Posted on

Top Considerations When Buying a Microscopy Camera, PART 1: Color or Monochrome

Microscopes are found in laboratories and science classrooms around the globe.  With the advent of digital cameras, it is now even easier to upgrade a microscope to view the live image on a screen or monitor, capture images for documentation and analysis, and even stream live images with remote colleagues.

But not all cameras are alike, so how do you know which camera is best for you?  Over the course of this multi-part series, we’ll try to explain the key specifications of cameras and why they matter.  By the end, you should have a good idea of the best camera for your microscope type and application.

PART 1:  Color or Monochrome

Let’s start with an easy question: Do you need a color or monochrome camera?  If you are snapping photos of specimens with many different colors and you need to capture your specimens in all their colorful glory, then a color camera would be best.

But say your specimen is fluorescent, and it doesn’t appear very bright when viewed through the eyepieces.  Even if you may want to view your specimen with different fluorescent channels (different excitation wavelengths and different emission wavelengths, therefore different colors), then a monochrome camera may be your best option.  Let’s explore why. To create a color image, most color cameras use color filters in a distinctive pattern overlayered onto the pixels of the camera sensor.  The Bayer mosaic mask is the most common color filter pattern (see Figure 1).  The tiny filters are arranged in a 2×2 pattern with position 1 being blue, positions 2 and 3 are green, and position 4 is red.  Firmware on the camera itself “fills in the gaps” so that every pixel is interpreted by a computer or monitor as having all three color components – red, blue and green.

Figure 1. Bayer filter pattern (left). Cross-section of sensor and resulting pattern (right). 
Source: https://en.wikipedia.org/wiki/Bayer_filter

By definition, a filter only allows some things to pass through it.  In the case of a color camera, the pixels only “see” a little bit of the light that emanates from the sample.  Monochrome cameras do not have these filters.  For this reason, monochrome cameras are more sensitive than their color counterparts by allowing more light to reach the pixels on the sensor.

Just because monochrome cameras are “color blind” doesn’t mean that you can’t view the image in color.  To make it more appealing to the human eye, most camera software allows the user to apply a color to the monochromatic image, often referred to as pseudocoloring.  Most people will pseudocolor a monochrome image using a color similar to what they would see through the eyepieces (e.g., DAPI has peak emission at 461nm, so a blue of ~461nm is generally used).  In Figure 2, the nuclei were stained with DAPI.  The mitochondria were labeled with a red-emitting fluorophore, and the cytoskeleton was labeled with a green-fluorescing fluorophore.  Each of those channels was pseudocolored according to the emission color of the fluorophore.  Monochrome images, however, allow the user to choose any color to differentiate one stained structure in the sample from another.

Figure 2. Images of fluorescence specimen acquired with monochrome camera. Far left: green excitation, red emission. Second from left: Blue excitation, green emission. Third from left: UV excitation, blue emission. Far right: Composite of each channel after assigning the emission color to the channel.

ACCU-SCOPE is proud to carry microscopy cameras from Teledyne Lumenera.  Lumenera Infinity cameras are available in both color and monochrome versions.  View the selection of Infinity cameras HERE.  ACCU-SCOPE also offers color cameras for a wide variety of applications.  View them HERE.

In the next part of our series, we’ll explore how your microscope configuration may impact your camera selection.

Posted on

Exploring Upright Microscopes: Everything You Need to Know

It is believed that compound microscopes (also known as upright microscopes) have been around since 1590, when Dutch spectacle-maker Zacharias Janssen invented this incredible piece of technology. Since then, scientists and students alike have used upright microscopes to examine specimens that are too small to be visible with the naked eye. What sets them apart from other microscopes is their use of multiple lenses for magnification, flatness correction and color correction. Typical upright microscopes have objective lenses and an eyepiece lens to obtain magnifications between 40x and 1000x. These microscopes are often the go-to choice thanks to their incredible power and simplicity in operation.

To determine if an upright microscope is right for you, let’s look at how it works and in which applications it is best used.

