2. 1 DETECTING INSECT HEARTBEATS by Shawn Carlson, August 1996There's a lot going on down among the microns. What we perceive as a rigid surface squashes easily under a finger’s gentle pressure when viewed from. a distance of a millionth of a meter. Increasing the temperature sends objects at that scale into even more violent upheavals. Biological processes reshape many living things on this scale. For ex- ample, every beat of an insect’s dorsal vessel— essentially, its heart—flexes its abdomen by a few microns. Now, thanks to John R. B. Lighton, a biologist at the University of Nevada, these tiny movements can be readily detected. (Lighton is not only a world-renowned physiologist but also a kindred spirit to amateur scientists everywhere, always striving to find the most direct and least expensive solution to vexing experimental challenges.) He re- alized that by detecting the microscopic flexings of an insect’s body, he could in effect put a tiny stetho- scope on the creature. Lighton’s ingenious method allows experimenters to embark on a fantastic voy-age into the microscopic universe. Now anyone can detect movements as small as half a micron—about the wavelength of visible light—for less than $40. Lighton senses micromotions by using mi- nuscule magnets that he attaches to the moving objects. He then relies on a special sensor that picks up the variations in the magnetic field caused by the shifting magnet. The sensitivity of Lighton’s detector depends on the fact that all magnets are dipolar; they havea north pole on one end and a south pole on the other. These poles would cancel each other perfectly if they weren't separated by the length of the magnet. This self-cancellation quality makes the strength of amagnetic field fall quite fast over space. Tripling the range to the magnet weakens the field by a factor of 27—the cube of the distance. The size of the magnet sets the scale by which this falloff can be quantified. The closer together the magnetic poles are (that is, the smaller the magnet), the more rapidly the magnetic field changes over distance.That in turn produces a larger signal for a micron- size shift. It’s easy to get micromagnets. You can buy so- called rare-earth magnets from Radio Shack (part number 64-1895), which sell for less than $2 a pair. They are tiny disks about 0.48 centimeter in diameter and 0.16 centimeter high («by Ys inch). At the surface, the magnetic field, which is oriented perpendicular to the flat part of the disk, is about 20,000 times that of the earth. Even these tiny magnets are too big for an insect stethoscope, so crush one with a pair of pliers. Made from a brittle ceramic, they will shatter into little shards. You need to make sure, though, that you know the direction in which the magnetic field of these shards points. Using nonmagnetic tweezers, position a fragment ona piece of wax paper. Placing the second magnet underneath the paper forces the fragment to align with the bigger field. Then dab a dollop of paint or five-minute epoxy over the mag- netic speck. Once it sets, the magnetic fragmentwill easily peel off the paper. Make at least 10 of these magnetic chunks, all slightly different in size, to see which one works best for your application [see Figure 21.1). Lym Figure 21.1 An insect stethoscope relies on a chip called a Hall effect transducer (HET), which is held to within a centimeter of the subject by a 12-centimeter-long plastic arm. The device recorded the “heartbeats” of a cockroach nymph (graph). The heart briefly paused after the 70-second mark because the experimenter dis- tracted the nymph with a hand wave. A Hall effect transducer (HET) senses the changes in a magnetic field. A modern-day sili-con miracle, it is small, extremely sensitive, and easy to use. Lighton recommends model $894A1F from Honeywell Micro Switch in Freeport, Ill.; call 800-537-6945 for a distributor. A bargain at less than $20, this device changes its output by 25 millivolts for each one-gauss shift in a magnetic field. (But be advised, Honeywell’s minimum order on this part is five units. However, the Society for Amateur Scientists is making this part available in single quantities. Contact them at SAS, 5600 Post Road, #114-341, East Greenwich, RI 02818; 401-823-7800 or www.sas.org.) Secure your HET no more than one centimeter away from your subject. For instance, if you are monitoring insects, you can epoxy the HET toa rigid piece of plastic and hold it above the subject with a device called Helping Hands, a soldering aid sold by Radio Shack. AHET records all magnetic