II) dimethyl(1,5-cyclooctadiene)platinum(II) methyl(triphenylphosphine)gold(i) and bis(1,1,1,5,5,5-hexafluoro-2,4- pentadionato)-lead(II) Internally coated functionalized hollow fiber optic cable (FHLWL), characterized in that it has the following features: internally coated functionalized hollow fiber optic cable (FHLWL), in which the cavity is either evacuated or filled with a (liquid or preferably a vapor or gaseous) medium, and methods for the closure of any (functionalized) hollow fiber optic cable (HLINUFHLWL) or a functionalized hollow fiber optic cable (FHLWL) according to one or more of Claims 1 until 4 preferably at both ends with suitable optically transparent materials and/or at one or preferably at both ends with suitable micro-optics. Internally coated functionalized hollow fiber optic cable (FHLWL), characterized in that it is filled with a laser-active gas or gas mixture with a suitable mixing ratio (e.g. helium and neon, excimer or exciplex filling) and on one side with a normal dielectric mirror and on the other side sealed with a semi-transparent dielectric mirror and an energy source for exciting a laser process taking place in the laser-active gas filling. Use of any hollow fiber optic cable (HLWL/FHLWL) functionalized by an inner coating (internally coated) or a functionalized hollow fiber optic cable (FHLWL), characterized in that it can be used as a light, mass and energy waveguide for the purpose of signal, data and information, mass - and energy transfer is used, whereby the FHLWL has at least the following two characteristics: a) a channel structure and b) an inner coating on the inner surface of the channel structure, wherein the selected material of the channel structure a) must differ from the material of the inner coating and b) the inner coating improves the signal, data and information, mass and energy transmission, wherein a significant improvement is preferred and substantial improvement is particularly preferred. Use of any hollow fiber optic cable (HLWL/FHLWL) functionalized by an inner coating (internally coated) or a functionalized hollow fiber optic cable (FHLWL), characterized in that it can be used as a light, mass and energy waveguide for the purpose of signal, data and information, mass - and energy transfer is used, in which the channel structure is a mono-channel structure or a multi-channel structure, for example selected from the group consisting of mono- or poly-capillaries, compound lenses made of individual mono- and / or poly-capillaries, monolithic lenses also made of individual mono- and /or polycapillaries, photonic crystals and monolithic integral microlenses. Device according to claims 1 and 2 , In which the hollow optical waveguide according to claims 3 and 5 until 6 has a temperature control element for precisely adjustable temperature control. Method for energy, mass, data and information transmission in any (functionalized) hollow fiber optic cable (FHLWL / HLWL) or a functionalized hollow fiber optic cable (FHLWL), in which the transmission is carried out by coupling and decoupling electromagnetic radiation (light) by means of a - And decoupling according to claim 3 takes place, and in which the transmission takes place by monochromatic and/or preferably by polychromatic electromagnetic radiation, and in which the transmission preferably takes place by (high-energy) electromagnetic radiation, Vis, IR, HF and/or microwave radiation also being particularly preferred here are, while the radiation with a wavelength of less than 380 nm such as UV or X-rays is also particularly preferred, or in which the transmission by particle radiation, particularly preferably neutron radiation, and / or γ-radiation takes place, and / or in which the Transmission occurs simultaneously by different types of particle radiation, or in which transmission occurs simultaneously by polychromatic electromagnetic radiation and particle radiation, and/or in which transmission occurs by (pulsed or continuous) LASER radiation, and/or in which transmission occurs simultaneously by monochromatic LASER radiation takes place, and/or in which the transfer takes place simultaneously by polychromatic LASER radiation, for example by a white-light LASER, and/or in which the transfer takes place simultaneously by electromagnetic radiation and/or by particle radiation and/or LASER radiation, and/or in which the transfer takes place after claim 2 takes place (almost) without dispersion, and/or in which the transmission takes place at more than half the speed of light, preferably more than 2/3 of the speed of light and particularly preferably at the speed of light in a vacuum. Use of any hollow optical waveguide (HLWL/FHLWL) functionalized (internally coated) by an inner coating or a functionalized hollow optical waveguide (FHLWL) as fiber optics in the form of a hollow capillary, in particular a micro hollow capillary, in order to be used in a high-resolution, fluorescence-based microscopy device and/or method such as RESOLFT, STED or GSD to guide and shape the excitation light in order to apply it to a sample surface to be examined and which is difficult to access, possibly in an ambient atmosphere that is problematic for the measuring process, and to guide the fluorescent light then emitted away from the point to be examined on the sample surface, possibly to shape it and a supply detector. Use of any hollow optical waveguide (HLINUFHLWL) or functionalized hollow optical waveguide (FHLWL) functionalized by an inner coating (internally coated) for the purpose of use as a component or (connecting) component in applications in the field of microscopy, for example the • RESOLFT, • STED or • GSD