Once the sample(s) are heat fixed, stain them with crystal violet. This is a purple colored stain, and although both the milk proteins and cells will stain this color, the milk stains faintly and the bacteria will appear dark purple. Keep in mind that the probiotic bacteria are bacilli. Below, sketch a representative field as seen with the oil immersion objective for each of the yogurt samples.

Resolution is often thought of as how clearly the details in the image can be seen. By definition, resolution is the minimum distance between objects needed to be able to see them as two separate entities. It can also be thought of as the size of the smallest object that we can clearly see.

On the flip side, you should be able to see why landscape photographers prefer using f-stops like f/8, f/11, or f/16. If you want your entire photo sharp out to the horizon, this is what you should use.

Once you’ve made the observations using the prepared slide, obtain a glass slide and a sterile swab. Collect a sample from the container of commercially prepared yogurt by swirling the swab in the yogurt, then scraping of the excess on the edge of the container. Smear this over the surface of the slide, making sure that you leave only a thin film of yogurt on the surface. Make a second smear from the container of freshly prepared homemade yogurt, if available. Allow both smears to air dry, and then heat fix them.

A lot of photographers ask me an interesting question: What does the “f” stand for in f-stop, or in the name of aperture (like f/8)?

Light waves that pass through and interact with the object may speed up, slow down, or change direction as they travel through “media” (such as air, water, oil, cytoplasm, etc.) of different densities. For example, light passing through a thicker or denser part of a specimen (such as the nucleus of a cell) may be reflected or refracted (“bend” by changing speed or direction) more than those waves passing through a thinner part. This makes the thicker part appear darker in the image, while the thinner parts are lighter.

The f-stop, which is also known as the f-number, is the ratio of the lens focal length to the diameter of the entrance pupil. If you did not understand that, don’t worry, because there is a much easier explanation of it for beginners. In very simple language, f-stop is the number that your camera shows you when you change the size of the lens aperture.

You might have seen this in your camera before. On your camera’s LCD screen or viewfinder, the f-stop looks like this: f/2.8, f/4, f/5.6, f/8, f/11, and so on. Sometimes, it will be shown without a slash in between like f2.8, or with a capital “F” letter in the front like F2.8, which means the exact same thing as f/2.8. These are just examples of different f-stops, and you might come across much smaller numbers like f/1.2 or much larger ones like f/64.

With a bright-field microscope, images are formed as a result of the interplay between light waves, the object, and lenses. How images of biological objects are formed is actually more physics than biology. Since this isn’t a physics course, it’s more important to know how to create exceptional images of the object than it is to know precisely how those images are formed.

I always find that it’s easiest to understand depth of field by looking at photos, such as the comparison below. In this case, I used a relatively large aperture of f/4 for the photo on the left, and an incredibly small aperture of f/32 for the photo on the right. The differences should be obvious:

Within the past few years, positive health benefits have been correlated with eating fermented foods containing “live” cultures. Although several types of bacteria are known to ferment milk and produce yogurt, two genera in particular, Lactobacillus and Bifidobacterium, have been singled out as promoting good digestive health and a well-balanced immune response. Both of these are bacilli arranged in pairs or short chains. Streptococcus spp., which are cocci arranged in chains, are also usually involved in the process of making the milk into yogurt, but these are not directly associated with positive health benefits to the person who eats the yogurt.

Take a look inside your camera lens. If you shine a light at the proper angle, you’ll see something that looks like this:

Microbiology: A Laboratory Experience Copyright © by Holly Ahern is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License, except where otherwise noted.

Let’s say you wanted to look at cells of Bacillus cereus, which are rod-shaped cells that are about 4 µm long. If you were observing B. cereus with a microscope using the high power lens, how big would the cells appear to be when you look at them? ___________________________

3. When you are finished with the microscope, check the stage to make sure that you don’t leave a slide clipped in the stage. Make sure to switch the microscope OFF before you unplug it. Gently wrap the cord around the base and cover your microscope with its plastic cover.

The other more important impact is depth of field – the amount of your photo that appears to be sharp from front to back. For example, the two illustrations below have different depths of field, depending on the size of aperture:

Usually, the sharpest f-stop on a lens will occur somewhere in the middle of this range — f/4, f/5.6, or f/8. However, sharpness isn’t as important as things like depth of field, so don’t be afraid to set other values when you need them. There’s a reason why your lens has so many possible aperture settings.

