z-stacked Photomicroscopy

•  Z-stacked photomicroscopy using an Ash Technologies Ltd, Omni Core full HD 1080p digital microscope provides 'in-focus' images for detailed scientific publication. It can be tricky at first to acquire good clear images and certainly specimen quality plays a part in this. Too much surface debris on the insect and the lens will focus on that debris rather than on the insect surface, so try to use clean, freshly caught specimens where possible.

Step 1. Camera settings.

From the side menu, select the cog-wheel icon representing Settings.

The default tab is Camera, but if Camera is not current, then select it.

Select the 'Clog' button: Show Advanced Camera Settings

• Increase Aperture to 15.
• Leave Noise L at the default setting of 3.
• Increase Contrast to about 150 (depending on your subject material).

Select Apply Settings and then select Okay

• Then either select Auto Exposure, or adjust Gain and Iris to gain optimal visual clarity.
• It doesn’t really matter if Autofocus is selected or not, but select it.

Select  Okay

Step 2. Depth of field setting

From the side menu, select the icon with four radiating arrows, labelled Overview.

Centre the specimen in the crosshairs.

Click the Overview icon again.

Zoom in to full zoom.

Using the focus control on the camera stand, focus on the specimen.

Using Zoom out, return the view to a magnification that accommodates your required image.

Step 3. Extended Image Focus

From the side menu, select the stack-sheets icon representing Extended Image Focus. This is the menu in which you set up z-stacking.

Select Ignore Focus Limits.

Maximise Images and Quality levels to 12 and 10 respectively.

Using the focus control on the camera stand, focus to the lowest point you wish to have in focus.

Click Bottom.

Now click Top (do not touch the focus control on the camera stand).

Focus to the upper most point you wish to be in focus using the up and down arrows.

Click start.

Vibration

If the bench you work on suffers from any vibration, be careful to move away and be still while the Acquire Images track scrolls to 100%. Once that has happened and Align Images and/or Stack Images is tracking you can safely continue to work at the bench.

Once satisfied, Save your image.

Not what you wanted? Try adjusting the Aperture to a lower value and/or increasing the contrast.

Still stuck? Sent me a message.