Digital Photography and the Microscope: How to take successful digital photographs through the microscope
MICROSCOPE DIGITAL PHOTOGRAPHY - CONTENTS: How to take photographs through the microscope - how to use a digital camera to take photographs through the microscope, Recommendations for digital cameras, adapters, photograph organizing software for microscope digital photography, Recommended camera adjustments and settings for use with a microscope
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Digital microphotography guide: The purpose of this paper is to help microscopists and other photographers take high quality digital photographs, store them for easy editing and retrieval, and use digital microphotographs
as an information or forensic database. We discuss using modern digital cameras and transmitted-light microscopes together.
This article discusses the selection of cameras that adapt well
to microscope eyepieces or trinocular heads, and on camera settings and procedures to obtain best quality photographs. We also discuss image resolution and size settings and make recommendations. Cameras used as examples in this paper
include the Nikon Coolpix series 990, 995, and 4500.
The techniques discussed in this paper work well with digital cameras and any type of microscope,
low power stereo zoom to high power forensic polarized light microscopes.]
Before specifying a camera choice and recommending camera settings let's review the purposes of digital microphotography of
particular interest to aerobiologists and other microscopists:
To develop a large reference library of photo images of known particles - so images will be mostly stored on a computer, in some data
base form. Images need to sharp, often with multiple exposures at different focuses.
For research: to exchange email images with
other professionals to obtain assistance in identification - similar to #1 above, but images by email should be
reduced in file size for convenience and transmission speed.
To print with professional reports - usually these are
limited in physical size. This is a distant third in importance in our practice.
Some Factors Involved in Obtaining and Using Good Photomicrographs
Start with a well prepared slide, choosing a mountant with good optics for the particles being examined.
Fungal spores or pollen grains may
benefit from hydration but not excessive hydration. Heavy use of colored stains risks obscures natural colors and
features needed for identification, though we may succumb and use fuchsin, lacto phenol, or even India ink for
occasional development of certain features.
After a well prepared and clean slide, be sure the microscope is properly set up for Koehler illumination, and be sure
the objective is clean.
Select a camera which has a very good lens - among the current crop of 3+megapixel cameras, this is perhaps more
important than a higher number of pixels. Lens resolution in macro mode and with the camera focused at infinity are
Post processing of images may improve the image color balance, sharpness, or contrast, but no amount of processing can
produce accurate image data if that information was not captured in the first place.
Even excellent photomicrographs are not useful if they cannot be located later for reference purposes. A good
photographic data base system is important.
The Digital Image Formation Chain determines image quality in digital photos through the microscope
Microscope resolution, including the quality, cleanliness, and adjustment of every lens in the light path. Before
starting the process be sure the microscope is clean and that K�Koehler illumination has been established.
Light source (color temperature, focus, etc.)
Adjustment of the condenser (N>1 oil-rated condenser lens for high magnifications such as the 100x
oil-immersion objective - don't forget to oil the top of an oil-rated condenser lens when using the oil objective)
Adjustment of the field diaphragm, condenser aperture, and where provided, objective aperture and eyepiece aperture)
Placement of the well-prepared and clean slide on the microscope stage
Choice of objective lens and use of oil-immersion (higher magnifications reduce the
demands on the camera's photo array of pixels)
Eyepiece and camera mount adjustment (and where provided,
calibration of the camera lens focal distance to match the eyepiece distance - or infinity)
Camera lens resolution
Pixel resolution of the camera used
Camera settings used for image capture
Camera's focusing or macro-focusing algorithm if it is not being used with lens set at infinity
Image data compression selection, ranging from zero (tiff) to low-loss, to high-loss compression
Post capture image processing
enhancement and cleanup, such as using the "digital camera" or "jpeg" cleanup functions
of image management software to remove noise, sharpen edges, adjust color balance, adjust contrast
dithering algorithm on compressed-image expansion
Level of image compression, e.g. JPEG, used to store final image - determines file size, maximum print resolution,
and limits of ability to crop and enlarge or "zoom" into the image.
