Amino Acid Loto


© José Manuel Martínez MSc

Hi! I’m José Manuel Martínez, and recently, thanks to my image “Amino Acid Loto” I have been honored with the Premier Award and the Charles Foster Memorial Citation prizes in the BioImages 2022 awards by the BioCommunications Association. An additional perk of these awards is that I have been invited to talk about the technical and artistic aspects behind the image. So, this is the story.

I’m an electromechanical engineer with a MSc in production engineering. My first jobs were in drive systems design and finite element analysis. Not really related to microscopy, life sciences, or photography. But since 2006, I have worked in my own company, Quimica Tech, where I perform as a microscopy specialist. When people ask me what I do for a living, I usually answer: I play with microscopes and teach others to get the most out of their microscopes.

I took my first photograph with a microscope in 2012 when, in an emergency, and with only one week to learn everything about microscopy, I had to substitute an experienced microscopist that would be the speaker in a hands-on microscopy workshop. That first image was a cut of a convallaria in fluorescence, and it made such an impression on me that I started to bring random things under the microscope. One year later, I was playing with a confocal microscope, and people started to ask me to teach them how to use a microscope. This forced me to learn microscopy, and I’m thankful for that. In the following years, I continued taking things under the microscope and talking with regular users of microscopes. I chatted with molecular biologists, geologists, material scientists, virologists, quality engineers, high school teachers, and even art restorers. To all of them, I always asked: Do you have a sample you wouldn’t miss? Then I brought those samples to any microscope available; width field, confocal, or SEM (all of them usually borrowed microscopes) and then tried to get the best image possible. Most of those samples were burned by the lasers or just irreparably damaged. But in some cases, the owners were amazed by the photograph and wanted to use the image in their reports or papers. This way, I ended with 8 Terabytes of unedited micrographs, and around 2018, a friend told me, your images are amazing; why don’t you post them on Instagram? So, I opened the IG account of Quimica Tech (quimica_tech) and found other microscopists that fascinated me with their images, like the fluorescent snail of Håkan Kvarnström (aka micromundus photography), the perfect snowflakes of Michael Peres, the confocal images of Tagide de Carvalho, and the mesmerizing crystals of Karl Gaff (aka electron_micronaut). Lately, I have felt influenced by Marek Mis and his “minimalistic” way of preparing samples and his stunning images.

In Química Tech, part of my job is to challenge microscopes, cameras, and software; so I can explore how to advance microscopy techniques and solve problems that final users have. This has given me the ability to “forecast” how a sample would look under a specific microscope setting.

I regularly take pictures like the “Amino Acid Loto” in a combination of hobby and work. In my job, I run diverse types of samples in different preparation levels; I use amino acid crystals to explore different microscopes settings that I can later apply to technical applications. But I will not lie; sometimes, I drop my “paperwork” in the office to play with the microscope and try a new microscope’s setting just for fun. The Amino Acid Loto, was one of those drop-out moments where I just went to play with the microscope.

Many people ask me about “my creative process” to take micrographs, and most of them fill disappointed when I tell them I have no idea how I came up with the settings or sample preparation process to take that image. For me, explaining microscopy is easy; if you understand the concepts of multiplication and division, I can teach you how to use a microscope. But the creative process is something that I don’t even know what it is, and I can’t explain it.

Capturing images of crystals under the microscopes sounds easy at the beginning. One method is you first prepare a saturated solution of a chemical, which could be amino acids like in my case. Then put some drops of the solution on a microscope slide, smear it on the slide, let it dry, observe the crystal in a microscope with crossed polarizer’s filters, and Voilà! You have your crystals and images. In reality, you have to solve countless challenges and technical issues or at least be aware of them if you want to have a nice image. The “easy” factors to control are the concentration of your solution, the chemicals' purity, the temperature of the solution, and the dissolvent’s pH. Then are the factors that are hard to control, like the crystal’s layer thickness, the slide's temperature, and the slide's cleanliness. And then, are the factors that you must just accept and live with them, like room temperature, atmospheric pressure, the slide temperature, and the list keeps going.

For the image of “Amino Acid Loto” the greatest difficulty was maximizing the slide's cleanliness; the tiniest particle of dust on the slide could start nucleation. This will increase the number of crystals and reduce their size. In this case, I wanted a single nucleation point and one single big crystal.

For making the slide with the crystals, I prepared a solution of beta-alanine and glutamine in distilled water. Since I appealed to chaos, I didn’t measure how much of each component I used to make the solution, but I would advise taking notes and writing down how you prepare your sample so you can reproduce the results as pleased. Once I had the solution, I filled a syringe with the saturated solution; this way, I could dispense tiny drops on a microscope slide. Then I put about 10uL on a slide and heated it with an alcohol lamp just until the solution on the slide started to boil; immediately, I removed the slide from the heat and smeared the solution using another slide (just like blood smear in hematology samples). This way, a more or less homogenous layer of the amino acid mix will cover most of the slide, and within 10 to 20 seconds, this layer will start to crystalize in the most amazing ways. The forms that the crystals will take will depend, among an endless of factors, on how many solutions you put on the slide, how much you heat the slide, how fast-slow you smear the solution, how clean the slide is, how fast the slide cools down, and so on.

As I mentioned, I have 8 Terabytes of images taken with microscopes, but not all are worthy of publication or posting. When I judge an image or video taken with microscopes, first, I always ask: What is the purpose or objective of this image? Is it for scientific, educational, communication, quality control, or artistic purposes? If it is for scientific or quality control purposes, a great image must look nice but also provide information; for example, how big a cell is or which colors are present so minerals can be identified. For education and communication, an image should be eye-catching and invite the observer to want to know more about the sample. And when Art is the main reason for the picture, it should spark an emotion in the observer. As a microscopist, I always consider how difficult it could be to prepare the sample and how difficult it is to set the microscope and camera to take the image. For instance, images from a Transmitted Electron Microscope may look “noisy” and “granulated,” but when you understand all the work needed to prepare the sample, the scale, and all technical aspect, then the odd beauty of the image arise.

Finally, if you want to be more creative in your micro-imaging, I recommend not forgetting the basics; learn the microscopy theory (it is easier as many people think it is), know the optical properties of your sample, and then don’t be afraid to try something new. “Play” as much as you can with your microscope, and try to use as many contrast techniques as possible; it doesn’t matter if the techniques are not adequate for your sample. Just do not damage the microscope). You may burn a couple of samples before you find the right settings. My success rate is 10 to 1; I destroy, burn, or irreparably damage 10 samples for each “nice” micrograph.