The World of Photomicrography, As Told By Michael Peres and Nathan Renfro

The World of Photomicrography, As Told By Michael Peres and Nathan Renfro

 There are many reasons why no two snowflakes are alike. All snowflakes start out as a six-sided structure called a plate. As the plate grows, wings form and the ice crystal then becomes a dendrite. Not all storms provide snowflakes that look like this. Sometimes the snow is granular and appears almost like chipped ice. Snowflakes can be columns, needles, or can also exhibit a frost-like condition called rime. © Michael Peres

There are many reasons why no two snowflakes are alike. All snowflakes start out as a six-sided structure called a plate. As the plate grows, wings form and the ice crystal then becomes a dendrite. Not all storms provide snowflakes that look like this. Sometimes the snow is granular and appears almost like chipped ice. Snowflakes can be columns, needles, or can also exhibit a frost-like condition called rime. © Michael Peres

"I get to discover new alien worlds filled with some of the most amazing examples of natural beauty. Having a camera on my microscope just lets me bring others along the journey....and I get to do this without having to get on a plane, drive hours in a car, hike lots of miles. I just get to sit in my office and go on an expedition whenever I want, for as long as I want.”

Michael Peres

By Hannah Loesch

Michael Peres

Michael Peres is an award-winning photo educator and science photographer, a professor of biomedical photography, and and teacher of photomicrography, biomedical photography and other related applications of photography in science at Rochester Institute of Technology. Since 1973, Peres has enjoyed a varied photographic career, and has been actively publishing most of his career. He is the author of Laboratory Imaging and Photography, published by Focal Press in 2016, and co-author of Michael Photographs a Snowflake, a children’s book published by Fossil Press also in 2016. Michael holds a master's degree in instructional technology and bachelor's degrees in biology and biomedical photographic communications. He is also a registered biological photographer who is best known for, in his words, photographing “very tiny things” using a microscope.

 This photograph reveals the various types of cells found in a typical complex and woody plant stem. Visible are vascular bundles and many other specialized cells. The photograph was made using brightfield illumination. The object was 5mm in diameter. © Michael Peres

This photograph reveals the various types of cells found in a typical complex and woody plant stem. Visible are vascular bundles and many other specialized cells. The photograph was made using brightfield illumination. The object was 5mm in diameter. © Michael Peres

Hannah Loesch: Obviously, you’re no stranger to science or photography. Do you need a science background to photograph microscopic things?                                                                          

Michael Peres: I serve as the Associate Chair of the century-old RIT School of Photographic Arts and Sciences, and I am a professor of biomedical photographic communications. I joined the RIT faculty in 1986. I teach courses that explore biomedical photography, and in particular, light microscopy. Having a science background when photographing scientific objects has been very helpful. Being aware of the “science” of the subject and then subscribing to scientific methods is a fundamental expectation in the broadly-defined field of science. I believe having a core knowledge about the subject has been an important tool for my work in the same way a creative who possess an art background finds it useful for creating new work that might build on successful approaches of those who have come before. Knowledge can influence an appropriate way forward. Science photographs are typically produced for scientific consumption. For many in the field, the photograph is made to serve as a permanent record of a subject that records and preserves data about the subject or a state of its being when in a dynamic flux and changing over time. I have been fortunate to have earned two degrees. One degree is in pre-medical studies and the other is in biomedical photographic communications. This foundational knowledge has provided opportunities and access to research and imaging problems beyond my wildest imagination.

 This picture features a wide field high-resolution image of fetal cat skull. It was prepared for examination and cut in a longitudinal view. The image is result of four individual images that were combined into one final file called a computational photograph. Files made this way have more resolution and can be more greatly enlarged. The photograph reveals the maturing ears, eyes, skull bone, sinus cavities, tongue and various other anatomical features of the skull. The sample was 12mm in diameter. © Michael Peres

This picture features a wide field high-resolution image of fetal cat skull. It was prepared for examination and cut in a longitudinal view. The image is result of four individual images that were combined into one final file called a computational photograph. Files made this way have more resolution and can be more greatly enlarged. The photograph reveals the maturing ears, eyes, skull bone, sinus cavities, tongue and various other anatomical features of the skull. The sample was 12mm in diameter. © Michael Peres

HL: What would you call yourself, officially? Is photomicrographer correct?