Parts of an Upright Microscope

The components of your particular upright microscope may vary based on the configuration of the instrument, but most uprights have the following features:

  • Eyepiece (ocular): This is the lens through which the observer will look. It typically provides 10x or 15x power.
  • Viewing head: This is the connector piece between the eyepiece and the objective lenses.
  • Arm: This piece connects the head to the base of the microscope.
  • Base: This is the bottom of the microscope.
  • Revolving nosepiece: This part of the microscope holds the objective lenses. Rotate the nosepiece to change to another objective..
  • Objective lens: Typically, an upright microscope will have three or four objective lenses. The shortest lens has the lowest power, while the longest objective usually has the highest power.
  • Stage: This is the platform where the observer places their slides. The stage usually has clips to hold the slides in place. If the stage is mechanical, two knobs on the side will allow the stage to move left and right.
  • Condenser: This component is located under the stage (between the sample and illuminator) and concentrates the light delivery to the specimen.
  • Rack stop: This component determines how close the objective lens can get to the slide. It is typically set in the factory to prevent the lens from coming down on the slide and breaking it.
  • Focus knobs: This component moves the specimen to be in focus through the eyepieces by moving the stage up or down. A coarse focus knob moves the stage faster, while a fine focus knob allows for precise focus of the sample.
  • Illuminator: This is the steady light source shining up through the stage.

How an Upright Microscope Works

Unlike a stereo microscope where the light source usually shines from above onto the specimen, an upright microscope’s light source is transmitted from below the stage and upwards through the sample. The observer then looks down upon the specimen through an ocular lens. The focus knobs adjust the object up or down to be in focus to the eyes. The power of the eyepiece and objective lenses are multiplied together thereby creating greater magnification than a single lens alone. Because the working distance between the objective and the sample is so small, upright microscopes are ideal for thin or flat subjects like bacteria on a slide rather than something that is thicker (e.g., a housefly) or that needs to be dissected.

When to Use an Upright Microscope

Upright microscopes are used to observe small samples or details that are generally too small to be seen with the naked eye. They are ideal for life science and cell biology applications because they can utilize basic observation methods including brightfield, phase contrast, darkfield, and fluorescence microscopy of samples (see examples below).

To reveal greater detail, an upright microscope is often used to view thin slices of larger tissue samples from animals. Brightfield observation uses the inherent color in a sample or samples that are stained to add color to and enhance certain features. Phase contrast uses an optical technique to enhance contrast to usually unstained samples, and some existing color may be lost in this process. In darkfield microscopy, light that strikes a sample is scattered and some is captured by the objective for observation, whereas unscattered light never gets captured, resulting in a dark background. Fluorescence microscopy uses fluorescent molecules to tag particular structures in a sample, making them visible to the eye. Differing from the other observation methods above, the light source for fluorescence microscopy shines from above the sample. Because of their ease of use and powerful magnification, upright microscopes can be readily found in various clinical, educational, pharmaceutical, research, and veterinary laboratories.

Trust ACCU-SCOPE to Meet Your Upright Microscopy Needs

Since its invention over 400 years ago, the upright microscope has been used to make amazing scientific discoveries, and is still the most recognized and popular choice of microscope for viewing specimens on a slide. If you’re shopping for quality microscopes for your lab, turn to ACCU-SCOPE for all your microscopy needs. We have an array of microscopes and accessories, including upright microscopes and microscope heads. If you need low magnification for basic observation and dissection, we also have information on stereo microscopes. Don’t hesitate to contact our team to determine which type of microscope is best for your application.

Posted on

An Exploration into Stereo Microscopes

Stereo microscopes have been in use for a little over a century, but it’s not uncommon for those who routinely use them not to know how they differ from other models. Why are stereo microscopes so special?

With two optical paths and eyepieces, stereo microscopes provide a three-dimensional view of the specimen. Like our eyes, the optical paths converge onto the sample, coming to the focus point from different angles, providing a sense of depth that makes stereo microscopes so beneficial for dissecting applications. They also have a long working distance which allows easy access to the sample with tools. For this reason, stereo microscopes are also referred to as dissecting microscopes.

Let’s take a more in-depth look at stereo microscopes, how they work, and in which applications they should be used.

