Photographing Through a Microscope
Photomicrography - Inner Space

by Dr. Robert Berdan
April 24, 2017

Array of photographs taken through a light microscope with specimens including: wool, caffeine, bone, muscle, potato starch grains and a wide variety of crystals. I have never seen more beautiful colours anywhere.

Photographing through a light microscope is easier then ever before with Digital cameras and even cell phone adaptors you can attach. Light microscopes are not exactly common, but a quick search on E-bay or Kijjii will reveal numerous used microscopes. In this article I will share some pictures I have taken through my microscope. I started taking pictures through the microscope over 45 years ago and this lead me to a career first in medical research and later to full time nature photography. Of course you don't have to take pictures with the microscope, but rather simply enjoy exploring "Inner space" and the microscopic world that is unimaginably interesting. You will discover alien creatures in pond water and intensely coloured patterns in crystals and other biological specimens. Before you buy a microscope it helps to know someone who can guide you. It's the same thing if you are buying a telescope, if you don't know what to look for you may not be satisfied with your purchase. If you are willing to learn more about the biology of micro-organisms that will also be helpful.

Left is a professional stereo microscope and the right is a professional light microscope both with cameras attached. Most cameras can be attached using an extension tube which can be attached with gaffer tape or black electrician tape if you don't have the photomicro tube. The tubes to attach your camera to the microscope can be purchased on the web (see my links section).

A good used starter microscope can be had for about $50 and up. I would avoid Toy microscopes, I had one and it was OK to get started, but their not good enough to take pictures. There are lots of special techniques that you can use on a microscope like dark-field, phase contrast, polarizing filters etc that can enhance the appearance of the specimens so there is a bit of a learning curve. At the end of the article I provide some links to additional resources. The best way to learn is to get some books or have someone knowledgeable show you. I am available for instruction for those living in Alberta. For students under the 16 I will provide a one hour free lesson (with a parent in attendance) to anyone interested to help you get started. I love photomicrography and would be happy to help promote it. I hope some of my pictures in this article inspire some of you to give photomicrography a try.

Caffeine crystals 400X

There are many types of light microscopes, some are inverted for viewing cell cultures or specimens in petri dishes. There are stereo scopes which are the easiest to use and can be used to view specimens like leaves, rocks, flowers with no preparation but are limited to a magnification of about 50X. The regular light microscope can magnify specimens up to about 1000X. Light microscopes can be simple or have special lighting (polarized light, dark-field, florescence, differential interference contrast etc) with some of these techniques costing thousands of dollars or more to add on. The advantage of some of these techniques is that they can reveal transparent cells, or microorganisms as three dimensional subjects or make the the invisible - visible! The easiest technique is dark-field and it can be simulated by putting a dime on top of the a filter over the light source to provide a cone of light.

My foray in photomicrography began as a teenager, I am shown above in my garage laboratory with my microscope on the left. On the walls I have pictures of mosquito larva, rotifers, copepods, potato starch grains etc.

Above a photograph of a Dog Flea. Found on some pets they are easy to prepare by flattening them and mounting them on a glass microscope slide 50X. Small insects like mosquito can also be easily prepared for view, butterfly wings will reveal interesting scales and there is so much more.

While I was Heritage research scholar at the University of Alberta, I used a very sophisticated inverted light microscope to study nerve cells in culture. This microscope had a video camera attached for time-lapse photography and micro manipulators to which I attached microelectrodes to record electrical signals form the nerve cells (shown below with my wife posing in front).

Single brain cell from a fresh water snail. The neuron extends small processes with feet or fingers called a growth cones. Phase contrast microscopy. 400X

I enjoy photographing things through the microscope because I believe its a strange world that only a few individuals ever get to see - its like another dimension or inner space.

Above is a common mosquito sticking its proboscis into the skin of my father (macro-photograph about 10X). At the top left inset is a microscope view of the proboscis about 100X.