MP: First and foremost I guess I am a photo educator, but yes, the making of pictures using a microscope is called photomicrography. And so when I am photographing at a microscope, I am sometimes a photomicrographer. Many trained and untrained science photographers refer to this process as photomicroscopy. There is no such thing. “Scopy” comes from the Greek word used to describe viewing and “graphy” describes writing.  Like many professionals, I wear many hats and use many tools. I use close-up lenses, stereo photomicroscopes, simple microscopes, and compound photomicroscopes. Depending on whether the subject is transparent or opaque, there are also decisions about what instrument and illumination are best to reveal the structure or behaviors of a microscopic subject.

 This picture features a singular diatom. Diatoms are microalgae that are also phytoplankton. Diatoms are singular cellular organisms. They have a silica shells called a frustule. This picture has an approximate magnification of 400x. © Michael Peres

This picture features a singular diatom. Diatoms are microalgae that are also phytoplankton. Diatoms are singular cellular organisms. They have a silica shells called a frustule. This picture has an approximate magnification of 400x. © Michael Peres

HL: Are you a scientist first, or a photographer first?                                                                

MP: When preparing to photograph, I am a scientist first. Once the object is prepared and ready for photography, I locate it under an objective (lens), and I then begin thinking about the same things other photographers wrestle with when making pictures. Composition, isolation, focus, sharpness, effective lighting, finding a strong focal point and a myriad of other less important things. I first have to make the subject visible, and then I can make a photograph. If I can’t see it, I cannot photograph it. Science photographers have a unique set of problems to resolve. Knowing as much about the object is a huge advantage for me. Science photographs first and foremost capture and reveal the science of the sample.

 This picture features a garden snake tongue revealed in longitudinal section. Visible are various intrinsic muscle groups which allows the tongue’s shape to be controlled for both smell, touch and taste. Also evident is the outer layer of epithelial or skin cells that surround the tongue which was stained during its sectioning by a trained histologist.  The image is result of four individual images that were combined into one final file. The tongue was 4mm in diameter. © Michael Peres

This picture features a garden snake tongue revealed in longitudinal section. Visible are various intrinsic muscle groups which allows the tongue’s shape to be controlled for both smell, touch and taste. Also evident is the outer layer of epithelial or skin cells that surround the tongue which was stained during its sectioning by a trained histologist.  The image is result of four individual images that were combined into one final file. The tongue was 4mm in diameter. © Michael Peres

HL: Can you describe your process for capturing images?                                                      

MP: Prior to ever making a photograph, it is a common practice to research the subject and identify clear objectives as to why the photographs are being produced. I also want to know what I should be looking for and what I might or might not see. Once informed, and depending on the subject, I must prepare an object for examination and imaging. I use razor blades, very fine paint brushes, distilled water baths, or I sometimes purchase prepared samples from scientific supply companies such as Wards Natural Science or Carolina Biological to get the subjects I am seeking. Preparing to photograph microscopic things also includes boring activities such as cleaning my lab bench and the surface of the lenses I will use. I need to select the proper illuminators, formatting memory cards and other not so interesting minutia. And when photographing #tinythings, my success resides in paying attention to the smallest of things. I photograph more dirt and dust sometimes than subjects.

 This picture features the seeds of Taraxacum officinale, or common dandelion. Dandelions are herbaceous perennial plants from the Asteraceae family. This picture features the mature seeds in a seed pod ready dispersal. The pod has been sectioned in the longitudinal direction revealing the fertilized seeds within a seed coat, a sort of armor there to protect the seed. An individual flower can produce between 40-100 seeds. The seeds are attached to the sepal which opens to become the seed cup. This sample was 6mm in length and was magnified 10 times by the microscope. The tissue was stained with a special green and red dye useful to separate the different types of cells that comprise the preparation. © Michael Peres