It is important to remember that you must use a drop of oil whenever you use the oil immersion objective or you will not achieve maximum resolution with that lens. However, you should never use oil with any of the other objectives, and you should be diligent about wiping off the oil and cleaning all of your lenses each time you use your microscope, because the oil will damage the lenses and gum up other parts of the instrument if it is left in place.

Observe your mouth smear with the microscope. When you get to the oil immersion objective, locate and focus on a single cheek cell. As you did with the prepared slide, sketch the larger cheek cell in the circle provided and label the membrane and nucleus . Add the bacterial cells to your sketch, and try to keep the size scale accurate.

If someone tells you to use a large aperture, they’re recommending an f-stop like f/1.4, f/2, or f/2.8. If someone tells you to use a small aperture, they’re recommending an f-stop like f/8, f/11, or f/16.

As we have previously defined, aperture is basically a hole in your camera’s lens that lets light pass through. It’s not a particularly complicated topic, but it helps to have a good mental concept of aperture blades in the first place.

Locate and focus on a single squamous epithelial cell with obvious bacteria on its surface. Create a sketch of the “cheek” cell (as squamous epithelial cells are sometimes called) in the circle provided. Then label the cell membrane, cytoplasm, and nucleus of the “cheek” cell, which should be easily observed.

These blades form a small hole, almost circular in shape — your aperture. They also can open and close, changing the size of the aperture.

Working f-number

Thank you so much for explaining this topic in a way I finally understand ! I have just started photography using a Nikon d3200 with 70-300mm lens primarily for wildlife .. I wish I had read this article before I started !

This brings us to two additional concepts related to microscopy—working distance and parfocality. Working distance is how much space exists between the objective lens and the specimen on the slide. As you increase the magnification by changing to a higher power lens, the working distance decreases and you will see a much smaller slice of the specimen. Also, once you’ve focused on an object, you should not have to make any major adjustments when you switch lenses, because the lenses on your microscope are designed to be parfocal. This means that something you saw in focus with the low power objective should be nearly in focus when you switch to a high power objective, or vice versa. Thus, for viewing any object and regardless of what lens you will ultimately use to view it, the best practice is to first set the working distance with a lower power lens and adjust it to good focus using the coarse focus knob. From that point on, when you switch objectives, only a small amount of adjustment with the fine focus knob should be necessary.

I'm Spencer Cox, a landscape photographer based in Colorado. I started writing for Photography Life a decade ago, and now I run the website in collaboration with Nasim. I've used nearly every digital camera system under the sun, but for my personal work, I love the slow-paced nature of large format film. You can see more at my personal website and my not-exactly-active Instagram page.

Luckily, you have the building blocks. Aperture and f-stop aren’t complicated topics, but they can seem a bit counterintuitive for photographers who are just starting out. Hopefully, this article clarified some of the confusion, and you now have a better understanding of the fundamentals of aperture.

Move your stage so you observe 10 different microscope fields. Keep track of the number of different types of bacterial cells you encounter during your survey, and record that information below:

F-stop vsaperture

Add to your illustration the bacterial cells which you should see on or near the larger larger cheek cells. Try to keep the size of the bacterial cells to scale with the size of the cheek cell.

2. Clean all of the lenses with either lens paper or Kimwipes (NOT paper towels or nose tissues) BEFORE you use your microscope, AFTER you are done, and before you put it away.

This is a cool concept. It also makes it easy to visualize why an aperture of f/4 would be larger than an aperture of f/16. Physically, at f/4, your aperture blades are open much wider, as shown below:

1. Carry the microscope to your lab table using two hands, and set it down gently on the bench. Once placed on the bench, do not try to slide it around on its base, because this is extremely jarring to the optical system.

The ability of a lens to resolve detail is ultimately limited by diffraction of light waves, and therefore, the practical limit of resolution for most microscopes is about 0.2 µm. Therefore, it would not be practical to try to observe objects smaller than 0.2 µm with a standard optical microscope. In addition, cells of all types of organisms lack contrast because many cellular components refract light to a similar extent. This is especially true of bacteria. To overcome this problem and increase contrast, biological specimens may be stained with selective dyes.

The answer is resolution. Consider what happens when you try to magnify the fine print from a book with a magnifying glass. As you move the lens away from the print, it gets larger, right? But as you keep moving the lens, you notice that while the letters are still getting larger, they are becoming blurry and hard to read. This is referred to as “empty magnification” because the image is larger, but not clear enough to read. Empty magnification occurs when you exceed the resolving power of the lens.