fields, including the earth's. This indiscriminateness means that the de- tector will always produce a large constant voltagesignal (created by the earth and the magnet). On top of this voltage constant will be the small chang- ing signal created by the magnet’s motion. Most op-amps aren’t very good at detecting asmall changing signal that rides atop a large constant one. When faced with such a challenge many experienced experimenters rely on special devices called instrumentation amplifiers. Like op- amps, instrumentation amplifiers are available as. inexpensive, integrated circuits. Entry-level devices cost about $5; the Cadillacs of these chips sell for about $20. The AD524 from Analog Devices in Norwood, Massachusetts, works well; to order, call 800-262-5643, extension 3. You can also construct an instrumentation amplifier from three type 741 op-amps, but you'll probably have fewer headaches if you go with a prefab unit. If you ever want to use this technique to monitor temperature or some other signal that varies slowly, use Lighton’s slowly varying signal rendi- tion of the circuit. For flexing insect abdomens andother activities that change significantly over 30 seconds or so, use the quickly varying signal circuit. The circuit employs a clever technique that should be in every amateurs (and professional's) tool kit The method splits in two the voltage from the HET. One signal goes into the amplifier’s positive input. The other goes into a low-pass filter that only passes signals that oscillate slower than about one cycle every 30 seconds. Because an insects heart contracts in much less time, the filter strips out that signal and passes the large constant voltage (the DC offset). This filtered voltage is then fed into the instrumentation amplifier’s negative input. An instrumentation amplifier boosts the difference between its two inputs, so the troublesome offset voltage is automatically subtracted, leaving only the coveted signal /see Figure 21.2].Figure 21.2 Rate of motion dictates the necessary circuitry. Ifthe signal changes much over about 30 seconds, choose the quickly varying circuit, For more leisurely signals, use the slowly varying circuit. The instrumentation amplifier can be constructed from three operational amplifiers. A filter circuit and a power supply complete the system. Signal wires can introduce extraneous signals. They act like antennae, picking up electromagnetic energy, such as emanations from 60-cycle power lines, and then dumping it directly into your ampli- fier. To minimize the effect, keep the leads between the HET and the amplifier short. Additionally, you should use shielded wire. Lighton relies on three- core shielded cables. An electronics store may stock them, or you can make your own. Twist together three different colored wires, one each for the posi- tive, negative, and signal leads of the HET. Wrap the wires inside a layer of aluminum foil and connect the foil to the circuit's ground with a short wire. For protection, add a layer of duct or electrical tape around the foil. The filter circuit provides anotherbarrier to power-line noise. Finally, encase all your electronics inside a grounded metal project box. You can read the output with a digital voltmeter or, better yet, use an analog-to-digital converter to record the data into a computer. For the most up-to- date suggestions, check out the Society for Amateur Scientists’ web site at www.sas.org. Use shielded coaxial cable for the output connections, to prevent the HET from detecting the signal. Lighton obtained some remarkable results after he superglued a whole rare-earth magnet to the abdomen of a Blaberus discoidalis nymph, a relative of the American cockroach. With the instrumenta- tion amplifier’s gain set to 100, the signal caused by the contractions of the dorsal vessel—the insect’s heartbeats—is striking. After about 70 seconds of recording data, Lighton waved his hand in front of the nymph. The insect’s heart stopped beating for several seconds. According to Lighton, that hap- pened because the creature's nervous system maybe too limited both to maintain circulation and to attend to stimuli. Of course, any crawling by the insect will disrupt your results, so record data only when itis still. If the insect moves, it will generate a huge voltage signal that jumps well off the scale. In fact, Lighton reports that large signals occur whenever the insect opensits spiracles to breathe, about once every 5 to 30 minutes. By lowering the gain of your instru- mentation amplifier, you can also monitor insect respiration.