How microscopes magnify - magnification microscope meaning
As you move through your daily routine, you need to be able to see near, intermediate, and far objects clearly. With multifocal lenses, you’ll be able to clearly see small print on a page, a computer screen, and see objects in the distance.
Each objective is engraved with its class, magnification, numerical aperture, wavelength range, a zero (noting that it is to be used to image a sample without a cover glass), and optical field number. For an explanation of the defining properties of these objectives, please see the Objective Tutorial tab.
Objectivelensmicroscope
The magnification of a system is the multiplicative product of the magnification of each optical element in the system. Optical elements that produce magnification include objectives, camera tubes, and trinocular eyepieces, as shown in the drawing to the right. It is important to note that the magnification quoted in these products' specifications is usually only valid when all optical elements are made by the same manufacturer. If this is not the case, then the magnification of the system can still be calculated, but an effective objective magnification should be calculated first, as described below.
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The effective magnification of the Olympus objective is 22.2X and the trinoculars have 10X eyepieces, so the image at the eyepieces has 22.2X × 10X = 222X magnification.
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The TL1X-SAP, TL2X-SAP and TL10X-2P objectives have parfocal lengths of 95.0 mm, while the TL4X-SAP and TL15X-2P objectives have 60.0 mm and 75.0 mm parfocal lengths, respectively (see the Specs tab for complete specifications). To use these objectives alongside each other, we offer parfocal length extenders; for example, the PLE351 parfocal length extender can be used to increase the parfocal length of the TL4X-SAP objective from 60.0 mm to 95.0 mm.
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The Nikon Plan Fluorite Objectives provide 4X, 10X, 20X, 40X, or 60X magnification. They are designed to have high transmission in the UV to NIR wavelength range and to produce flat images across the field of view. These multi-purpose objectives can be utilized for brightfield microscopy, fluorescence microscopy, and polarization-sensitive observations. The N10X-PF, N20X-PF, N40X-PF, and N60X-PF objectives are also good for both darkfield and DIC microscopy.
An important parameter in many imaging applications is the resolution of the objective. This tutorial describes the different conventions used to define an objective's resolution. Thorlabs provides the theoretical Rayleigh resolution for all of the imaging objectives offered on our site; the other conventions are presented for informational purposes.
Five objective classes are shown in the table to the right; only three common objective classes are defined under the International Organization for Standards ISO 19012-2: Microscopes -- Designation of Microscope Objectives -- Chromatic Correction. Due to the need for better performance, we have added two additional classes that are not defined in the ISO classes.
If you think you need multifocal contact lenses, schedule an appointment with your eye doctor for a comprehensive eye exam and contact lens fitting.
An idealized image of two Airy disks separated by a distance equal to the Rayleigh resolution is shown in the figure to the left below; the illumination source has been assumed to be incoherent. A corresponding horizontal line cut across the intensity maxima is plotted to the right. The vertical dashed lines in the intensity profile show that the maximum of each individual Airy disk overlaps with the neighboring minimum. Between the two maxima, there is a local minimum which appears in the image as a gray region between the two white peaks.
Also referred to as the parfocal distance, this is the length from the shoulder to the top of the specimen (in the case of objectives that are intended to be used without a cover glass) or the top of the cover glass. When working with multiple objectives in a turret, it is helpful if all of the parfocal distances are identical, so little refocusing will be required when switching between objectives. Thorlabs offers parfocal length extenders for instances in which the parfocal length needs to be increased.
Our online and retail stores offer a diverse range of top-quality toric lenses, which come in daily, weekly, or monthly disposables from leading brands.
Thorlabs offers dry objectives made in house, as well as objectives from Olympus, Nikon, and Mitutoyo. Super apochromat, plan achromat, plan apochromat VIS+, plan apochromat, and plan fluorite (also called plan semi-apochromat or plan fluor) designs are available. For details about the differences between these types of objectives, please see the Objective Tutorial tab above.
Each objective is designed for use with a tube lens focal length of 200 mm and is compatible with our DIY Cerna® Systems. They use M25 x 0.75 threading; to use these objectives with a different thread standard, please see our microscope objective thread adapters.
Our selection of dry objectives can be used in applications from microscopy to fiber coupling and includes options optimized for use at wavelengths from the UV to the NIR. For information on recommended applications for specific objectives, see below.
