Vn filter

These filters provide excellent transmission of the desired emission wavelength (>90%), with a sharp spectral cutoff and low transmission at other wavelengths (<0.001%). Click on the icons above to view product-specific transmission data. For additional fluorophore compatability, please see the Fluorophores tab.

Since standard fluorescence imaging applications generally incorporate three different filters (i.e., one excitation, one emission, and one dichroic filter) to maximize the signal-to-noise ratio, Thorlabs offers these filters as a set at a savings over purchasing them separately.

The metal used in an electron microscope is tungsten. A high voltage current is applied which results in the excitation of the electrons in the form of a continuous stream that is used as a beam of light. The lenses used in the electron microscope are magnetic coils. These magnetic coils are capable of focusing the electron beam on the sample such that the sample gets illuminated. As the flow of current increases, the strength of the magnetic lens increases. The electron beam flow is designed such that it cannot pass through the glass lens.

Colorfilters

With hundreds of current applications and countless more waiting to be discovered, multispectral imagery is the key to unlocking insights in any industry.

These excitation fluorescence imaging filters are specifically designed to be used in microscopy and imaging applications. Each Ø25 mm filter is mounted in a 5 mm thick black anodized housing. The housing has an arrow engraved on it that points in the recommended light propagation direction.

DielectricFilters

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Although dichroic mirrors play a crucial role in fluorescence microscopy, they are not perfect when it comes to blocking unwanted light; typically, ~90% of the light at wavelengths below the cutoff wavelength value are reflected and ~90% of the light at wavelengths above this value are transmitted by the dichroic mirror. Hence, some of the excitation light can be transmitted through the dichroic mirror along with the longer wavelength fluorescence emitted by the sample. To prevent this unwanted light from reaching the detection system, an emission filter is used in addition to the dichroic mirror.

A compound microscope is defined as the type of microscope that has more than one lens. It has a combination of lenses and two optical parts known as an objective lens and an eyepiece or ocular lens. The magnifying power of the compound microscope is given as:

Thorlabs' Dichroic Filters are designed to separate light of different wavelengths. When light is incident on the filter at a 45° angle with respect to the normal, the excitation light and its associated back reflection are reflected while the longer wavelength fluorescence signal is transmitted. These filters are unmounted, but are marked by a dash to indicate the coated side of the filter, which light should be incident on. Each filter is 25.0 mm x 36.0 mm. If your application would benefit from a round, mounted dichroic filter, consider our round Dichroic Filters. Click on the icons above to view product-specific transmission data. For further fluorophore compatability, please see the Fluorophores tab.

A simple microscope is defined as the type of microscope that uses a single lens for the magnification of the sample. A simple microscope is a convex lens with a small focal length. The magnifying power of the simple microscope is given as

The working principle of the compound microscope is that the combination of lenses enhances the magnification of the sample. The sample is first viewed as a primary image in the tube and viewed again in the eyepiece.

A single fluorophore can be continually excited unless it is destroyed by photobleaching (i.e. the nonreversible destruction of a fluorophore due to photon-induced chemical damage or covalent modification). The average number of excitation and emission cycles that a particular fluorophore can undergo prior to photobleaching depends on its molecular structure and the local environment; some fluorophores bleach quickly after emitting only a few photons while others are far more robust and can undergo thousands or even millions of cycles before bleaching occurs.

Excitation FilterThe excitation filter only allows a narrow band of wavelengths to pass through it, around the peak fluorophore excitation wavelength. For example, as shown in the graph to the right, the bandpass region corresponding to greater than 90% transmission for the Yellow Fluorescent Protein (YFP) Excitation Filter (MF497-16) is 489 - 505 nm; incident radiation outside of this range is either partially (for regions near the transmission region) or totally (for regions further from the bandpass region) blocked by the filter.

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Lensimaging

The scanning probe microscope has a probe tip that is mounted on the end of a cantilever. The tip is so sharp that it can move precisely and accurately across the surface of the sample scanning every atom. The tip is placed close to the surface of the sample, such that the cantilever experiences a deflection due to forces. This deflection distance is measured by the laser. The final image after scanning is obtained on the computer.