This picture features the seeds of Taraxacum officinale, or common dandelion. Dandelions are herbaceous perennial plants from the Asteraceae family. This picture features the mature seeds in a seed pod ready dispersal. The pod has been sectioned in the longitudinal direction revealing the fertilized seeds within a seed coat, a sort of armor there to protect the seed. An individual flower can produce between 40-100 seeds. The seeds are attached to the sepal which opens to become the seed cup. This sample was 6mm in length and was magnified 10 times by the microscope. The tissue was stained with a special green and red dye useful to separate the different types of cells that comprise the preparation. © Michael Peres

Like any photographer, I have to prepare my equipment. Unlike other photographers of other genres who photograph large things, my subjects can often be invisible to the unaided eye. There are no typical things that I might examine, and each is unique in many ways. Each subject comes with its own set of problems. Many objects are too large to examine under a microscope. If this is the case, I must prepare the subject of microscopic examination by dissecting some material from the larger sample. X-ACTO Knives, razor blades, tweezers, insect pins and ping-pong balls are elements in my tool box.

 This picture features muscovite biotite granite, a North American mineral. The photomicrograph was made using polarized light. This reveals the birefringence (multiple refractive indices), and is evidenced by the appearance of the various colors in the picture. The picture is approximately 100x. © Michael Peres

This picture features muscovite biotite granite, a North American mineral. The photomicrograph was made using polarized light. This reveals the birefringence (multiple refractive indices), and is evidenced by the appearance of the various colors in the picture. The picture is approximately 100x. © Michael Peres

Once the object is under my lens, focusing, isolating, and making the lighting for the object is the next challenge. Everything has to be miniaturized. Making images that are larger than the subject from which they came produces some specific problems to the zone of focus, often called depth of field. When magnifying something 10x, it is not uncommon to have a zone of focus of approximately 1mm. This represents the distance between your fingers when just letting the light creep between the crack. Some subjects require the use of computational photography, where I extend the zone of focus using image processing software that builds new images from image slices.

 This photomicrograph reveals the various tissues contained within an undiagnosed skin biopsy. Evident in the picture are the various layers of skin including connective tissues and the possible basal cell carcinoma. Peres made the picture as a B&W file because he felt the natural design of tissue’s organization become more dramatic with absence or color. The magnification is approximately 100x. © Michael Peres

This photomicrograph reveals the various tissues contained within an undiagnosed skin biopsy. Evident in the picture are the various layers of skin including connective tissues and the possible basal cell carcinoma. Peres made the picture as a B&W file because he felt the natural design of tissue’s organization become more dramatic with absence or color. The magnification is approximately 100x. © Michael Peres

Even when I make really lovely photographs, the results might not be correct. I was recently photographing a 500-million-year-old fossil of a sponge collected in Decatur, Georgia. I was provided highly detailed notes about the subject, but because I am not a paleontologist, I did not photograph the proper structures that were most important to the scientist. The photographs were technically perfect, but the science was not properly revealed.

HL: You are known for capturing photos of snowflakes. How do you get an image of something so fragile, from just the right angle, before it melts?                                        

 This picture features a stellar plate crystal of a snowflake photographed March 13, 2013. The crystal was approximately 2mm. All snowflakes are different in appearance for many reasons. This crystal has not fully developed the dendritic wings that are often expected in snowflakes. © Michael Peres

This picture features a stellar plate crystal of a snowflake photographed March 13, 2013. The crystal was approximately 2mm. All snowflakes are different in appearance for many reasons. This crystal has not fully developed the dendritic wings that are often expected in snowflakes. © Michael Peres

MP: Photographing snowflakes presents special problems for sure. Keeping the crystal frozen is one problem of many. I have cultivated simple approaches that lead to predictable outcomes. When the right type of snowflakes are falling, which surprisingly is not that many times, I catch the crystals on a piece of black velvet. Annual snowfalls in Rochester, N.Y. can exceed 100+ inches, but beautiful, intricate dendritic crystals compromise a mere fraction of that accumulation. Mostly the crystals I find appear like chipped ice or Styrofoam. I am sure it is apparent, but it should be stated that I have to work outside and all of my equipment must be kept below freezing. Because of my own situation at my home snowflake shack, I cannot photograph unless the air temperatures are below 30°F.