This is why portrait photographers love f-stops like f/1.4, f/2, or f/2.8. They give you a pleasant “shallow focus” effect, where only a thin slice of your subject is sharp (such as your subject’s eyes). You can see how that looks here:

We’ll start by looking at a prepared slide of a “rectal smear,” which is quite literally a smear of feces on a slide stained with a common method called the Gram stain. You will observe several different types of bacterial cells in this smear that will appear either pink or purple. While the main purpose of this is to develop proficiency in use of the oil immersion objective lens, it also provides the opportunity to look at bacteria, observe the differences in cell shapes and sizes, and note that when Gram stained they turn out to be either purple or pink.

By that same logic, an aperture of f/2 is much larger than an aperture of f/16. If you ever read an article online that ignores this simple fact, you’ll be very confused.

Photographers generally don’t care as much about the smallest or “minimum” aperture that the lens allows, which is why manufacturers don’t put that information in the name of the lens. However, if it matters to you, you will always be able to find this specification on the manufacturer’s website. A lens’s smallest aperture is typically something like f/16, f/22, or f/32.

Once the sample is heat fixed, stain it with safranin. This is a pinkish-red colored stain, and all cells (both bacterial and your mouth cells) will take up the stain and increase the contrast in the image.

Based on the picture of the binocular, compound light microscope in Figure 1, match the name of the major part (listed below) with its location on the microscope, and give a very brief description of what each is used for:

One of my friends once had the mirror of his DSLR break in extremely cold weather – not a problem with mirrorless cameras, of course.

Light from an illuminator (light source) below the stage is focused on the object by the condenser lens, which is located just below the stage and adjustable with the condenser adjustment knob. The condenser focuses light through the specimen to match the aperture of the objective lens above, as illustrated in Figure 2.

If you are new to microscopy, you may initially feel challenged as you try to achieve high quality images of your specimens, particularly in the category of “Which lens should I use?” A simple rule is: the smaller the specimen, the higher the magnification. The smallest creatures we observe are bacteria, for which the average size is a few micrometers (μm). Other microscopic organisms such as fungi, algae, and protozoa are larger, and you may only need to use the high power objective to get a good view of these cells; in fact, using the oil immersion objective may provide you with less information because you will only be seeing a part of a cell.

Once you’ve looked at the prepared slide, obtain a glass slide and a sterile swab. Collect a sample of your oral mucosa by gently rubbing the swab over the inside of your cheek. Smear the swab over the surface of the slide (this is known as making a “smear” in microbiology). Allow the smear to dry, and then heat fix by passing the slide through the flame of a Bunsen burner, as demonstrated. Discard the swab in the biohazard waste.

Unfortunately, you can’t just set any f-stop value that you want. At some point, the aperture blades in your lens won’t be able to close any smaller, or they won’t be able to open any wider.

Hopefully, you know how fractions work. 1/2 cup of sugar is much more than 1/16 cup of sugar. A 1/4 pound burger is larger than a 1/10 pound slider.

Why is your aperture written like that? What does something like “f/8” even mean? Actually, this is one of the most important parts about aperture: it’s written as a fraction.

Quite simply, the “f” stands for “focal length”. When you substitute focal length into the fraction, you’re solving for the diameter of the aperture blades in your lens. (Or, more accurately, the diameter that the blades appear to be when you look through the front of the lens).

Aperture f numberphotography

Obtain a prepared slide labeled “yogurt smear” and view it with the microscope. The milk proteins in the yogurt will be visible as lightly stained amorphous blobs. By now you should have a pretty good idea of what bacteria look like, so locate and focus on areas where you see bacterial cells.

Yogurt is produced when lactic acid (homolactic) bacteria that naturally occur in milk ferment the milk sugar lactose and turn it into lactic acid. The lactic acid accumulates and causes the milk proteins to denature (“curdle”) and the liquid milk becomes viscous and semi-solid.

Why is large maximum aperture in a lens so important? Because a lens with a larger maximum aperture lets more light into the camera. For example, a lens with a maximum aperture of f/2.8 lets in twice as much light when compared to a lens with a maximum aperture of f/4.0. This difference could be a big deal when shooting in low-light conditions.

Together we will review how to effectively achieve an exceptional image using a standard optical microscope. This will include not only locating and focusing on the object, but also using the condenser lens and iris diaphragm to achieve a high degree of contrast and clarity.

Adjusting your aperture is one of the best tools you have to capture the right images. You can adjust it by entering your camera’s aperture-priority mode or manual mode, both of which give you free rein to pick whatever aperture you like. That is why I only ever shoot in aperture-priority or manual modes!

You already know the answer to this question, because aperture is a fraction. Clearly, 1/8 is larger than 1/22. So, f/8 is the larger aperture.