Objectivelens
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Leicaobjective
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The resolution of an objective refers to its ability to distinguish closely-spaced features of an object. This is often theoretically quantified by considering an object that consists of two point sources and asking at what minimum separation can these two point sources be resolved. When a point source is imaged, rather than appearing as a singular bright point, it will appear as a broadened intensity profile due to the effects of diffraction. This profile, known as an Airy disk, consists of an intense central peak with surrounding rings of much lesser intensity. The image produced by two point sources in proximity to one another will therefore consist of two overlapping Airy disk profiles, and the resolution of the objective is therefore determined by the minimum spacing at which the two profiles can be uniquely identified. There is no fundamental criterion for establishing what exactly it means for the two profiles to be resolved and, as such, there are a few criteria that are observed in practice. In microscopic imaging applications, the two most commonly used criteria are the Rayleigh and Abbe criteria. A third criterion, more common in astronomical applications, is the Sparrow criterion.
If you have astigmatism, your contact lens prescription will have additional numbers for Cylinder and Axis. These are specific measurements required for toric lenses that correct astigmatism.
This microscope objective serves only as an example. The features noted above with an asterisk may not be present on all objectives; they may be added, relocated, or removed from objectives based on the part's needs and intended application space.
Objectives can be divided by what medium they are designed to image through. Dry objectives are used in air; whereas dipping and immersion objectives are designed to operate with a fluid between the objective and the front element of the sample.
References[1] Eugene Hecht, "Optics," 4th Ed., Addison-Wesley (2002)[2] S.G. Lipson, H. Lipson, and D.S. Tannhauser, "Optical Physics," 3rd Ed., Cambridge University Press (1995)[3] C.M. Sparrow, "On Spectroscopic Resolving Power," Astrophys. J. 44, 76-87 (1916)
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Objectives following ISO 8578: Microscopes -- Marking of Objectives and Eyepieces will be labeled with an identifier ring to tell the user what immersion fluid the objective is designed to be used with; a list of ring colors can be found in the table to the right.
Example 2: Trinocular MagnificationWhen imaging a sample through trinoculars, the image is magnified by the objective and the eyepieces in the trinoculars. If using a 20X Nikon objective and Nikon trinoculars with 10X eyepieces, then the image at the eyepieces has 20X × 10X = 200X magnification. Note that the image at the eyepieces does not pass through the camera tube, as shown by the drawing to the right.
Their designation as plan achromats indicates that they are flat field and aberration corrected at two different wavelengths in the visible spectrum, leading to better spherical and chromatic corrections and superb field flatness. These achromatic objectives have an ultra-wide antireflection coating and standard RMS threading. To use these objectives with a different thread standard, please see our microscope objective thread adapters.
Example 4: Sample AreaThe dimensions of the camera sensor in Thorlabs' previous-generation 1501M-USB Scientific Camera are 8.98 mm × 6.71 mm. If this camera is used with the Nikon objective and trinoculars from Example 1, which have a system magnification of 15X, then the image area is:
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Using an immersion fluid with a high refractive index allows objectives to achieve numerical apertures greater than 1.0. However, if an immersion objective is used without the fluid present, the image quality will be very low. Objectives following ISO 8578: Microscopes -- Marking of Objectives and Eyepieces will be labeled with an identifier ring to tell the user what immersion fluid the objective is designed to be used with; a list of ring colors can be found in the table above.
All objectives are shipped in an objective case comprised of a lid and container; please see the table to the upper right for compatible replacement cases for each objective. Each objective housing is engraved with the item #, magnification, NA, wavelength range, and working distance. The housings are designed for a tube lens of focal length 200 mm. The TL1X-SAP, TL2X-SAP and TL4X-SAP objectives have M25 x 0.75 external threading, while the TL10X-2P and TL15X-2P objectives have M32 x 0.75 external threading. To use the objectives with a different thread standard, please see our microscope objective thread adapters.
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Objectives with very small working distances may have a retraction stopper incorporated into the tip. This is a spring-loaded section which compresses to limit the force of impact in the event of an unintended collision with the sample.
If you have astigmatism, you may have had difficulty wearing standard contact lenses due to their tendency to move around or become dislodged. Switching to toric lenses may fit your eyes more comfortably and securely, without any blurriness or discomfort.
Multiple optical elements, including the microscope objective, tube lens, and eyepieces, together define the magnification of a system. See the Magnification & FOV tab to learn more.
If an objective is used for water dipping, water immersion, or oil immersion, a second colored ring may be placed beneath the magnification identifier. If the objective is designed to be used with water, this ring will be white. If the objective is designed to be used with oil, this ring will be black. Dry objectives lack this identifier ring entirely. See the table to the right for a complete list of immersion identifiers.