Parts of a Stereo Microscope

Every component of the stereo microscope is important to its unique function. Its parts may vary depending on the configuration and use of the microscope, but an average classroom stereo microscope will have the following components:

  • Eyepieces: The microscope’s eyepieces are also called ocular lenses and are the part the viewer looks through to examine the specimen. The eyepieces are normally set at 10x magnification, but some models can reach up to 75x magnification.
  • Diopter setting: This microscope component helps prevent eye strain by making up for the differences between the image seen through the left and right eyes.
  • Objective lens: Each eyepiece of the stereo microscope is connected to its own objective lens. The microscope’s magnification level can be determined by a single fixed objective, a rotating multiple lens turret, or a zoom.
  • Stereo head: This component holds the two eyepieces. In one optical design (Greenough), the objective lenses and stereo head are housed together in a single body.  In the second major optical design (Common main objective or Galilean), the head mounts to the top of the optical zoom body.
  • Focus knob: Stereo microscopes are commonly equipped with at least one focus knob. This knob is used to move the stereo head up and down to sharpen the image of the sample.
  • Top lighting: Many microscopes only have one source of lighting, but the stereo microscope has a top and bottom light. The top light illuminates the object from above. Top lighting can be built into the microscope stand, or can be delivered by an external source either attached to the bottom of the microscope (like a ring light) or by fiber-optic bundles that direct light where the user needs it (refer to the feature image for this article).
  • Bottom lighting: The bottom light of the microscope shines light through the stage and improves the visibility of translucent objects.
  • Stage plate: The stage plate is the spot where the specimen is placed. It is directly underneath the objective lens. The plate is often reversible with one black side and one white side to contrast the specimen.
  • Stage clips: These clips are designed to hold any slides in place on the stage.

 

How Does a Stereo Microscope Work?

The stereo microscope is an optical light microscope that uses the light reflected off the specimen to create a clear image of the objects’ individual details. It magnifies things at low power and is commonly used with solid or thick samples.

As the light reflects off the object, the viewer’s left and right eyes see different angles of the same sample, creating a three-dimensional image that is more informative for viewing detail and surface structure than a microscope with a single objective.

Figure 1. Comparison of the two main optical designs of stereo microscopes. Note the separate optical paths for each eye of the observer, and how the optical paths converge onto the specimen at the bottom of the diagram. Also note the longer working distance of the Greenough design (distance from the bottom of the main objective to the specimen).

When Should a Stereo Microscope Be Used?

Because a stereo microscope allows the examination of three-dimensional specimens with its two separate optical paths and long working distance, it can be used in a variety of applications. For example, biologists and students can use these microscopes to perform dissections, while botanists use them to examine plants. They can also aid in repairing circuit boards and watches, cleaning and analyzing fossils, and dermatological examinations.

With additional microscope accessories, the stereo microscope can be configured to match a variety of applications. Boom stands, flex arms, track stands, and table mounts can improve the microscope’s ease of use, and microscope illuminators can enhance the specimen’s image and provide needed contrast.

Looking for Stereo Microscopes? Turn to ACCU-SCOPE

If you’re looking for high-quality stereo microscopes for your research, medical, or instructional purposes, trust ACCU-SCOPE to meet your needs. We have some of the best microscopes in the industry! If you need higher magnifications, we also offer compound microscopes. Contact us today to receive a quote.

Posted on

What to Consider When Investing in Microscopes for Education

Microscopes are wonderful educational tools because they offer students hands-on experiences that bring textbook concepts to life. Because these instruments are vital to a student’s comprehensive learning experience, it’s important to choose the right kind of microscope when outfitting a classroom. A microscope will need to magnify things well, but must also be robust enough to withstand years of use by hundreds of students. Below are four key factors to consider when buying an educational microscope.

Type of Microscope

When choosing a new microscope for the classroom or educational lab, it will be necessary to consider the type of microscope your students will need. What will they be reviewing in class?

If they are looking at larger samples, a stereo zoom microscope could be a good choice for your students. This microscope is often used for biology dissections and examining rocks, minerals, plants, bones, and more. With a stereo microscope, students can get a 3D image of the sample they are studying.