Above is a flat-worm found commonly in ponds usually attached to the underside of water-plants. Note the 2 eyes in the top left (black spots). This organism is unusual in that if it is cut in half each half will regenerate and it is used to study cellular regeneration. I am using an expensive technique to view this subject with a 3D relief called Nomarksi differential interference microscopy or DIC. 100X

Above is a single strand of a filamentous algae called Spirogyra. The chlorophyll is wrapped in a helix and in the center cell you can see the cell's nucleus. This plant is common in most fresh water ponds are streams and looks like green horse hair. Differential interference contrast 400X.

Freshwater snail embryos viewed with Differential interference contrast microscopy 50X. The eggs can be collected and one can watch the embryos develop from a single cell into a snail over about two weeks.

Pond water reveals thousands of different organisms. Above is a ciliated protozoan that appears to have been feeding on smaller green algae. Cilia are small hairs around the outside that propel these organisms. 200X. Phase contrast microscopy makes these transparent animals easier to see and study.

Many micro-organisms are attached to water plants. Above is a single cell Vorticella which is attached to the plant by a stalk that can retract quickly and form a coil. This protozoan filters and feeds on bacteria in the water. 600X Phase contrast.

There are hundreds of different types of algae found in ponds, lakes, streams, bird baths or free standing pools of water. Shown above are some algae strands using a technique called Dark-field microscopy which make the filaments appear to glow. 400X

Above is a picture of fresh water diatoms, these single celled animals construct complex silica (glass) shells around them to protect them and in the process form beautiful shapes. I took this photograph many years ago and it is one of my best sellers and is shown on the cover of a biology book in the inset. 400X Dark-field microscopy.

Radiolarians are microscope organisms that live in the ocean and like Diatoms from intricate silica shells. Dark-field microscopy. 400X.

Above area pair of rotifers, organisms found in pond water that have two rotating wheels on the head end that they use to feed. Around them there are three Euglena which contain chlorophyll to make their own food, or they can also eat bacteria - so are both plant and animal as they can move about. 400X phase contrast microscopy.

Of course light microscopes play an important role id identifying parasitic diseases and are used to to examine blood samples regularly. What you may not know is that it is possible to identify whether a person is male or female by looking at their blood samples a discovery made by a Canadian Dr. Murray Barr at the University of Western in London, Ontario. Some white blood cells called Neutrophils show an inactive X chromosome (women have 2 X chromosomes), so one is inactivated and it can be seen as extra large knob now called a Barr body (see below).

Blood sample from a female, in the center is a Neutrophil (white blood cell) that has been stained to show the Barr body (inactive X chromosome) that is absent in male blood samples since males only have one X chromosome. 600X Bright field microscopy.

Above is a human grey hair (one of mine) viewed in a light microscope about 400X

Crystals in Polarized light and using a Wave plate can take on incredible colours. Subjects like human hair, starch grains, wood, wool fibers or crystals of caffeine, vitamin C and a variety of chemicals can take on an abstract appearance. Polarized light can be had by adding a polarizing filter to the light source and another one in the eyepiece. This will cause subjects with birefringence (molecules organized in a crystal like fashion e.g. human hair) to appear bright. When using a wave plated (can make one by attaching Scotch tape to a glass slide) this can enhance and add colour to the subject. I will references below for those interested in reading more about this technique.

Wolf hair viewed using Polarized light and a wave plate. About 400X

Using polarized light and a wave plate can make for some very beautiful images of crystals.

 

Potato starch grains in Polarized light with a wave plate. To prepare this specimen I simply took a small piece of potato and squashed it onto a microscope slide. 200X

Caffeine crystals form a wide variety of crystal patterns. I create the crystals by dissolving pure caffeine in water then drying them on a microscope slide. A hot plate can speed up the process. The crystals are viewed in polarized light using a quarter or half wave plate to enhance the colour. Using this technique reveals a wide variety of shapes and patterns from different chemicals including vitamins, medicine in pills etc. 400X

citric acid crystals 400X

Caffeine crystals 400X

Vitamin C Crystals 400X

Caffeine crystals 600X

Caffeine crystals - Polarized light 400X

Caffeine crystals - Polarized Light 400X

Caffeine crystals 400X

Caffeine is one of my favourite substances to use to Create crystals because it offer such a wide variety of patterns and colours. To a scientist the colours can reveal the thickness, refractive index and the orientation the molecules are taking when the form the crystals. I have seen similar pictures from beer crystals through the microscope.

