In Fall, 1971, I took the University of Wisconsin ­p; Madison Medical School histology course (Anatomy 710 "Cells and Tissues"), lecture and lab. Medical school courses had multiple instructors, and one was Allen W. Clark. Later, he agreed to teach me transmission electron microscopy, (a lot of work for him and for me!). I wanted to find out if the visual synapses of two non-phototactic Drosophila mutants (tan and ebony) were missing visual synapses. I could tell a volume of stories, but I will only tell three.

Allen had done electron microscopy (for instance, Clark et al. The ventral photoreceptor cells of Limulus I The microanatomy, J Gen Physiol, 1969, 54, 289-309). When I worked with him, he was doing electrophysiology on the effect of black widow spider venom on the neuromuscular junction. When I sat down at the dissecting microscope to prepare Drosophila heads, there were the spiders with their hour-glass abdomens, and I hoped they were dead and their venom glands removed.

Eventually, I obtained sections in an area that I thought might have visual synapses, but the most conspicuous structures were unknown to me (and to Allen [and to the departmental chair David Slaughterbach]). I eventually read some papers and found that these structures were called capitate projections (Trujillo-Cenoz, 1965, J Ultrastructure Res. 13, 1-33). My many electron microscope studies, described below, include an entire paper devoted to capitate projections (Stark and Carlson, Cell and Tissue Research,1986, 246, 481-486). The work with Allen Clark was done while I was a graduate student, and, while I was a graduate student, I met Stanley D. Carlson who would be central to my later studies (see "memoirs" in this link).

While I was an Assistant Professor at Hopkins, I published this work with Clark (Drosophila Information Service vol 50, pp. 105-106, 1973 ). It turned out that the synapses in tan and ebony were indistinguishable from wild type. The "article" was an un-refereed research note in a journal that most researchers did not take seriously. I had been told several times "Don't bother. Someone in Benzer's lab did that work." I eventually met that someone, Hansen, who had stopped publishing after getting tenure at Temple.

While I was an Assistant Professor at Hopkins, I thought it would be useful to develop EM techniques as part of my program of Research. Tina Ivanyshyn, an undergraduate, helped. I guess she was into equal rights since one time she did fixations on female Drosophila instead of males except that the mutant was maintained across from attached X, so the female was not mutant. Eventually, she got some nice pictures, and I included one in a grant proposal submitted in 20 copies, each micrograph carefully produced in the dark room and carefully taped to the page. A day after submission, the grants officer, Mr Proctor, called me in and showed me a mangled mailing label. "We sent in six that day and one was lost in the mail, but we won't know which one until we get the received notification for the five received and that can take a month. We've contacted all six grant officers, and they have agreed to give us an extension." Wouldn't you know it, it was mine, and I found out a day or two before my family vacation. Rush order in the dark room and in the production. I drove it from Baltimore to Washington and plopped it on the NSF desk in person. Eventually, my lost proposal (20? Copies) came back in a basket mixed with other items of mail.

EM did not get serious for me until 1981. I was at the University of Missouri, and, by that time, my friend Stan Carlson at Wisconsin had become an expert in several EM techniques. It is impossible to thank Stan Carlson as much as he deserves! I applied for and received a Faculty Development Award for $1,934. I went up there with the goal of looking at the synapses of the rdgB (retinal degeneration) mutant to see if the post-synaptic cells degenerated, a question left open after the studies with Bill Harris (1976, 1977). They do not degenerate (Stark and Carlson, 1982), so the trip was a success, and Stan pushed the work to publication so quickly. I was there Aug 2-29, stayed at the campus Y (very cheap) and then at the DeJonghs (friends from graduate school) when the Y needed space for students after Wisconsin's semester started. I learned so much! I was scared at all the tricks I would need to master. Oh, I could tell so many stories! But I will tell just a few:

Stan's wife, Che Chi, had done her PhD using EM on fly visual system. She came in one evening (after her work) and showed me how to section to the correct location, and that eventually became my methodology. I did some work at the high voltage electron microscope (HVEM). Founded by Hans Ris, this was an NIH Biotechnology Resource, so they were in the business of being hospitable (financially) to guests. My travel expenses for this August trip were complicated, and SM in the deans office helped with my travel expense voucher. Several years later, rumor had it, she got in trouble for some creative financial dealings.

Madison, WI is so nice in the summer! Ice cream at Babcock Hall (near where I worked, the Department of Dairy Husbandry in the Dairy State). Brats at the Memorial Union next to Lake Mendota with a rock band every evening. Jogging in Picnic Point or in the Arboretum. (but better bring plenty of blood since the mosquito is the state airline) Walking up and down State Street, pretty much like a boardwalk, after work. True I was away from my family, and my housing was dormitory style, but in later years at places right next to Lake Mendota. All my subsequent trips, mostly in the summer, were shorter. I used any excuse to get up there in a collaboration that lasted about 20 years. So, obviously I need to condense hundreds of stories to just a few.

Philippa Claude, at the Wisconsin Regional Primate Research Center, had a freeze fracture apparatus that Stan and I used in 1983 and again later. My friend from graduate school, Joe Kemnitz, worked at the Primate Center (and became Director years later [and had become my older son's Godfather years earlier]). We answered the question whether the diminished photoreceptors of the outer rhabdomeres absent (ora) mutant had rhodopsin (Stark and Carlson, 1983) [they do not]. I remember one amazing detail about a later trip. We were ready to work, but there was no liquid nitrogen that morning. For perhaps the only time in my career, I felt like Superman when it came to ordering, "OK, Badger welding is leaving a tank at the loading dock at 1 o'clock." I became an Affiliate Scientist at the Primate Center; they liked my contributions to their annual reports since I published. Eventually, Philippa had to get rid of the freeze-fracture apparatus, so she gave it to me. I went up for a day in January 1991 with my wife and my younger son to make arrangements, and the highlight was a Badger hockey match.

The HVEM was a way to make microscopy easy since we used thick sections rather than ultrathin sections, so my initial love affair seemed like cheating. One time, Missouri gave me about $240 for a plane ticket, and HVEM gave me about the same amount, so my (cheap) expenses were covered. (I had to fly the first few years since we had only one car, but later I drove.) The NIH Biotechnology Resource evolved (and they mothballed the HVEM), became IMR, Integrated Microscopy Resource, then LOCI (Laboratory for Optical and Computational Instrumentation). So many people helped me so much! Eventually, I tried all their tricks but focussed a lot on confocal microscopy at LOCI with a huge amount of help from Charles Thomas. By the mid-1980s, I did electron microscopy in my own lab with Randy Sapp as my grant-salaried technician.

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posted 7/25/16