The field of view in a microscope is defined as the diameter of the illuminated circle which is seen through the eyepiece. With an increase in the magnification, there is a decrease in the field of view.

Edgefilters

Each excitation or emission filter is housed in a Ø25 mm black anodized aluminum ring which makes handling easier and enhances the blocking OD by limiting scattering. These filters can be mounted in our extensive line of filter mounts and wheels. As the aluminum rings are not threaded, Ø1" retaining rings will be required to mount the Ø25 mm filters in one of our internally-threaded SM1 lens tubes. For customers who wish to use these filters in Thorlabs, Olympus, or Nikon fluorescence microscopes, Thorlabs manufactures a family of Drop-In Microscope Filter Cubes. Additionally, the unmounted 25 mm x 36 mm dichroic filters can be mounted in the KM2536 kinematic mount, which is designed to secure a 1 mm thick rectangular optic with minimal stress.

The difference in vision between the two eyes is corrected with the help of diopter adjustment. Through diopter adjustment, the focus of the individual eyepiece can be done so that the eyes feel comfortable while observing the sample.

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MidwestFilters

Dichroic MirrorDichroic mirrors are designed to reflect light whose wavelength is below a specific value (i.e. the cutoff wavelength) while permitting all other wavelengths to pass through it unaltered. In a microscope, the dichroic mirror directs the proper wavelength range to the sample as well as to the image plane. The cutoff wavelength value associated with each mirror indicates the wavelength that corresponds to 50% transmission. For example, as shown in the graph to the right, the cutoff wavelength for the Yellow Fluorescent Protein (YFP) Dichroic Mirror (MD515) is ~515 nm. The Specs tab provides information on the reflectance and transmission for each type of dichroic mirror.

These emission fluorescence imaging filters are specifically designed to be used in microscopy and imaging applications. Each Ø25 mm filter is mounted in a 3.5 mm thick black anodized housing. The housing has an arrow engraved on it that points in the recommended light propagation direction.

In this article, there are 5 such microscope types that are discussed along with their diagram, working principle and applications. These five types of microscopes are:

These excitation, emission, and dichroic filters are designed specifically for use in fluorescence imaging applications. They are fabricated at industry-standard dimensions that make them compatible with filter cubes from all major manufacturers. We offer individual filters and filter sets targeted at common fluorophores: BFP, CFP, WGFP, GFP, FITC, Alexa Fluor® 488, YFP, tdTomato, TRITC, Texas Red, mCherry, and Cyanine (CY3.5). In addition, the Fluorophores tab provides information on the alternative fluorophores suitable for these filters. These filters are also available pre-installed into our microscope filter cubes.

By placing one of these mirrors into the experimental setup at 45° with respect to the incident radiation, the excitation radiation (shown in blue in the above right schematic) is reflected off of the surface of the dichroic mirror and directed towards the sample and microscope objective, while the fluorescence emanating from the sample (shown in red in the above right schematic) passes through the mirror to the detection system.

Dichroic filters are marked on the side with the dichroic coating. Light should be incident on this side for best performance.

Click on the below to view the filter set transmission with the absorption and emission spectra of the fluorophore. The key to the right details the meaning of all check marks in the table below. Please note that absorption and emission spectra are unavailable if any is red.

Please Note: The excitation and emission filter housings have an arrow engraved on them that points in the recommended direction of light propagation. Dichroic filters have a marking on the side with the beamsplitter coating. Light should be incident on the marked side of the optic for optimal performance. For further fluorophore compatability, please see the Fluorophores tab.

Long passfilters

Emission FilterAn emission filter serves the purpose of allowing the desirable fluorescence from the sample to reach the detector while blocking unwanted traces of excitation light. Like the excitation filter, this filter only allows a narrow band of wavelengths to pass through it, around the peak fluorophore emission wavelength. For example, as shown in the graph to the right, the bandpass region corresponding to greater than 90% transmission for the Yellow Fluorescent Protein (YFP) Emission Filter (MF535-22) is 524 - 546 nm; incident radiation outside of this range is either partially (for regions near the transmission region) or totally (for regions further from the bandpass region) blocked by the filter.