What is the purpose of theobjectivelens in a lightmicroscope
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The images of a mouse kidney below were all acquired using the same objective and the same camera. However, the camera tubes used were different. Read from left to right, they demonstrate that decreasing the camera tube magnification enlarges the field of view at the expense of the size of the details in the image.
The Abbe theory describes image formation as a double process of diffraction [2]. Within this framework, if two features separated by a distance d are to be resolved, at a minimum both the zeroth and first orders of diffraction must be able to pass through the objective's aperture. Since the first order of diffraction appears at the angle: sin(θ1) = λ/d, the minimum object separation, or equivalently the resolution of the objective, is given by d = λ/n*sin(α), where α is the angular semi-aperture of the objective and a factor of n has been inserted to account for the refractive index of the imaging medium. This result overestimates the actual limit by a factor of 2 because both first orders of diffraction are assumed to be accepted by the objective, when in fact only one of the first orders must pass through along with the zeroth order. Dividing the above result by a factor of 2 and using the definition of the numerical aperture (NA = n*sin(α)) gives the famous Abbe resolution limit:
Thorlabs offers a High-Resolution Plan Apochromatic Improved Visible (APO VIS+) Microscope Objective for 400 to 1100 nm which provides axial color correction over a wide field of view with no vignetting over the entire field. Compared to common apochromatic microscope objectives, which are typically axial color corrected from the 436 nm (g-line) to 656 nm (C-line), our PLAN APO VIS+ objective has an extended corrected wavelength range from 436 nm (g-line) to 850 nm. The objective is designed for use with a tube lens focal length of 200 mm and has optical elements that are AR-coated for improved transmission between 400 nm and 1100 nm. For more details on these objectives, please click the info icon () below. Our 50X objective has a high numerical aperture (NA) of 0.75, making it ideal for applications requiring high-resolution such as laser focusing; brightfield, darkfield, and fluorescence microscopy; and two-photon imaging. Thorlabs offers the objective case (Item #s OC2M26 and OC24) separately as a replacement if the case shipped with each of these objectives is lost or broken.
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Objectives following ISO 8578: Microscopes -- Marking of Objectives and Eyepieces will be labeled with an identifier ring to tell the user what immersion fluid the objective is designed to be used with; a list of ring colors can be found in the table to the right.
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The objectives have external W26 x 0.706 threads; to use these objectives with a different thread standard, please see our microscope objective thread adapters. These objectives do not feature adjustment to correct for cover glass thickness and should be used without a cover slip.
The camera sensor dimensions can be obtained from the manufacturer, while the system magnification is the multiplicative product of the objective magnification and the camera tube magnification (see Example 1). If needed, the objective magnification can be adjusted as shown in Example 3.
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Magnification is not a fundamental value: it is a derived value, calculated by assuming a specific tube lens focal length. Each microscope manufacturer has adopted a different focal length for their tube lens, as shown by the table to the right. Hence, when combining optical elements from different manufacturers, it is necessary to calculate an effective magnification for the objective, which is then used to calculate the magnification of the system.
These infinity-corrected, imaging microscope objectives for visible to NIR wavelengths provide 4X, 10X, 20X, 40X, or 60X magnification. Plan Fluorite objectives, also called a plan semi-apochromat, are corrected for four wavelengths. These are well suited for color photomicrography. The RMS60X-PFC features variable coverslip correction; it has a rotating correction collar that changes the distance between the objective elements, allowing the coverslip correction to be adjusted from 0.11 mm to 0.23 mm.
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This objective is engraved with its class, magnification, numerical aperture, a zero (noting that it is to be used to image a sample without a cover glass), and optical field number. For an explanation of the defining properties of this objective, please see the Objective Tutorial tab.
Objectives that feature a built-in iris diaphragm are ideal for darkfield microscopy. The iris diaphragm is designed to be partially closed during darkfield microscopy in order to preserve the darkness of the background. This is absolutely necessary for high numerical aperture (above NA = 1.2) oil immersion objectives when using an oil immersion darkfield condenser. For ordinary brightfield observations, the iris diaphragm should be left fully open.
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What is the job of theobjectivelenses
The term “spherical” simply refers to the shape of the lens, which is round and symmetrical like a sphere, and is used to differentiate these lenses from toric lenses that have a different shape to correct astigmatism.
