Immunocytochemistry

Lately in the lab, I've been doing a lot of Immunocytochemistry (ICC). It's a time-consuming process that ends hours later with a few prepared microscope slides to show for your work. The whole point of ICC is to determine whether or not the cells you're working with have certain antigen or receptor. In order to do this, we use antibodies that target specific peptide or protein antigens in the cell.
To start the process, I'll fix the cells I'm going to be testing with a solution of paraformaldehyde. This basically stops everything that's going on in the cell at the moment it's fixed. Then, I'll wash the cells (we grow them on coverslips) twice with a glycine wash to remove the paraformaldehyde. Next, I'll use a permeabilizing solution on the membrane. Finally, it's time to put on the first, or primary antibody, which will go on the cells, in an extremely diluted form (antibodies are extremely expensive...like hundreds of dollars for a quantity that is 1 milliliter or less). This antibody will be left on for the amount of time the researcher chooses, then is rinsed off in preparation for the secondary antibody. When selecting a secondary antibody, it is important to choose one that works with the primary that you used. For example, if the primary antibody was created in a goat, I'd have to choose a secondary antibody that has anti goat IgGs. Attached to the secondary antibody is a fluorophore (compound that makes a molecule fluorescent). This allows us to see whether the antigens exist. After the secondary antibody sits for an hour, the coverslips are washed 3 more times, then mounted onto microscope slides and allowed to dry. Finally, they can be viewed. I use the microscope and camera to view the slides. If the edges of the cells are glowing, this means the cells do indeed have the antigens the primary antibody tested for.

more pictures from the saguaro fruit harvest

Catching a falling saguaro fruit...

A really big cactus...

It's harder than it looks!

A random picture of waiting for the bus after a successful Trader Joe's grocery shopping trip.

Walking back to the camp as dusk fell after harvesting saguaro fruit.

Saguaro Fruit Harvest

To harvest the saguaro fruit, one person uses a long pole made out of a saguaro rib (they're inside the cactus, kind of like strong sticks that are the skeleton of the saguaro) to knock the fruit to the ground while the other person holds the bucket and tries to catch the falling fruit.

Eating saguaro fruit! It tastes like a very mild strawberry with lots and lots of black seeds.

The saguaro fruits are the green knobby things on the cactus. Some of them are open and you can see the red, seedy fruit inside.

You can practically feel the heat just by looking at this picture...

The first couple fruits that we collected.
This week we went to the reservation of the Tohono O'odom out in the middle of the desert to learn about and participate in the saguaro fruit harvest. We learned that these people believe the saguaro to be their dead ancestors, and the harvest is a very special time. However, this is a tradition that is largely dying, because many of the people can't take the time off from work to spend more than a month camping in the desert to harvest the saguaro fruit.

Harvesting goes like this: one person uses a long saguaro rib to hit the fruit down from the cactus, while another person stands beneath with a bucket to catch the falling fruit. It was fun, but also hard work.

After harvesting, we cleaned the fruit, which entailed peeling off the outer green covering to reveal the bright red seedy fruit beneath. We then ate a traditional meal of beans, homemade tortillas, and cholla (it's a type of nasty cactus) buds (which tasted an awful lot like artichokes).

As dusk fell, we listened to stories about the traditions of the Tohono O-odom people and marveled at the millions of stars that twinkled brightly in the black sky.

A Violin!

Last week I bought a violin! I had decided not to bring my flute with me to Arizona, but lately had been feeling really empty without being able to play music. I decided to check out renting an instrument, but Lindsay from my lab told me about a shop that consigns guitars, ukeleles, violins, mandolins, and other instruments, so I decided to check it out after work one day. After plucking the strings of several violins, I found one with a sound I liked, and saw that it wouldn't cost much more to buy the violin than to rent an instrument for a summer. I rode away, holding my new violin case in one hand, while still trying to use my bike brakes. I got quite a few interesting looks riding down Speedway with a violin case hooked through a handlebar, but made it back mostly in one piece. I'm excited to get to play again!

Work, Food City, and Various Thoughts...

Sun going down behind the train tracks...

Food City is our favorite grocery store! It's the best place to buy super cheap Mexican food, just make sure you know how to speak Spanish, because your transaction with the cashier will probably not be in English.

My bike ride back to my apartment on Speedway...it's a pretty busy road, with a great mountain view.

Entrance to the Medical Research Building, where I work. I don't think I've ever actually seen anyone sitting at these tables... ;-) The past few weeks have mostly been a big blur. At work every day, I either take care of my cells, feeding or splitting them, do experiments (which is basically taking pictures of groups of beta cells with a microscope/computer at specific timepoints, with variables of glucose concentration and after adding different ligands or drugs used for insulinoma), or analyze the data from the experiements using DOS and Excel. Tomorrow I'm going to learn immunocytochemistry.

The highlight of our week is the trip to Food City on Friday nights. The grocery store is...more than a little sketchy. It's about a mile and a half walk from our apartment, in a strip mall with a "Factory 2 U", "Dollar General" (where we got our potentially deadly pan), and a "G Town". We all stock up on our tortillas (made fresh daily!), Mexican cheeses, avocados, limes, cilantro, and all the other specialties of Food City while practicing our Spanish (most of the signs in the store are in Spanish). On Saturday, we figured out the bus system, which is not fantastic, by any means, but it got us to the mall, a used bookstore, and CVS. On Sundays, I usually ride my bike to church (3 miles each way, in the hot, hot sun), then go swimming, and get in some relaxation time.