A compound microscope (a.k.a. upright microscope) can be used for samples that the naked eye cannot see because it offers a higher magnification. Samples are prepared and mounted onto microscope slides before being viewed under the lens. Students often look at prepared slides of blood cells, bacteria, tissue, and parasites to save time and complexity of making their own slides.

For live samples such as living cells or organisms, students use inverted microscopes. These microscopes allow them to observe the samples in Petri dishes and other culture vessels. Unlike a compound microscope, the objective lens is located beneath the stage – this offers greater flexibility to accommodate dishes of various sizes.

If you’re still unsure which microscope is suitable for your educational application, you can also look at the size, weight, durability, and ease of use of the microscope to narrow down your choices.

Size

The size of a student microscope is one of the essential features you’ll want to think about when buying a microscope. Most likely, students will be moving the microscopes from storage to their workspace and back every day. In this case, consider a model that is lightweight and compact enough to be transported and stored with ease.

Durability

Often, microscopes in educational environments are used by students having minimal experience with microscopes. For this reason you want to buy a microscope that can withstand bumps, movement, and bounces that are bound to occur. Look for safety features like fixed eyepieces, a locking pin for the observation head, and a focus lock that can help your students’ microscopes to last longer. Microscopes with metal bodies are generally more durable than lighter weight models with plastic features.

Optics

Ensure the microscopes you buy have high-quality optical components so students can visualize all the minor details of a sample. A microscope with a wide magnification range, such as zoom or multiple objectives, enables students to observe samples at multiple magnifications thereby providing context of the detailed structures to the larger specimen. Most newer microscopes offer LED illumination providing energy savings, long lamp life and crisp, even illumination.

Discover the Best Educational Microscope at ACCU-SCOPE

In any science class or student lab, microscopes are essential. With so many types of microscopes available, it can be challenging to narrow down the options while deciding which one to buy. Let the team at ACCU-SCOPE help you discover the best microscopes for your educational purposes. We offer a range of microscope types and accessories, including stereo microscopes, inverted microscopes, and microscope cameras. If you have any questions about our microscopes, reach out to us today!

Posted on

Exploring the Role of Microscopy in Virology and Disease Research

The term microscope originates from the Greek words mikros meaning “small,” and skopein, meaning “to look.” The first recorded use of a microscope-like instrument dates back to the late 16th century. Yet, it wasn’t until the mid-17th century that significant advancement in magnification enabled Van Leeuwenhoek to observe what he called “animalcules” in samples. These “animalcules” were later renamed bacteria and found to be the cause of several diseases such as tuberculosis and the plague. Still, years would pass before viral agents could be successfully observed with microscopy due to past limitations in magnification.

During the latter part of the 19th century, Adolf Mayer speculated that an unknown and unseen infectious agent was causing mosaic disease in tobacco plants. Although he never physically viewed the virus, further experiments involving the filtration of plant particulate matter showed the presence of an undetermined pathogen. It would take decades for the tobacco mosaic virus to be identified in crystallized specimens leading to the additional discovery of over 900 viral variants that infect plant species.

Early Discoveries in Microscopy

In the mid-20th century, the development of the electron microscope enabled researchers to finally observe viruses. By utilizing accelerated electrons, scientists were able to see particles significantly smaller than any elements viewed through the optical microscopes of the time. Although the visual discovery of viral particles was a profound advancement in observational methods, the inability to see viruses work in real-time using an optical microscope would remain elusive until the end of the century.

During the 1980s, light-based observation of viruses was finally possible using highly sensitive microscopes. For the first time, researchers could see viral particles interact with one another and inside cellular membranes. The mechanics of an infection caused by a virus could now be viewed in real-time, enabling scientists to better understand mechanisms of viral infection and illness. The virus could no longer elude detection or the new treatments that would be discovered as a result.

Modern-Day Microscopy

Significant leaps in technological advancements continue in the field of microscopy today. With advances such as phase-contrast microscopy, which can be used to study the impact of viruses on living cells, to TIRF microscopy, which can increase the effectiveness of detection utilizing specialized dyes, the field of microscopy is continuing to open the doors of scientific discovery.