White pine wood - polarized light and wave plate 400X

White Pine Polarized light and wave plate 400X

White pine section - polarized light only 600X

Lanthanum crystals under Polarized light 400X

Lathanum crystals 400X

Magnesium sulphate crystals in Polarized light 400X

Ascorbic Acid Crystals (Vitamin C) Polarized light 400 X

Of course static pictures don't convey how interesting it can be to look a pond samples where the creatures zip around, divide and feed on each other. Making movies through the microscope now with DSLR cameras that have a movie feature is easier than ever. The main thing you will need is a very light source since the exposure must be 1\30 of second to get 30 frames per second. I use an old slide projector which I point at the mirror under the microscope to reflect light onto the specimen. Below is a short movie I made and uploaded to You-tube for fun, but it will give you an idea of how some of these alien creatures move about.

Short video showing samples of life living in a pond as viewed with my microscope. The movie was made with a Canon 5D attached to my Olympus E- microscope. I used phase contrast so that the biological specimens are more easily visible.

Exhibit of my photomicrographs at the Calgary Science Center (photo by Rinus Borgsteede)

My home microscopy laboratory, with stereo microscope, light microscope with Phase contrast and polarization, laptop, centrifuge, scales, vortex mixer, light box etc most items available from Amazon.ca. "Geeks paradise".

Portrait or R. Berdan - still a Science Geek - by Dr. Sharif Galal with graphics from many of Robert's Science Publications - picture taken May 18, 2017.

To learn more start with some library books on microscopy and visit the web sites below. Many companies like Olympus and Nikon provide a wide variety of resources to help you take pictures and learn more about microscopes. I hope you enjoyed these images, some have the appearance of abstract art. I will put an article together in the future on Stereo microscope photography. RB

Links where you can learn more about Microscopy & Photomicrography

Digital Photography through a microscope
Photomicrography as an Art
Digital Photomicrography
How to buy the right microscope
Life in a Drop of Water - You-tube Video
Digital Camera adaptors for your Microscope
Photographing Micro-organisms
Nikon Small world Photo contest - Galleries
Olympus Photomicrography Contest Gallery - Amazing pictures
Smart Phone adaptors for Digital Photomicrography
Home Made Smart Phone Adapter for Photomicrography
Charles Krebs Photomicrography
Olympus Differential Interference Contrast Microscopy
Carolina Protozoa and Invertebrates Manual - PDF
Olympus Polarized Light microscopy image gallery
Digicamcontrol software for connecting you Nikon or Canon camera to a laptop - Free!
Diatoms Ireland - lots of great information about preparing Diatoms for microscopy
Diatom Arranging by Steve Beats - PDF
Diatom arranging - Smithsonian
Olympus Polarized Light microscopy resources
Normarksi DIC - how it works PDF
Phase Contrast vs Normarski DIC - Nikon
Super-resolution light microscopy- recent developments - Wikipedia
Wikipedia - Waveplates - explains what they are and how they work.

The Amateur Scientist, December 1977 Studying polarized light with quarter-wave and half-wave plates of one's own making December 1, 1977 — Jearl Walker. This article got me started using Wave plates for photo-micrography.

Check out your local library and other web sites if you are interested in learning more about taking pictures with a microscope. Life is truly amazing, one only has to look more closely to appreciate its beauty.

Authors Biography & Contact Information

Robert Berdan is a professional nature photographer living in Calgary, AB specializing in nature, wildlife and science photography. Robert offers photo guiding and private instruction in all aspects of nature photography and Adobe Photoshop training.

Email at: rberdan@scienceandart.org
Web site: www.canadiannaturephotographer.com
Phone: MST 9am -7 pm (403) 247-2457.

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