Microphotography - how to focus the digital camera through the microscope:

How to obtain sharp, in-focus photographs through the microscope using a low-cost digital camera.

The purpose of this paper is to help microscopists photograph, store, and use digital microphotographs using modern digital cameras and transmitted-light microscopes.

Focusing tips for digital photography at the microscope

Discussion focuses on selection of cameras 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 photograph at page very top is one of our earliest attempts at photographing Aspergillus sp. using lacto phenol cotton blue stain. Even a beginner can obtain very good microphotographs with just a little care. The photograph shown here is of one of our stereoscopic microscopes in use for making digital photographs.

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.]

PAAA 2005 Symposium, University of Tulsa, Tulsa Oklahoma - June 2-5, 2005 - updated 11/19/2023, 02/23/2009, 2015/10/25
Note: the latest version of this document can be found at InspectAPedia.com/Microscopy/Digital_Photography_by_Microscope.php
last update 20 August 2007 © Daniel Friedman All Rights Reserved

Hold the camera dead still against the eyepiece while shooting; the rim of the camera lens should press against your eyepiece sides, so you're not scratching anyone's lens. Or use a standard lens mount that replaces the eyepiece, or use a microscope with a trinocular head and mount your camera there.

• On our forensic microscope we mount the camera on the trinocular head and we add a Nikon MC-EU1 remote control which connects to the camera by a cable and permits us to take photos using a button on the desktop, avoiding touching the camera at all for least disturbance. However this device is not really necessary. This may be the only instance where the shutter delay on older digital cameras is an advantage. Simply press the shutter button and then remove your hands from the camera, giving it time to settle down before the shutter activates.
• If you're not getting a sharp photo the camera may be auto-focusing on a different part of the image.
• Be sure the object of interest is located in the center of the viewing area or use the camera's optional "manual focus" mode to select which area of the field is to be focused-upon.
• If you find the camera just can't get a clear photo no matter what, it might be that the image of interest lacks sufficient points of contrast (lines work best) for the camera to "see" what to focus upon.
• Try turning off the camera's auto-focus by setting it to focus at infinity. Then with the camera turned on, adjust the microscope stage up or down for the sharpest image shown on the camera LCD screen. You can improve your performance doing this by zooming in on the camera during focusing, then zooming out to get the image you want.
• For example shooting Periconia if you focus the microscope on the warty surface the edges of the spore will be blurred and it may take several tries at 100x to get a good shot. If you re-focus the microscope to show the sharp edges of the spore you'll see the camera hesitates much less before deciding it's got the image in focus.
• Try several shots, review on the camera screen, then load into your PC for really better examination.

Take some test photos through the microscope

Focus your microscope on your slide for best image Try shooting first through the eyepiece that has a reticule or other scale - you'll see the camera sometimes focuses on the scale rather than the image but at times, if the particle is close to the scale in depth this gives sharp results;

Try now shooting the same image through the other eyepiece - one with no scale mounted in it - forces camera to focus on the particle - which we put in the center of the field. Also we set this eyepiece on our microscope to "0" image adjustment since we think the camera would not prefer to have to focus through the correction we otherwise set for our own eye.

Test Images for evaluating image resolution:

you may already have images on file suitable for this purpose. we use slides of diatoms or radiolarians - particles which have very fine detail and sharp edges. One can also purchase slides specially prepared for this purpose (but they are expensive.) we tried using our stage micrometer but other images have finer detail.

...

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