ACCU-SCOPE is a leading provider of microscopes and accessories to provide researchers with robust detection capabilities. The pursuit of knowledge through the scientific process combined with technological advancement paves the way for a brighter future. Through microscopy, we will all benefit from a better understanding of viruses and their impact on us and the environment. To learn more about our microscopes for clinical and research laboratories, please feel free to contact us today.

Posted on

Emerging Trends in Biological Microscopy

As technological forces merge automation and computation with current advancements in optics, the full potential of biological microscopy is coming into focus. Since the advent of phase contrast microscopy by Frits Zernike in 1934, the pursuit of greater illumination techniques has been a driving force behind the many revolutionary advances that have taken place in the field.

The challenge of illumination is ever-present in microscopy. For decades, scientists struggled to find optimal forms of contrast manipulation to better visualize complex details in samples, and to probe deeper into the understanding of life’s complexities. If it were not for the constant pursuit of new techniques and new technologies, the entire scientific and medical communities would still be struggling to find their way in the dark.

2D vs. 3D Imaging

Traditional microscopy typically involves looking at a slice of tissue, cells, or microorganisms sandwiched between two pieces of glass (microscope slide and cover glass). The specimen is so extremely thin — often just a few microns thick — that it seems practically 2-dimensional by our observation. We focus on the specimen, and moving it in X-Y across the stage is generally sufficient to complete our observation.  Information obtained from 2D microscopy in this manner is absolutely beneficial, is the most widely used observation method, and continues to advance scientific knowledge. Yet, cells, tissues, and cellular organisms are 3D entities, and 2D imaging has limitations when exploring the complex mechanisms behind life’s functions. 3D imaging allows scientists and experts to closely examine these processes within a cellular context.

Fluorescence microscopy has truly advanced the application of 3D microscopy for the study of biological processes.  Where 2D microscopy only considers “planar” specimens, fluorescence microscopy adds a third dimension in the form of focus. High-resolution optics and noise-limiting technologies allow the microscope to acquire very thin optical sections through even a single cell (it’s a much more difficult task with thick specimens, but a topic for another time). Stack these images together (referred to as a Z-stack) and a 3D reconstruction of the specimen can be achieved. With the ever-increasing availability of observable data, long-standing mysteries about cellular functions will continue to be solved.

The Transition to Simplified Mechanisms for Non-Expert Users

Recently, there has been a shift in the development of scientific tools so that they can be better utilized by non-expert users. Most microscopes can now be effectively used by individuals with little experience in microscopy. This is due in part to the fact that many scientists and medical professionals are often trained in specialized disciplines. As a result of the depth of specialized training, they generally receive minimal training in microscopy, don’t fully understand how microscopes work (e.g. optics and physics), and don’t know how to optimize the microscope for the best results.

With the advancement of technology, microscopes and many imaging systems (e.g. cameras and software) are now being designed for ease-of-use. The operation of a basic microscope hasn’t changed in over 100 years, but software with a logical interface and retrievable imaging parameters dramatically steepens the learning curve. Microscopes are designed for greater ergonomics, haptics, and visual references (e.g. match the setting on the condenser diaphragm to the Numerical Aperture written on the objective) to simplify operation. It is a careful balancing act between improving technology while retaining accessibility.

Continuing to Shed Light on the Smallest Discoveries

As innovation and discovery continue to advance the field of microscopy, ACCU-SCOPE is here to provide you with the tools to build a solid foundation for your work. We are consistently evaluating our product offerings and adjusting our inventory to feature the latest in microscopy technology. Contact ACCU-SCOPE for more information about cell culture microscopes, clinical microscopes, cameras and digital imaging solutions, and other products that are essential for biological studies.

Posted on

Advantages of Phase Contrast Microscopes for Clinical Settings

Phase contrast microscopes utilize a special technique to enhance the contrast of an image. This technique was developed to produce high-contrast images of transparent or unstained specimens that could not be viewed well under brightfield illumination. Here, we take a look at some of the advantages offered by phase contrast microscopes for clinical settings and other fields that frequently work with living cells, microorganisms, and similar specimens.