Image display or printing capability, including effects of printer capability, dpi, dot
size, dot spacing, ink types, and choice of paper used to prepare the print - but remember, that contrary to the argument
of one expert microscopist who teaches that there is no reason to store large images, if you want to be able
to crop and enlarge or zoom into an image you need to have recorded the original with sufficient megapixels and resolution.
Visual resolution ability of the human eye
The limits of light to resolve particles are pertinent throughout these steps, at the microscope during
observation and making of the photograph in particular.
What this list means is that the quality of the end result of photomicrography is limited by the weakest link in the
image-formation chain. For example, no amount of camera pixels will provide a sharp image of an object which is not
resolved sharply in the microscope nor will a high-pixel camera produce a sharp image if the camera's lens is of
limited ability. Beware: not all high pixel cameras have equally sharp lenses, so the "resolution" in
megapixels can be misleading. High megapixels defines how many data points of image are being recorded. But if the lens
and other steps in the image formation chain are not producing a sharp image, high megapixels means you're recording
a lot of fuzzy data.
With these preliminaries, let's look at choosing a camera, selecting camera settings, using mounts, etc.
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References for Digital Photography Through the Microscope - Micro Photography
A Short Course in Nikon Coolpix 990 Photography, Dennis P. Curtain, http://www.shortcourses.com - RECOMMENDED
Resolution of Digital Photomicrographs from Scanned Film, Theodore M. Clark, Microscopy Today, Feb/Mar 2001
Light, Michael I. Sobel, University of Chicago Press, 1987, ISBN 0-226-76751-5
ThumbsPlus® image software, provides highly functional image database including basic editing, cropping, enlarging,
enhancement of photos, image organization, keywords and comments attached to each image, and search capability -
HIGHLY RECOMMENDED for PC platforms. Typically less than $100. Network support available.
® image editing software, provides sophisticated image editing, available for PC and Macintosh computer
platforms. Not recommended for reference library use - this is overkill for image editing and lacks good database
functions. Typically more than $400.
iPhoto® image software, provides easy user interface and limited function to organize and print photos for MAC computer
platforms - not recommended, limited function, very wasteful of disk space, lacks good database functions. Free
included with new Apple computers or operating systems.
Photo Explosion® image software - free from Microsoft with other OS purchases. Untried.
Picture Easy software from Kodak - free with some Kodak cameras, simple organizing and photo manipulation.
Nikon View® software - free from Nikon with purchase of Coolpix cameras, easy image import and simple image
manipulation and printing. Great for printing contact sheets of photos (which themselves are pretty useless).
 Thumbs Plus image data base software from Cerious software, www.cerious.com is a top choice low-cost option), and
will be searched-through for reference purposes
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Kansas State University, department of plant pathology, extension plant pathology web page on wheat rust fungus: see http://www.oznet.ksu.edu/path-ext/factSheets/Wheat/Wheat%20Leaf%20Rust.asp
"A Brief Guide to Mold, Moisture, and Your Home",
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US EPA - Mold Remediation in Schools and Commercial Building [Copy on file at /sickhouse/EPA_Mold_Remediation_in_Schools.pdf ] - US EPA
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"A Brief Guide to Mold, Moisture, and Your Home", U.S. Environmental Protection Agency US EPA - includes basic advice for building owners, occupants, and mold cleanup operations. See http://www.epa.gov/mold/moldguide.htm
"Disease Prevention in Home Vegetable Gardens,"
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Fifth Kingdom, Bryce Kendrick, ISBN13: 9781585100224, is available from the InspectAPedia online bookstore - we recommend the CD-ROM version of this book. This 3rd/edition is a compact but comprehensive encyclopedia of all things mycological. Every aspect of the fungi, from aflatoxin to zppspores, with an accessible blend of verve and wit. The 24 chapters are filled with up-to-date information of classification, yeast, lichens, spore dispersal, allergies, ecology, genetics, plant pathology, predatory fungi, biological control, mutualistic symbioses with animals and plants, fungi as food, food spoilage and mycotoxins.