Following Equation 1 and the table to the right, we calculate the effective magnification of an Olympus objective in a Nikon microscope:
The most common, a standard #1.5 cover glass, is designed to be 0.17 mm thick. Due to variance in the manufacturing process the actual thickness may be different. The correction collar present on select objectives is used to compensate for cover glasses of different thickness by adjusting the relative position of internal optical elements. Note that many objectives do not have a variable cover glass correction, in which case the objectives have no correction collar. For example, an objective could be designed for use with only a #1.5 cover glass. This collar may also be located near the bottom of the objective, instead of the top as shown in the diagram.
Thorlabs provides the theoretical Rayleigh resolution for all of the imaging objectives offered on our site in their individual product presentations.
When imaging a sample with a camera, the dimensions of the sample area are determined by the dimensions of the camera sensor and the system magnification, as shown by Equation 2.
All objectives featured on this page are compatible with our microscope nosepiece modules for DIY Cerna® systems, which accept RMS, M25 x 0.75, or M32 x 0.75 objective threading. Parfocal lengths can be matched by using our parfocal length extenders. The Olympus microscope objectives can be mounted directly to our fiber launch systems, or mounted into our 30 mm cage system using the CP42(/M) RMS-threaded cage plate, which is also post mountable. They can also be mounted to any of our multi-axis platforms or translation stages using an HCS013 RMS mount. Please note that the multi-axis platforms and translation stages need a 3 mm wide central keyway for the HCS013 RMS mount.
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The shoulder is located at the base of the objective threading and marks the beginning of the exposed objective body when it is fully threaded into a nosepiece or other objective mount.
Threading allows an objective to be mounted to a nosepiece or turret. Objectives can have a number of different thread pitches; Thorlabs offers a selection of microscope thread adapters to facilitate mounting objectives in different systems.
To adapt the examples shown here to your own microscope, please use our Magnification and FOV Calculator, which is available for download by clicking on the red button above. Note the calculator is an Excel spreadsheet that uses macros. In order to use the calculator, macros must be enabled. To enable macros, click the "Enable Content" button in the yellow message bar upon opening the file.
For point source separations corresponding to the Rayleigh and Abbe resolution criteria, the combined intensity profile has a local minimum located at the origin between the two maxima. In a sense, this feature is what allows the two point sources to be resolved. That is to say, if the sources' separation is further decreased beyond the Abbe resolution limit, the two individual maxima will merge into one central maximum and resolving the two individual contributions will no longer be possible. The Sparrow criterion posits that the resolution limit is reached when the crossover from a central minimum to a central maximum occurs.
If you visit an eye doctor for an eye exam and find that you have presbyopia, your contact lens prescription will include measurements for Add and Dominant.
Our 1X telecentric objective is ideal for machine vision applications and features a removable magnetic waveplate that minimizes back reflections when used with an epi-illuminated system, thus enabling an increase in contrast; see the image to the right. Our 2X and 4X objectives pair low magnification with NAs of 0.10 and 0.20, respectively, making them ideal for widefield imaging. Lastly, our 10X and 15X objectives are designed for multiphoton imaging applications and provide excellent transmission out to 1300 nm. The 10X and 15X objectives have correction collars that allow adjustment for spherical aberrations introduced by imaging through aqueous solutions or thick cover glasses, without the need for water dipping or oil immersion. The TL15X-2P objective additionally features a locking mechanism to fix the correction collar in place for improved repeatability.
Objectives are commonly divided by their class. An objective's class creates a shorthand for users to know how the objective is corrected for imaging aberrations. There are two types of aberration corrections that are specified by objective class: field curvature and chromatic aberration.
When choosing a microscope objective, it is important to keep in mind that objectives are often designed to integrate with a particular manufacturer's microscopes. Before interchanging objectives, be sure to check the design tube lens focal length and the threading type of the objectives. A full list of specifications for each objective can be found in the Specs tab above. Please note that the performance of each objective may vary from the engraved specifications when integrated with components and systems from different manufacturers. See the Magnification and FOV tab for more information.
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Dipping objectives are designed to correct for the aberrations introduced by the specimen being submerged in an immersion fluid. The tip of the objective is either dipped or entirely submerged into the fluid.
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A cover glass, or coverslip, is a small, thin sheet of glass that can be placed on a wet sample to create a flat surface to image across.