Observe Living Cells

Before Dutch physicist Frits Zernike invented the phase contrast microscope in the 1930s, scientists were unable to clearly view living cells. Instead, the organisms were dried on a slide and stained in order produce contrast. However, this process takes time, kills the cell or organism, and can lead to a loss of structure and, therefore, detail. By being able to view an organism while still living, scientists can see how the organism behaves, offering further insight.

View Thin or Transparent Specimens

Specimens that are transparent or translucent, such as very thin slices of tissue, can be difficult to study using brightfield, as much of the detail remains hidden to observation. Phase contrast can make it easier to view and study the details of the internal structure of a cell or microorganism thanks to the increased contrast and minimal manipulation (e.g. fixing) of the specimen.

Can Be Combined with Other Observation Methods

To get the best possible image for their specimen or application, scientists sometimes need to experiment with different observation methods. Phase contrast can be easily combined with other methods of observation, such as fluorescence, to reveal additional detail that couldn’t be observed with just one method.

Applications of Phase Contrast Microscopes

Phase microscopes are not limited to clinical applications. Many different types of specimens can be easily viewed with a phase contrast microscope, including:

  • Live microorganisms, such as bacteria, mold, protozoa, and erythrocytes
  • Fibers
  • Lithographic patterns
  • Glass fragments
  • Subcellular particles, such as nuclei and other organelles

Phase contrast produces a unique “halo” around the edges of particles. This hallmark of phase contrast microscopy is particularly useful to rapidly locate tiny specimens in the sample such as bacteria or debris.

Find Phase Contrast and Other Specialized Microscopes from ACCU-SCOPE

No matter your laboratory’s requirements, we can help you choose the right microscope that fits your needs and budget. ACCU-SCOPE offers microscopes for clinical settings and many other specialized applications. To learn more about our products, or for any other inquiries, please reach out to us online or by phone.

Posted on

How to Choose the Right Microscope for Use in Gemology

Gemologists rely on high-quality microscopes to do their work. Perhaps the most common usage of microscopes in gemology is to conduct an appraisal of a stone. The microscope needs not only to have enough power to be able to see flaws and inclusions in a stone, but also features that accommodate viewing gemstones of all shapes and sizes with ease.

ACCU-SCOPE has an assortment of high-quality binocular and trinocular microscopes for gemologists and jewelry makers that provide the clarity and accuracy demanded of the profession, all at an affordable price point. Here are some features gemologists should consider when shopping for a microscope:

Magnification Power

To see basic details on a stone, 10x power is generally sufficient. However for higher accuracy, many gemologists find that magnification of 40-45x is preferred for viewing the structures inside of a gemstone, including striae, fractures, inclusions, and other elements used to grade gemstones that cannot be seen with the unaided eye or even a loop.

Illumination

In gemology, the most common method for illuminating a gemstone is darkfield illumination, and most gemological microscopes will be equipped with a darkfield base. Patented by Robert Shipley Jr. in 1939, darkfield illumination showcases the gem against a black background. Inclusions and other flaws scatter the light, making them stand out as bright objects against the dark background. Dust and scratches may also scatter light causing them to stand out as well.  Some microscopes also allow the operator to use brightfield illumination, which is useful for examining specimens with different zones of color or to mask dust particles and scratches. Those that photograph gemstones will want both options so that they can represent their subject in the best way possible.

Accessories

Gemstones require special accessories so that they can be easily held and manipulated, observed, and photographed. Depending on your exact needs, you may also want accessories that can hold stones of various sizes as well as jewelry, such as rings. Having a stand that tilts and rotates also makes it easier to view. If you choose a microscope without a base to suit various purposes, you can also purchase a gemological stand.

Find the Right Microscope for You

ACCU-SCOPE has various gemological microscopes along with specialized microscopes for numerous other applications so that you can find the right tool for your needs and budget. Contact us to learn more about our microscopes, get a quote, or find an authorized retailer.

Posted on

Why Should I White Balance?

The short answer to the title question is because white balancing produces better imagesimproves consistencydiscriminates detail, balances contrast, and refines resolution.  If you are using a microscope with a color digital camera, you should definitely take advantage of the white balance feature.  But what is white balance, and when and how should you use it?