 © Michael Peres

© Michael Peres

Once I have caught some possible subjects in my velvet-lined catch tray, I carefully but quickly scan the bounty for the subject of my next best snowflake photomicrograph. Using a needle taped to a pencil, I pick up the crystal and transfer it to a glass slide. I locate the slide under my homemade snowflake microscope built from old and discarded pieces and parts from instruments long retired from previous work.

 © Michael Peres

© Michael Peres

Once placed onto the instrument’s aerial platform, I focus the image and work with fiber optic lights to produce both brightness and darkness within the crystal. Snowflakes, like gemstones, have a number of faceted surfaces and unique topography. Each crystal’s internal and external appearance is one of kind because of the process that led to its formation. I can selectively feature elements of the crystal through this method. Individual characteristics on a flake are influenced by the length of time aloft, temperatures both at formation and those the crystal experiences during its growth, static electricity, humidity and wind conditions. Each of these play a role in the ultimate shape and size of a crystal.

 © Michael Peres

© Michael Peres

Depending on air temperatures, I may only have seconds before the crystal begins to change. These changes are evidenced as melting or sublimation, a phenomena where the crystal evaporates and shrinks rather than melting. If the air temperatures are below 25°F, I can work with a crystal for possibly a minute. After that, I have to move onto a new one. The bands of snow are frequently changing during a storm, which can be maddening, and I might be finished before I even start.

 This picture features a stellar plate crystal of a snowflake photographed December 14, 2013. The crystal was approximately 2mm. © Michael Peres

This picture features a stellar plate crystal of a snowflake photographed December 14, 2013. The crystal was approximately 2mm. © Michael Peres

Focusing is a challenge because everything is changing. Even my own vision changes. When studying these tiny ice structures in my viewfinder, my vision gets fatigued. Mostly I am in the zone when photographing and do not feel the cold initially, until my fingers and toes are numb.

It is a pretty interesting experience to photograph things that exist for only moments, and that you are only person to have seen. The photograph is the only marker of its existence. The geometry, the natural design, the uniqueness of each subject, and the rush I get each and every winter is exhilarating. By March though, I hate winter and long for summer and warmer temperatures.

 Michael Peres catching snowflakes on a black velvet catch tray. © Michael Peres

Michael Peres catching snowflakes on a black velvet catch tray. © Michael Peres

HL: You’ve photographed crystals visible within the mineral biotite granite. In an Instagram caption for one of these photos, you pointed out different sections of color within the crystal, and said that each color represents an inclusion. What does that mean? Are these colors altered after the photo is taken to better identify different parts of the object, or is that exactly what you saw through the microscope?                                                                                   

 © Michael Peres

© Michael Peres

MP: Geological and a few other types of samples exhibit what is called birefringence. Birefringence can be observed by the presence of rainbow-like colors within a sample when examined using polarized light. The ability to assess or identify what mineral is present in a sample is a highly specialized and technical skill. It is used in the geological and forensic sciences with great frequency. When I photograph mineral samples, it is mostly to share the intrinsic beauty, shape, structures and natural design. When I set up the microscope, I can move a number of optical components to enhance or diminish the presence and saturation of the colors produced from a sample. Very few minerals are pure and most samples have a variety of elements within the sample. The colors produced from the sample and the polarized light are truthful indicators of the sample’s composition, and are not altered or exaggerated. The colors that have been recorded are fairly consistent with what is observed in the eye pieces.  

 This photomicrograph features the flower of Taraxacum officinale, or common dandelion. It was sectioned and stained for microscopical examination by a botanist. The flower cup holds the now ready for disbursal mature seeds held within. The flower has a diameter of 8mm. © Michael Peres

This photomicrograph features the flower of Taraxacum officinale, or common dandelion. It was sectioned and stained for microscopical examination by a botanist. The flower cup holds the now ready for disbursal mature seeds held within. The flower has a diameter of 8mm. © Michael Peres

HL: What do you think is so fascinating about seeing things at a micro level?     