All of these objectives are excellent for brightfield microscopy, while the RMS10X-PF, RMS20X-PF, RMS40-PF, and RMS60X-PFC objectives are also excellent for DIC microscopy. These objectives use standard RMS threading. To use these objectives with a different thread standard, please see our microscope objective thread adapters.
Example 3: Trinocular Magnification (Different Manufacturers)When imaging a sample through trinoculars, the image is magnified by the objective and the eyepieces in the trinoculars. This example will use a 20X Olympus objective and Nikon trinoculars with 10X eyepieces.
Thorlabs offers Mitutoyo Plan Apochromat Objectives with 5X, 7.5X, 10X, 20X, 50X, or 100X magnification. They feature a flat field of focus and chromatic correction over their operating ranges: either 436 nm to 656 nm or 480 nm to 1800 nm. The long working distance provides a wide space between the lens surface and the object making them ideal for machine vision applications. Each objective is engraved with its class, magnification, numerical aperture, a zero (noting that it is to be used to image a sample without a cover glass), and the tube lens focal length for which the specified magnification is valid. For an explanation of the defining properties of these objectives, please see the Objective Tutorial tab. If the case shipped with each of these objectives is lost or broken, Thorlabs offers an objective case (item #s OC2M26 and OC24) that can be used as a replacement.
Example 1: Camera MagnificationWhen imaging a sample with a camera, the image is magnified by the objective and the camera tube. If using a 20X Nikon objective and a 0.75X Nikon camera tube, then the image at the camera has 20X × 0.75X = 15X magnification.
What does theobjectivelens doonamicroscope
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Images can also exhibit chromatic aberrations, where colors originating from one point are not focused to a single point. To strike a balance between an objective's performance and the complexity of its design, some objectives are corrected for these aberrations at a finite number of target wavelengths.
The image to the left below shows two Airy disks separated by the Sparrow resolution limit. As described above, the intensity is constant in the region between the two peaks and there is no intensity dip at the origin. In the line cut to the right, the constant intensity near the origin is confirmed.
Spherical contact lenses, known as single vision contact lenses, are a popular choice that corrects vision for those with nearsightedness (myopia) or farsightedness (hyperopia), as these lenses have the same prescription power throughout the entire lens. They may not be suitable for everyone, especially those with astigmatism and presbyopia.
While toric lenses work great for those with astigmatism, multifocal lenses are ideal for folks who need vision correction for multiple distances, and spherical contact lenses which are the most common for those dealing with near or farsightedness.
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These infinity-corrected, imaging microscope objectives for visible wavelengths provide 4X, 10X, 20X, or 40X magnification. With their high numerical apertures (NA) and large magnifications, they are suitable for focusing or collimating laser light. These Olympus objectives are ideal for imaging applications due to their diffraction-limited performance across the entire visible spectrum. Alternatively, they can be used to focus light to a diffraction-limited spot, enabling efficient coupling of monochromatic or broadband light into a waveguide or fiber. Each of these objectives is suitable for use in brightfield microscopy, while the RMS10X, RMS20X, and RMS40X also offer excellent performance in darkfield imaging.
Thorlabs offers super apochromatic microscope objectives with 1X, 2X, 4X, or 10X magnification, and a plan apochromatic 15X objective with improved visible performance. The objectives are designed to provide axial color correction over a wide field of view with no vignetting over the entire field. Each objective is designed for use with a tube lens focal length of 200 mm and has optical elements that are AR-coated for improved transmission. For more details on these objectives, please click the info icons () below or see the full presentation.
In order to facilitate fast identification, nearly all microscope objectives have a colored ring that circumscribes the body. A breakdown of what magnification each color signifies is given in the table below.
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As with any type of contact lenses, the best contacts for nearsightedness and farsightedness depend on your vision needs, lifestyle, and personal preferences. Consult an eye doctor to find the most suitable option for you.
To get spherical contact lenses, schedule an eye exam with an eye care professional. Your eye doctor will determine if they are right for you and give you a contact lens prescription. Use this to purchase your lenses and follow your doctor’s instructions for proper wear and care.
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Lumenmicroscope
These lenses are specifically designed with the proper shape and prescription power to fit the eye better and provide clear vision. They provide much more effective and comfortable vision correction for people with astigmatism compared to “regular” (spherical) contacts.
They will give you a contact lens prescription and help you choose the best type of toric lenses for you. During a contact lens fitting appointment, your doctor will ensure that the lenses fit properly and show you how to insert, remove, and care for your lenses.