What is White Balance?

Quite simply, white balancing is the way we tell a color camera what is white or, at least, neutral (gray).  Camera sensors “see” color according to the color filters that cover the camera sensor – each pixel only “sees” one color: red, green or blue. Depending on the specimen, the pixels will have different intensities and it’s this mixture that recreates the specimen in an image.  When you place a neutral gray or white object on the stage and white balance a camera, the camera makes all the pixels the same intensity – equal amounts of red, green and blue. Now the camera knows what is white or gray, and the colors in the specimen are rendered

Why Does White Balance Change?

The color characteristics of your microscope’s light source varies every time you turn it on.  LED light sources don’t vary as much as halogen lights, but nothing is perfectly stable.  If you have a halogen light, you’ll notice that when the voltage is turned down, the light looks yellowish; turn the voltage up and the light appears white (see Figure 1; used with permission from https://microscopes.unitronusa.com/news/what-is-it-about-white-balancing/).  You will also notice that the color of your sample appears distorted in different color illumination.  White balancing can correct for these variations in light color and normalize your imaging across imaging sessions.  After white balancing the camera, it is important not to adjust the light intensity, especially with a halogen light source – doing so will change the color characteristic of the illumination.  For this reason, it is recommended to 1) adjust image brightness using the exposure control in the camera software, or 2) use neutral density filters to attenuate the light intensity, rather than turning down the voltage (LED light sources may be the exception).

Figure 1.  Impact of Halogen Lamp Voltage on Illumination Color in Microscopy.

When Should I Use White Balance?

You should use white balance at the beginning of every imaging session, after your light source has warmed up, and after you set the light intensity.  If you will be imaging with different magnification objectives, adjust the light intensity for the highest magnification objective then white balance the camera.  As mentioned above, reduce camera exposure time or use neutral density filters to reduce the light intensity for lower magnification objectives – neutral density filters don’t change the color of the illumination.  Alternatively, you can white balance before every image you take.  Whether the images will be used for qualitative or quantitative analysis, white balancing before every image ensures consistency and comparability between them.

How Do I White Balance My Camera?

First, place a specimen on the stage and focus using the highest magnification objective that you plan to use for imaging.  Perform Köhler illumination including condenser iris diaphragm adjustment for the objective being used.

Next, locate the white balance function on your camera or in your camera software.  Most cameras have a function for white balancing, and some may even have a button on the camera housing.  Camera software may have a software button to press for white balancing, or software may have the option for locking white balance that you turn off to white balance, then turn back on to keep that setting. The white balancing process is similar across camera manufacturers.  If your camera has a histogram function, you may find it informative and useful, plus you can see the impact of white balancing on the camera’s color channels (red, green and blue) as well as in the image (see Figure 2).  Adjust the light intensity to about 70%-80% of maximum on the histogram, or approximate a medium brightness if you don’t have a histogram.  Then without changing the light intensity, move the specimen out of the field of view.  Click the white balance function on your camera or in its software.  The background should now be a neutral gray, and the channels on the histogram should overlap.  White balancing the camera does not affect what you see through the eyepieces.

Figure 2. Appearance of imaging field, histogram and specimen before (left panel) and after (right panel) white balancing.

One final recommendation. Automatic white balancing may seem very attractive — your sample will always have the right color. False. The camera adjusts white balance constantly as you move a specimen across the stage or with different samples. Therefore we recommend locking the white balance after you set it.

White balancing is a best practice in microscopy and, as with good experimental design, should be routine in your lab, too.

Thank you for reading.

Posted on Leave a comment

Color Cameras in Microscopy

Believe it or not, people used film for taking pictures with microscopes – big, clunky cameras bolted to the microscopes, often with separate control units, and no way to preview the picture.  Just push the shutter release, cross your fingers and hope for the best – of course you wouldn’t know until you actually developed the film.  Fast forward a few decades and now cameras are ubiquitous – in our pockets, on our mobile phones and devices, even in our doorbells, all ready to capture the moment at a moment’s notice.  However, microscopy cameras are, indeed, different than that ubiquitous smart phone camera.  So why isn’t color imaging in microscopy as simple as a selfie?

