Stereo microscopes work with a sample in real-time and can really be used by anyone who needs to take a closer look at a small object. Veterinarians and their technicians generally use stereo microscopes for dissection or micro-surgery.

Depth of fieldexamples

Skipping one of the 0.00055mm steps that I used means that the step size is approximately 0.0011mm. In the video below you can see that this is (barely) visible. The video shows two alternating stacks. In one of them one single photo is deleted. At this enlargement the area used from one photo is just a thin band of sharp pixels. The difference with and without one photo is a small flickering band of sharp/unsharp.

Depth of field equationcalculator

Some subjects require unique magnification, distinct lighting, and some even require a heated stage. Viewing specimens that require a microscope with particular features can be challenging or even impossible. Knowing roughly what material you will be viewing day to day can aid in your decision about what microscope is going to be the one for your clinic.

Use an eyepiece camera by attaching the small camera to the eyepiece of your microscope. Using a USB port, the eyepiece camera will connect to a computer monitor where you can view and save the images of your microscopic material.

Depth of field equationpdf

Also known as a Charge-Coupled Device Camera, you can easily attach it to a Binocular microscope with a c-mount adapter or a Trinocular microscope with its built-in c-mount adapter. Similar to eyepiece cameras, CCD Cameras will connect to your computer monitor via a USB port. Unlike eyepiece cameras or phone mount cameras, CCD Cameras produce better image quality because of the higher megapixels.

Shallowdepth of field

Vissible light goes from violet approximately 0.380-0.450µm to red approximately 0.625-0.740µm. Green ligth is approximatlelight 0.500-0.565µm. Green light or 0.550µm (0.000550 mm) is a good starting point for the calculations.

Choosing a new microscope for your veterinary practice? You’re likely weighing options between stereo and compound microscopes, while also considering camera compatibility and various other features. Selecting the right microscope is crucial but can be overwhelming. In Intriquip’s guide, we’ll highlight the differences and key factors to consider, helping you make an informed decision for your clinic. Let’s start by understanding what a stereo microscope is and how it differs from its compound counterpart..

When increasing the resolution from 2µm to 1µm the largest step size decreases from 0.028mm to 0.006mm. For an object that is 1mm from top to bottom the NA 0.15 objective needs at least 36 steps/pictures (1/0.028) and the NA 0.30 needs at least 167 (1/0.006) steps/pictures. Using a smaller step size to have some overlap increases the necessary steps/photos even more.

A stereo, stereoscopic, or dissecting microscope is an optical microscope designed for low-magnification observation. Stereo microscopes typically use reflected light that bounces off the surface of an object, rather than light being transmitted through an object.

Depth of fieldphotography

Example calculations of Depth of Field (DOF) using a simplified formula for DOF and using the wavelength (λ) for green light 0.550µm. Case #1: NA=0.14 for example Mitutoyo M plan 5xDOF = 0.550µm / 0,14^2 ≈ 28µm or 0.028mm

The second lens, located in the eyepiece, has a much longer focal point. This allows the image to appear much more enlarged. Compound microscopes are often referred to as biological microscopes, phase contrast microscopes, polarizing microscopes, metallurgical microscopes and fluorescence microscopes. Breaking that down, compound microscopes are used to see a 2D or two-dimensional image.

In practice the step size in focus stacking is usually a bit smaller than the calculated DOF. The reason for this is to have some overlap between the in-focus areas.

Microscope cameras record whatever specimen is being viewed under the microscope in real time. Images or recordings of your microscopic material can be displayed on a digital screen, stored for record-keeping, or shared over the internet in research papers or blog articles. Typically, being used on compound microscopes, there are four common types of microscope cameras.

Another factor to consider is knowing who is going to be using the microscope and what qualifications or training they have. Microscopes can be relatively expensive and require adequate maintenance and cleaning to keep it running smoothly and for a long time. Someone without proper training may not realize this, which can cause damage to an expensive piece of equipment. Providing sufficient training on a new microscope to any and all staff who are going to be using it can help ensure a long lifetime for your microscope.

