Focused Ion Beam H-Bar Technique

Back in school, I learned how to use a Focused Ion Beam (FIB) workstation (taught by Bill Carmichael at MATC Madison). This interesting technology uses a Gallium source to create a beam capable of milling away at very small objects. This machine is used by technology companies such as Intel to aid in the creation of everything from computer chips and data storage devices to LCD displays and C-MOS digital camera detectors.

One technique commonly used by the industry for looking at and editing errors that occurred in the lithography is called the H-Bar technique. The H-bar technique produces an electron transparent cross-section (image top right click to enlarge) of an integrated circuit.
The microchip is polished to an approximate thickness of 20um and mounted to a grid (a 3mm circular piece of metal that can support a sample and is be placed into a Transmission Electron Microscope for examination (image left click to enlarge)). After putting the sample into the FIB, a small Tungsten strip is deposited to protect the circuits and then the sides of the microchip are milled away with the Gallium Ion beam. This results in an H-shaped cross-section of circuits, hence the name, "H-Bar." The sample can then be put into a TEM and the circuits imaged.
The above image was taken on a Hitachi H-800 TEM. The image below (click to enlarge) was taken with an FEI 610 Focused Ion Beam Workstation. Illustrations were made with AppleWorks 6.

Pollen Animation

If you've ever wondered about pollen, here's a good one for you. I found this interesting feature while examining a small grass flower in a Scanning Electron Microscope. Click on this link to see the animation. The pollen is sitting on the inside of the petal and as the image rotates, the object that comes in from the top-left and eventually covers up the pollen is the top side of the petal. I apologize if this takes a while to load on your computer. It is a rather large image file. I found that Firefox displays it smoother than Safari.
To produce the animation entailed taking 61 different images with the microscope. After taking each image, I tilted the sample 1 degree in the Z directions and reoriented in the X and Y directions in order to bring the area of interest back to the center. Then, additional focusing a stigmation correction was required in order to keep the image looking sharp. Once all off the data had been collected, the images were adjusted for brightness and contrast with Photoshop and the .gif was made with Image Ready.
These images were taken on a Hitachi 2700 SEM in the Madison Area Technical College Electron Microscopy Lab. The image on the right was colorized in Photoshop.

Armadillidiidae gonna eat you

Along the same lines as the guppy teeth, here are a couple of neat images I took of an Armadillidiidae (sometimes called a potato bug or pillbug).
I remember my friend trying to fry one of these guys with a magnifying lens when we were kids. The poor thing rolled up in it's little ball and a minute later we heard it sizzling. But I guess that's no worse than what this guy went though in order for me to be able to put him into the microscope. Sample processing for SEM of biological material consists of fixation with aldehydes and Osmium tetroxide, dehydration with a graded series of alcohol, something called critical point drying, and finally sputter coated with gold. Obviously the insect wasn't alive during all of this. I put him in the refrigerator beforehand. The cold puts them into a sleeping/hibernation mode. Likely he felt no pain.
These images were captured on a Hitachi 2700 SEM.

Guppy Eye

If you couldn't tell by now, I did a big project with guppy embryos when I was in school. Here's another image. Click to enlarge.
This is a 500 nm thick cross section of the eye of a guppy embryo. The lens (bottom part) of the eye was a bit deformed, but I think the rest of it survived the sample processing in tact. It was stained with Polychrome Blue and imaged on an optical light microscope. The interesting thing about this image is how it was constructed. I used a 40x lens and captured about 40 images. They were then manually stitched together in Photoshop (I was using CS1, when there was no option to do it automatically). Once completed, the image was 14,400 x 10,800 pixels (about 155 megapixels) and storage about a half of a gigabyte! I drastically reduced the size in order to publish it here, but I think it still looks pretty crisp. The original version could be zoomed in and viewed at high magnification.

D'na..d'na, d'na, d'na, d'na..wrEEEEEEEE! *jaws theme*

Did you know that guppies have teeth? I didn't 'till i put it in the SEM. This is another one from the guppy embryos back in school.

These images were taken on a Hitachi2700 SEM.

DLP Mirrors

Here's another entry of some images from when I was back in school. One day, my professor (Bill Carmichael at MATC Madison) brought in a DLP chip from a DLP projection television. The concept of how a DLP television works is quite unique. The chip is set up as an array of tiny little mirrors. Each one moves individually to project tiny bits of the image. With most DLPs, each mirror is responsible for one pixel.


Due to a mishap in the sample preparation of the chip, some of the mirrors fell off of their mechanical posts. This turned out to be a happy accident as it exposed the machinery so that more than just an array of mirrors could be seen. I captured these images on a Hitachi 2700 Scanning Electron Microscope. Click on each image to enlarge them.

A few pics

Let's add some fun pics. I've been in the EM field for a few years now. Here are a few images from back in school. All can be clicked to view them larger. More current work will follow.
This is a diatome. In the background are broken up ones. This was gathered from some diatomaceuos earth that we bought to keep slugs away from the veggies we were growing. The image was gathered with a Hitachi 2700 SEM and colorized in Photoshop.


This is some moss I found growing between two pieces of sidewalk. The image was gathered with a Hitachi 2700 SEM and colorized in Photoshop.


Here are a few images of some gills from a guppy embryo. Some were freeze fractured with liquid nitrogen to expose the contents. The images were gathered on a JEOL 840 SEM. One was colorized in Photoshop.