Just as with cellphones, there are a wide variety of options for color cameras for microscopes. So how do you know which camera is best for you? Take a closer look at the technology.

Color technology

The most common technology to deliver a color image is a camera featuring a Bayer color mosaic filter (Fig. 1) – a red, green or blue filter on each pixel.  Note that with this most common filter arrangement, a given pixel only “sees” one color — this means that the resolution of a full-color image (in megapixels) is only 25% of the total megapixels available on that camera. A Bayer filter is the same technology used in most professional and consumer cameras, and the cameras in our mobile phones.  It is much more cost effective than 3-chip cameras (different sensor for red, green and blue) or cameras that use liquid filter-based color filters (take sequential images of red, green and blue, then overlay them).  Of course, your application may dictate which technology is most appropriate (i.e. speed, resolution, flexibility, simplicity).

Figure 1. Common Bayer filter pattern on camera color sensor (left panel), and the resulting color pattern recorded by the sensor pixels (right panel.  Images courtesy of https://en.wikipedia.org/wiki/Bayer_filter)

CCD or CMOS?

These acronyms refer to the technology used in the camera sensor.  Traditionally, CCD sensors delivered better quality images (i.e. less noise), and CMOS were cheaper and faster.  Newer CMOS technologies have greatly improved the image quality, and they’re still king when it comes to speed.  Even more recently, sensor manufacturers are concentrating their focus on CMOS technology, and the availability of CCD sensors is declining.

The turtle and the hare

If you are only imaging fixed slides or dead/inanimate objects, then there’s no need for a lightning fast camera – the specimen isn’t going anywhere, literally.  So, go for the best quality image possible, regardless of how fast or slow the camera.  On the other hand, if you are imaging living systems (i.e. live microorganisms) or a specimen in motion, then a faster “shutter speed” translates into better snapshots in time.  For live image streaming, displaying in front of an audience, or video capture, higher speeds are also preferred or necessary (i.e. 30 frames per second or faster is recommended), otherwise the audience could get motion sick.

Cameras with HDMI output generally provide faster frame rates for live preview, and often save the images to an internal storage device.  USB-output cameras used to have slower frame rates than their HDMI siblings, but the newer USB 3.0 cameras can stream at well over 60fps. USB cameras save directly to a PC while offering greater control of camera settings for image acquisition through software.  The latest arrivals are WiFi-connected cameras that take advantage of our mobile devices and a camera app for image acquisition.  Thanks to the latest wireless technologies, their frame rates are usually somewhere between those of HDMI and USB-output cameras.

More is better, right?

As general consumers of imaging technologies (first point-and-shoot cameras, now smart phones), we’re tricked into thinking “more” megapixels translate into better images.  NOT TRUE, at least not for microscopy!  Given the same size of the sensor, more pixels (or the little light-sensing component of a camera sensor) means smaller pixels which, in turn, means less “volume” in the pixel to sense light, thus reducing sensitivity.  Also due to the design of the technologies, there is a little more space between pixels in a CMOS sensor than a CCD sensor, therefore CMOS pixels tend to be smaller than those on a CCD.

And when it comes to sensitivity for lower light applications, [pixel] size does matter.  As I alluded to above, smaller pixels are less sensitive than larger pixels.  So for situations where sensitivity is important (i.e. fluorescence, darkfield, phase contrast), larger pixels (and consequently lower megapixel cameras) are actually preferred.  There is also an ideal pixel size for each microscope magnification, and this is determined based on the resolution of the microscope (please refer to the article “What’s the Deal with Megapixels?”  Suffice it to say that the higher the magnification, the larger the ideal pixel dimensions and, consequently, the fewer the pixels that fit on the sensor.

So, which camera is best?  That’s for you to decide (and our technical applications people can help).  Review the software and, of course, the image quality.  Consider how you will use the images (still images or live viewing), and the camera’s connectivity (HDMI, USB or WiFi).  One last word of advice: take the camera for a test drive.

Thanks for reading!