MP: I keep an open mind about what I expect to see or not see, and I am always surprised. Sometimes in personal explorations with new materials, the sessions are inspired by curiosity and pursuing photographs that possess truthfulness about objects. That being shared, I try to photograph in new ways. I think almost everything has been photographed once in this world, so I pursue new ways of exploring a subject. I keep my eyes and mind open to possibilities while subscribing to tried and true approaches that are industry accepted and not too big of a departure from acceptability. My curiosity for this work was spawned in 1976 in a histology class. From my first extended work at microscope, I found myself repeatedly drawn back to see more of what you cannot see without a microscope. Now 40 years later, I am still being surprised at what I can see and the images I make. Besides my beloved snowflakes, I particularly love botany and biology, too.

 This picture features a cross section of a pine stem. It was photographed using a low power objective. Four pictures were made of the sample and stitched into a composite file shared here. The sample was illuminated using darkfield illumination to make the image more dramatic. Visible in the photomicrograph is the old wood in the middle (sometimes called pith); the growing wood outside of the pith; the xylem, responsible for water transport; and the phloem, the outermost layer that transports the simple sugar produced from photosynthesis. The outermost layer is called epidermis or bark. The stem was 5 mm in diameter and the microscope produced a magnification of 25x. © Michael Peres

This picture features a cross section of a pine stem. It was photographed using a low power objective. Four pictures were made of the sample and stitched into a composite file shared here. The sample was illuminated using darkfield illumination to make the image more dramatic. Visible in the photomicrograph is the old wood in the middle (sometimes called pith); the growing wood outside of the pith; the xylem, responsible for water transport; and the phloem, the outermost layer that transports the simple sugar produced from photosynthesis. The outermost layer is called epidermis or bark. The stem was 5 mm in diameter and the microscope produced a magnification of 25x. © Michael Peres

HL: Do you have a specific goal in mind when you shoot these photos? Are they simply meant to be visually appealing, or do you use them for scientific purposes as well?                                                                                                                                          

MP: If I am photographing for a person or an organization, the objectives for the photographs are clearly defined and I subscribe to their accepted practices and expectations. My approaches must produce images that are highly accurate to the object’s appearance and cannot embellish features or behaviors. My pictures create science facts and are not science fiction. My pictures exhibit an interesting duality in that they can function in science and sometimes in a gallery. They are precise photographic renditions of a subject produced using the best practices, but they often result in intrigue and curiosity about the object because of their presentation as a two-dimensional photograph.

 The stem of a buttercup, shown in cross section. © Michael Peres

The stem of a buttercup, shown in cross section. © Michael Peres

Unique and visually-interesting structures or organization of tissues leads to this visual appeal to a lay audience, I believe. I also love to share science pictures that surprise viewers. I once photographed a basal cell carcinoma tissue biopsy. I made circular photographs and composed them in such a way as there was a strong horizon line. Viewers were captivated by the shapes and layout of the cell structures and once informed it was skin cancer, were horrified. I still maintain that photographs of the invisible and of science are not mainstream, even though social media has broadened the distribution of science photography. My science photographs and those of many of my contemporaries can operate in both arenas. Maybe in the end analysis, they are both and no singular definition fits. Maybe my intent would play a more central role in answering the question about how they function, but so does the perspective of the viewer.

Nathan Renfro

 Polarized light shows vibrant birefringent colors from optically misaligned quartz crystals in this very thin slice of the gemstone fire agate. The horizontal field of view is approximately 3mm. © Nathan Renfro

Polarized light shows vibrant birefringent colors from optically misaligned quartz crystals in this very thin slice of the gemstone fire agate. The horizontal field of view is approximately 3mm. © Nathan Renfro

Nathan Renfro is a photographer best known for his photomicrography of gemstones. A native of a small mining town in western North Carolina, Renfro received a BA in Geology at Appalachian State University in 2006. From there, he went to the Gemological Institute of America and got his Graduate Gemologist diploma in 2007. In 2014 Renfro earned his FGA (Fellow of the Gemmological Association) diploma from Great Britain’s Gem-A program.

 The surface of this diamond crystal shows triangular etch features known as “trigons.” This photograph was taken using differential interference contrast microscopy. The horizontal field of view is 1.2mm. © Nathan Renfro

The surface of this diamond crystal shows triangular etch features known as “trigons.” This photograph was taken using differential interference contrast microscopy. The horizontal field of view is 1.2mm. © Nathan Renfro

Renfro says that having a need to document what he was observing in the microscope is what led him down the road to photography.