Once you know what specimens you will be looking at from day to day, you need to consider the different features every microscope offers. No microscopes nor their features are going to be the same. One main feature to look at when purchasing a new microscope is the head. Most commonly, you will find Monocular, Binocular, and Trinocular microscope heads. Another feature to consider is what focus the microscope comes with. Some scopes come with just a coarse focus dial while others have both a coarse and fine focus dial. Lastly, another common feature to consider is the light source.

DOF = Depth of Field in mm λ = Wavelength of the light used in mm. n = Index of refraction NA = Numerical aperture of the objective used

Depth of field equationexample

Unlike stereo microscopes, compound microscopes use light to see in or through a subject which is called light transmission. Most compound microscopes have three or four objectives ranging in magnification from 4x, 10x, 40x, and 100x. Having the second lens in the eyepieces, the total magnification is 40x, 100x, 400x, and 1000x.

DSLR Cameras are also known as Digital SLR Cameras. These cameras require two adapters, a c-mount adapter, and a camera-specific SLR adapter; working best on Trinocular microscopes because of the weight. DSLR cameras have the finest image quality and are a great option when needing to capture images of your subjects.

Depth of fieldphotography examples

Most people have used some form of compound microscope before, usually in a science or biology class in school. Doctors and scientists use compound microscopes to look at bacteria, plant cells, animal cells, chromosomes and even thin layers or slices of organs and tissues. Many veterinarians use compound microscopes in a similar matter typically looking at animal cells, organs, tissues, bacteria and more.

What this really means, is that a stereo microscope shows a 3D or three-dimensional image of whatever microscopic material you are looking at. Commonly, the specimen is visible to the naked eye when using a stereo microscope.

After determining what specimens you are going to be looking at from day to day, you’ll be better equipped in choosing a vet microscope with the necessary features. With the numerous options available, considering who will operate the microscope and their skillset, along with whether a microscope camera is essential, are crucial steps in the purchasing process.. Reach out to Intriquip today to talk more about what microscope is going to best suit your practice.

Numerical aperture (NA) can be used to calculate the Depth of Field (DOF). Calculating DOF is useful for deciding the (largest) step size for focus stacking.

Deepdepth of field

When purchasing a new microscope, there are many things you need to consider to make sure you are getting the right scope for your practice. Knowing what application you need the microscope for is going to be the first crucial thing to consider.

For higher NA and or for other media than air I recommend the formula from the article “Depth-of-Focus in Microscopy” written by I.T. Young, R. Zagers, L.J. van Vliet, J. Mullikin, F. Boddeke, H. Netten. To get the “normal” 2-sided DOF I have multiplied their formula with a factor 2.

Table 1 shows some example calculations of DOF with green light and air. I usually chose a smaller step size than the calculated DOF for green light. Below is an example of two butterfly wing scales that are upside down. This is a focus stack with 255 photos, 0,00055 mm step size, Nikon BD plan apo 40x NA 0.80 microscope objective, Canon 6D camera, stacked in Zerene Stacker.

Microscopes can have LED, Halogen, Tungsten/Incandescent, Fluorescent, or Natural/Sunlight to illuminate a subject. Again, many specimens require different light forms, so knowing what light source is going to work best for those subjects is necessary when purchasing a new microscope.

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Compound microscopes obtain higher levels of magnification and help to diminish chromatic aberration. Compound microscopes use two lenses; one being found in the objectives and one in the eyepieces. Objectives on a compound microscope have a short focal length and are located quite close to the subject, aiding in collecting light to focus on the image.

Simple yet extremely functional, you can easily use your smartphone as a camera for your microscope with the phone mount. Phone mounts usually mount to the top of the scope. You would simply place your smartphone in the mount, so your camera is looking into the eyepiece. You should then see the specimen you looked at through the eyepiece on your smartphone, allowing you to photograph or video it.