The fiber optic adapter is used to mate with the fiber optic connector. Many variations are available such as ST, FC, SC and LC.

Absorptivefilters

The depth of focus in a microscope is defined as the distance between the objective lens and the sample plane. The depth of focus varies from person to person and is also dependent on the quality of focus.

Filter DesignOur filters are manufactured to high-performance optical specifications and designed for durability. They are produced via multiple dielectric layers deposited on a high-precision, fused silica substrate. The substrate is ground and polished to ensure that the highest possible image quality is maintained. The resulting hard-coated optics consist of filter layers that are denser than those obtained from electron beam deposition techniques, and which reduce water absorption while greatly enhancing durability, stability, and performance of the filter. Each filter layer is monitored during growth to ensure minimal deviation from design specification thickness, ensuring overall high-quality filter performance.

Filters for Fluorescence MicroscopyThe experimental setup to the right shows the typical filters used for epi-fluorescence microscopy, a form of microscopy in which both the excitation and emission light travel through the microscope objective. By carefully choosing the appropriate filters and mirrors for a given application, the signal-to-noise ratio can be maximized. As shown in the schematic to the right, three types of filters are used to maximize the fluorescence signal while minimizing the unwanted radiation. Each optical element is discussed below.

An electron microscope is defined as the type of microscope in which the source of illumination is the beam of accelerated electrons. It is a special type of microscope with a high resolution of images as the images can be magnified in nanometers.

The working principle of a simple microscope is that when a sample is placed within the focus of the microscope, a virtual, erect and magnified image is obtained at the least distance of distinct vision from the eye that is held at the lens.

The scanning probe microscope is defined as the type of microscope that finds applications in industries where the examination of the specimen is done at the nanoscale levels. The study of a specimen’s properties, its reaction time and its behaviour when stimulated can be done with the help of a scanning probe microscope.

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FluorophoresA fluorophore is a molecule or portion of a molecule that is capable of producing fluorescence. When light of the appropriate frequency necessary to excite a molecule from its ground state to an excited state is present, excitation will occur. However, once in an excited state, the molecule will be unstable. After some short period of time (typically 10-15 to 10-9 s), a photon will be released, thereby enabling the molecule to return to a lower energy state. The emitted radiation will be at a longer wavelength (lower energy) than the absorbed radiation due to the loss of energy through various mechanisms such as vibrations, sound, and thermal energy.

These filters provide excellent transmission of the desired excitation wavelength (>90%), with a sharp spectral cutoff and low transmission at other wavelengths (<0.001%). Click on the icons above to view product-specific transmission data. For additional fluorophore compatability, please see the Fluorophores tab.

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The objective magnification (M) is calculated by dividing the reference focal length (L) of the tube lens by the objective focal length (F). As the critical ...

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A stereo microscope works on the reflected light from the sample. The magnification of the microscope takes place at low power and hence, it is suitable for magnifying opaque objects. It is suitable for thick and solid samples because it uses light reflected from the sample. The magnification of the stereo microscope is between 20x and 50x.

The depth of field in a microscope is defined as the distance from the nearest object plane in focus to the farthest plane in the same focus. In microscopes, the depth of field is very short and is measured in units of microns.

A stereo microscope is defined as a type of microscope that provides a three-dimensional view of a specimen. It is also known as a dissecting microscope. In a stereo microscope, there are separate objective lenses and eyepiece such that there are two separate optical paths for each eye.

There are different types of microscopes and each of these has different purposes of use. Some are suitable for biological applications, while others are used in educational institutions. There are also microscope types that find application in metallurgy and studying three-dimensional samples.

The basic difference between low-powered and high-powered microscopes is that a high power microscope is used for resolving smaller features as the objective lenses have great magnification. However, the depth of focus is greatest for low powered objectives. As the power is switched to higher, the depth of focus reduces.

The table below displays all of the fluorophores that are compatible with our filter sets. The filter set item numbers are listed across the top row and the fluorophores are listed down the first column. Scroll through the table to view fluorophore compatibility with our filter sets.