“Because of what I had to learn about photography, I soon realized that I quite like photography in general, and often take the camera off of my microscope for landscape and street photography,” says Renfro. “By improving my photography skills outside of microscopic images, I think it helped develop my photography skills within the microscopic realm.”

 This image shows the geometric etch pattern in a synthetic rock crystal quartz. Blue, red and yellow filters were carefully placed below to highlight the trigonal symmetry of the crystal and provide vibrant contrast. The horizontal field of view is 5.43mm. © Nathan Renfro

This image shows the geometric etch pattern in a synthetic rock crystal quartz. Blue, red and yellow filters were carefully placed below to highlight the trigonal symmetry of the crystal and provide vibrant contrast. The horizontal field of view is 5.43mm. © Nathan Renfro

Though he says a geology degree isn’t necessarily required to be a photomicrographer of gemstones, it is helpful in order to really understand what he is looking at in the microscope.

“I think it is also important to be able to explain to the viewer what they are looking at and how the image represents a real tangible natural object even though it may look very abstract without that information,” Renfro says.

 Thin film interference colors are seen in this partially-healed cleavage crack in a natural topaz crystal. This image was taken using oblique reflected fiber optic illumination.  The horizontal field of view is 2.13mm. © Nathan Renfro

Thin film interference colors are seen in this partially-healed cleavage crack in a natural topaz crystal. This image was taken using oblique reflected fiber optic illumination.  The horizontal field of view is 2.13mm. © Nathan Renfro

His Process:

“The first step begins with finding a suitable subject. Not all gemstones will be particularly photogenic. Others will have great potential, but may have other limitations, like orientation issues, or scratches and blemishes on the gem's surface, which can really ruin what would otherwise be a great image. Sometimes I have to re-polish or re-cut gemstones in order to get the best photo from the stone that I think is possible.

 This image shows the surface of a beryl crystal. This was photographed using Differential Interference Contrast microscopy. The horizontal field of view is 1.2mm across. © Nathan Renfro

This image shows the surface of a beryl crystal. This was photographed using Differential Interference Contrast microscopy. The horizontal field of view is 1.2mm across. © Nathan Renfro

"Once I have found a suitable stone and have done any repolishing or sample preparation necessary, it usually doesn't take very long to find the right lighting and capture the image. I use lots of combination lighting including diffused transmitted light, oblique fiber optic illumination, polarized light, and sometimes colored filters to get the look that I want. After I capture the images, they typically look a bit flatter and duller than what I observed in the microscope, so I usually process the files in Lightroom to make them look as close to what I originally observed as I can.”

 A natural heart shape etch feature was present on the surface of a beryl crystal. This image was taken using differential interference contrast microscopy. The horizontal field of view is 1.1mm. © Nathan Renfro

A natural heart shape etch feature was present on the surface of a beryl crystal. This image was taken using differential interference contrast microscopy. The horizontal field of view is 1.1mm. © Nathan Renfro

Renfro compares looking at gemstones in the microscope to “being an explorer that has entered uncharted territory.”

“In all probability, when I look at a gem in the microscope, I may be the first person to ever really see what is inside,” Renfro says. “I get to discover new alien worlds filled with some of the most amazing examples of natural beauty. Having a camera on my microscope just lets me bring others along the journey....and I get to do this without having to get on a plane, drive hours in a car, hike lots of miles. I just get to sit in my office and go on an expedition whenever I want, for as long as I want.”

 Numerous crystals of the mineral tourmaline are trapped in their rock crystal quartz host. Vibrant birefringent colors are seen using polarized light. The vertical field of view is 4.32mm. © Nathan Renfro

Numerous crystals of the mineral tourmaline are trapped in their rock crystal quartz host. Vibrant birefringent colors are seen using polarized light. The vertical field of view is 4.32mm. © Nathan Renfro

 A large crack in an aquamarine shows vibrant thin film interference when using oblique fiber optic illumination. The horizontal field of view is 3.25mm. © Nathan Renfro

A large crack in an aquamarine shows vibrant thin film interference when using oblique fiber optic illumination. The horizontal field of view is 3.25mm. © Nathan Renfro

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