Thorlabs MicroSpot objectives provide long working distances while keeping axial focal shift low. Their optical design is chromatically optimized in the UV wavelength range. Diffraction-limited performance is guaranteed over the entire clear aperture. These objectives are ideal for laser cutting, surgical laser focusing, and spectrometry applications. They can also be used for scanning and micro-imaging applications like brightfield imaging under narrowband, UV laser illumination. Each objective is shipped in an objective case comprised of an OC2M26 lid and an OC24 canister.
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Bifocal lenses have only two prescriptions, one for distance and another for near vision, while multifocal lenses have a range of prescriptions throughout the lens.
As the magnification increases, the resolution improves, but the field of view also decreases. The dependence of the field of view on magnification is shown in the schematic to the right.
The working distance, often abbreviated WD, is the distance between the front element of the objective and the top of the specimen (in the case of objectives that are intended to be used without a cover glass) or top of the cover glass. The cover glass thickness specification engraved on the objective designates whether a cover glass should be used.
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These objectives are designed for use from -18 °C (0 °F) to 60 °C (140 °F) and are not recommended for use at extreme temperatures.
They will be able to determine if multifocal contact lenses are right for you and prescribe the appropriate type of lens on your contact lens prescription, which will be valid for up to 2 years.
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The labeling area for an objective usually falls in the middle of the objective body. The labeling found here is dictated by ISO 8578: Microscopes -- Marking of Objectives and Eyepieces, but not all manufacturers adhere strictly to this standard. Generally, one can expect to find the following information in this area:
Immersion objectives are similar to water-dipping objectives; however, in this case the sample is under a cover glass. A drop of fluid is then added to the top of the cover glass, and the tip of the objective is brought into contact with the fluid. Often, immersion objectives feature a correction collar to adjust for cover glasses with different thicknesses. Immersion fluids include water, oil (such as MOIL-30), and glycerol.
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Additionally, the objective label area may include the objective's specified wavelength range, specialty features or design properties, and more. The exact location and size of each and any of these elements can vary.
At the Sparrow resolution limit, the center of the combined intensity profile is flat, which implies that the derivative with respect to position is zero at the origin. However, this first derivative at the origin is always zero, given that it is either a local minimum or maximum of the combined intensity profile (strictly speaking, this is only the case if the sources have equal intensities). Consider then, that because the Sparrow resolution limit occurs when the origin's intensity changes from a local minimum to a maximum, that the second derivative must be changing sign from positive to negative. The Sparrow criterion is thus a condition that is imposed upon the second derivative, namely that the resolution limit occurs when the second derivative is zero [3]. Applying this condition to the combined intensity profile of two Airy disks leads to the Sparrow resolution:
Note that Leica, Mitutoyo, Nikon, and Thorlabs use the same tube lens focal length; if combining elements from any of these manufacturers, no conversion is needed. Once the effective objective magnification is calculated, the magnification of the system can be calculated as before.
Thorlabs can provide these objectives with custom AR coatings on request by contacting Tech Support; options include broadband NUV (325 nm - 500 nm), dual band (266 and 532 nm), and laser line (248 nm, 266 nm, 355 nm, or 532 nm). We also offer additional MicroSpot objectives for laser-focusing applications in the UV as well as visible and near-IR wavelengths.
In the image below, two Airy disks are shown separated by the Abbe resolution limit. Compared to the Rayleigh limit, the decrease in intensity at the origin is much harder to discern. The horizontal line cut to the right shows that the intensity decreases by only ≈2%.
Field curvature (or Petzval curvature) describes the case where an objective's plane of focus is a curved spherical surface. This aberration makes widefield imaging or laser scanning difficult, as the corners of an image will fall out of focus when focusing on the center. If an objective's class begins with "Plan", it will be corrected to have a flat plane of focus.
The Rayleigh criterion states that two overlapping Airy disk profiles are resolved when the first intensity minimum of one profile coincides with the intensity maximum of the other profile [1]. It can be shown that the first intensity minimum occurs at a radius of 1.22λf/D from the central maximum, where λ is the wavelength of the light, f is the focal length of the objective, and D is the entrance pupil diameter. Thus, in terms of the numerical aperture (NA = 0.5*D/f), the Rayleigh resolution is:
Here, the Design Magnification is the magnification printed on the objective, fTube Lens in Microscope is the focal length of the tube lens in the microscope you are using, and fDesign Tube Lens of Objective is the tube lens focal length that the objective manufacturer used to calculate the Design Magnification. These focal lengths are given by the table to the right.