Aperture f numbercanon

This is very interesting! As you can see, in the f/4 photo, only a thin slice of the lizard’s head appears sharp. The background of the photo is very blurry. This is known as depth of field.

A lot of photographers really care about the maximum aperture that their lenses offer. Sometimes, they’ll pay hundreds of extra dollars just to buy a lens with a maximum aperture of f/2.8 rather than f/4, or f/1.4 rather than f/1.8.

For example, say that you have an 80-200mm f/2.8 lens fully zoomed out to 80mm. If your f-stop is set to f/4, the diameter of the aperture blades in your lens will look exactly 20 millimeters across (80mm / 4), whereas at f/16, the diameter will be reduced to mere 5 millimeters (80mm / 16).

If you have a 50mm f/1.4 lens, the largest aperture you can use is f/1.4. Professional constant aperture zoom lenses like a 24-70mm f/2.8 will have f/2.8 as their maximum aperture at every focal length. Whereas cheaper consumer-grade lenses such as 18-55mm f/3.5-5.6 will have their maximum aperture change depending on focal length. At 18mm, the maximum is at f/3.5, while at 55mm, it changes to f/5.6. In between is a gradual shift from one to the other.

Here is a final consideration related to objective lenses and magnification. Look at the lenses on your microscope, and note that as the magnification increases, the length of the lens increases and the lens aperture decreases in size. As a result, you will need to adjust your illumination to compensate for a darkening image. There are essentially three ways to vary the brightness; by increasing or decreasing the light intensity (using the on/off knob), by moving the condenser lens closer to or farther from the object using the condenser adjustment knob, and/or by opening/closing the iris diaphragm. Don’t be afraid to experiment to create the best image possible.

That’s a great question. I constantly use my cameras in colder weather than 32F, no matter what they’re rated for. Even sub-zero temperatures (still talking Fahrenheit) don’t concern me, although I don’t think the camera warranty would still apply if something did go wrong in those temperatures.

f-number formula

There would be little to do in a microbiology laboratory without a microscope, because the objects of our attention (bacteria, fungi, and other single celled creatures) are otherwise too small to see. Microscopes are optical instruments that permit us to view the microbial world. Lenses produce the magnified images that allow us to visualize the form and structure of these tiniest of living beings.

As you would expect, there are differences between photos taken with a large aperture versus photos taken with a small aperture. Aperture size has a direct impact on the brightness of a photograph, with larger apertures letting in more light into the camera compared to smaller ones. However, that isn’t the only thing that aperture affects.

Thanks for the explanation. As a newbie I could not figure out why the aperture number increased in value even though the opening decreased. Knowing now that the value is actually a fraction has eliminated the confusion.

Instead, just know that the two biggest reasons to adjust your aperture are to change brightness (exposure) and depth of field. Learn those first. They have the most obvious impact on your images, and you can always read about the more minor effects later.

f-stop photography

Below are some examples of photographs captured at different f-stops from f/2.8 to f/16, to give you an idea of how they are used in the field:

The lens with highest magnifying power is the oil immersion lens, which achieves a total magnification of 1000X with a resolution of 0.2 µm. This lens deserves special attention, because without it our time in lab would be frustrating.

The magnifying power of each lens is engraved on its surface, followed by an “X.” In the table below, find the magnification, and then calculate the total magnification for each of the four lenses on your microscope.

Obtain a prepared slide labeled “mouth smear.” On this slide you will see large cells with a nucleus, clearly visible with both the low power and high power objective lenses. These are squamous epithelial cells that form the outermost layer of the oral mucosa. At high power, you should start to see small cells on the surface of the larger epithelial cells. With the oil immersion objective lens, you will be able to tell the smaller cells are bacteria.

To use this important piece of equipment properly, it is helpful to know how a microscope works. A good place to begin is to learn the name and function of all of the various parts, because when we talk about the ways to improve microscopic images, terms like “ocular lenses” and “condenser” always come up.

When the light waves that have interacted with the specimen are collected by the lenses and eventually get to your eye, the information is processed into dark and light and color, and the object becomes an image that you can see and think more about.

You can think of an aperture of f/8 as the fraction 1/8 (one-eighth). An aperture of f/2 is equivalent to 1/2 (one-half). An aperture of f/16 is 1/16 (one-sixteenth). And so on.

The resolving power of this lens is dependent on “immersing” it in a drop of oil, which prevents the loss of at least some of the image-forming light waves because of refraction. Refraction is a change in the direction of light waves due to an increase or decrease in the wave velocity, which typically occurs at the intersection between substances through which the light waves pass. This is a phenomenon you can see when you put a pencil in a glass of water. The pencil appears to “bend” at an angle where the air and water meet (see Figure 3). These two substances have different refractive indices, which means that light passing through the air reaches your eye before the light passing through the water. This makes the pencil appear “broken.”

Since people care so much about maximum aperture, camera manufacturers decided to include that number in the name of the lens. For example, one of my favorite lenses is the Nikon 20mm f/1.8G. The largest aperture it offers is f/1.8.

The same thing happens as the light passes through the glass slide into the air space between the slide and the lens. The light will be refracted away from the lens aperture. To remedy this, we add a drop of oil to the slide and slip the oil immersion objective into it. Oil and glass have a similar refractive index, and therefore the light bends to a lesser degree and most of it enters the lens aperture to form the image.

F-number calculator

VERY helpful article for this novice. As you are in Colorado, perhaps you can help on this topic: I’m going on a northern lights tour in a few weeks, and am considering getting a camera better than my iPhone. I’ve read many articles recommending various cameras (Sony a6000 keeps coming up), but then when looking at specs, most [affordable in my budget] cameras state an operating temperature only down to 32f. I’ll be in Norway with tens in the teens and 20’s. Are the manufacturer specs to be taken seriously for real-world use? Would I ruin a camera using it in temps 20 degrees cooler than its rating?

Of course, putting everything into practice is another matter. Even if this entire article makes sense for now, you’ll still need to take hundreds of photos in the field, if not thousands, before these concepts become completely intuitive.

You can think of depth of field as a glass window pane that intersects with your subject. Any part of your photo that intersects with the window glass will be sharp. The thickness of the glass changes depending upon your aperture. At something like f/4, the glass is relatively thin. At something like f/32, the glass is very thick. Also, depth of field falls off gradually rather than dropping sharply, so the window glass analogy is definitely a simplification.

For a compound microscope, the optical path leading to a detectable image involves two lenses – the objective lens and the ocular lens. The objective lens magnifies the object and creates a real image, which will appear to be 4, 10, 40, or 100 times larger than the object actually is, depending on the lens used. The ocular lens further magnifies the real image by an additional factor of 10, to produce a vastly larger virtual image of the object when viewed by you.

F-number lens

These are the main aperture “stops,” but most cameras and lenses today let you set some values in between, such as f/1.8 or f/3.5.

The microscope you’ll be using in lab has a compound system of lenses. The objective lens magnifies the object “X” number of times to create the real image, which is then magnified by the ocular lens an additional 10X in the virtual image. Therefore, the total magnification, or how much bigger the object will actually appear to you when you view it, can be determined by multiplying the magnification of the objective lens by 10.

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As a beginner photographer, you might have heard of such terms as f-stop or f-number and wondered what they actually mean. In this article, we will dive into these in detail and talk about how to use them for your photography.

So the microscope makes small cells look big. But why can’t we just use more or different lenses with greater magnifying power until the images we see are really, really big and easier to see?

The second page of our aperture article dives into every single effect of aperture in your photos. It includes things like diffraction, sunstars, lens aberrations, and so on. However, as important as all that is, it’s not what you really need to know – especially at first.

PL provides various digital photography news, reviews, articles, tips, tutorials and guides to photographers of all levels

The human mouth is home to numerous microbes, which persist no matter how many times you brush your teeth and use mouthwash. Since these microbes generally inhabit the surface layers of the oral mucosa, we humans have evolved ways to keep their numbers under control, including producing antibacterial chemicals in saliva and constantly turning over the outer layer of epithelial cells that line the inside of the mouth.

Typically, the “maximum” aperture of a lens, which is also often referred to as “wide-open” aperture, will be something like f/1.4, f/1.8, f/2, f/2.8, f/3.5, f/4, or f/5.6.

That is an important concept! Often, you’ll hear other photographers talking about large versus small apertures. They will tell you to “stop down” (close) or “open up” (widen) the aperture blades for a particular photo.

Appropriate use of the condenser, which on most microscopes includes an iris diaphragm, is essential in the quest for a perfect image. Raising the condenser to a position just below the stage creates a spotlight effect on the specimen, which is critical when higher magnification lenses with small apertures are in use. On the other hand, the condenser should be lowered when using the scanning and low power lenses because the apertures are much larger, and too much light can be blinding. For creating the best possible contrast in the image, the iris diaphragm can be opened to make the image brighter or closed to dim the light. These adjustments are subjective and should suit the preferences of the person viewing the image.